International Journal of Biological Macromolecules 170 (2021) 602–621

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

journal homepage: http://www.elsevier.com/locate/ijbiomac

Review

Biodegradable microneedles fabricated with carbohydrates and proteins:

Revolutionary approach for transdermal drug delivery
Neha Dabholkar a,1, Srividya Gorantla a,1, Tejashree Waghule a, Vamshi Krishna Rapalli a, Avinash Kothuru b,
Sanket Goel b, Gautam Singhvi a,⁎
a
Industrial Research Laboratory, Department of Pharmacy, Birla Institute of Technology and Science (BITS), Pilani, Pilani Campus, Rajasthan, India
b
Department of Electrical and Electronics Engineering, Principal Investigator: MEMS, Microfluidics and Nanoelectronics Lab, Birla Institute of Technology and Science (BITS), Pilani, Hyderabad
Campus, Hyderabad, India

a r t i c l e i n f o a b s t r a c t

Article history: There has been a surge in the use of transdermal drug delivery systems (TDDS) for the past few years. The market
Received 1 October 2020 of TDDS is expected to reach USD 7.1 billion by 2023, from USD 5.7 billion in 2018, at a CAGR of 4.5%.
Received in revised form 22 December 2020 Microneedles (MNs) are a novel class of TDDS with advantages of reduced pain, low infection risk, ease of appli-
Accepted 23 December 2020
cation, controlled release of therapeutic agents, and enhanced bioavailability. Biodegradable MNs fabricated from
Available online 31 December 2020
natural polymers have become the center of attention among formulation scientists because of their recognized
Keywords:
biodegradability, biocompatibility, ease of fabrication, and sustainable character. In this review, we summarize
Biodegradable microneedles the various polysaccharides and polypeptide based biomaterials that are used to fabricate biodegradable MNs.
Pullulan Particular emphasis is given to cellulose and its derivatives, starch, and complex carbohydrate polymers such
Hyaluronic acid as alginates, chitosan, chondroitin sulfate, xanthan gum, pullulan, and hyaluronic acid. Additionally, novel
Silk fibroin biopolymers protein-based polymers such as zein, collagen, gelatin, fish scale and silk fibroin (polyamino acid) biopolymers
Transdermal drug delivery application in transdermal drug delivery have also been discussed. The current review will provide a unique per-
spective to the readers on the developments of biodegradable MNs composed of carbohydrates and protein poly-
mers with their clinical applications and patent status.
© 2020 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 603
2. Mechanism of drug delivery from MNs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 604
3. Types of microneedles and drug delivery approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 604
3.1. Solid microneedles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 604
3.2. Coated microneedles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 605
3.3. Dissolving microneedles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 606
3.4. Hollow microneedles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 606
3.5. Hydrogel forming microneedles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 607
3.6. Biodegradable microneedles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 607
4. Biodegradable microneedles: mechanism, natural polymers used for their fabrication, and applications. . . . . . . . . . . . . . . . . . . . . . 608
4.1. Polysaccharides or carbohydrate polymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 608
4.1.1. Alginate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 608
4.1.2. Cellulose and its derivatives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 609
4.1.3. Chitosan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 609
4.1.4. Chondroitin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 609
4.1.5. Hyaluronic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 609
4.1.6. Sugars . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 609

⁎ Corresponding author.
E-mail address: gautam.singhvi@pilani.bits-pilani.ac.in (G. Singhvi).
1
Authors contributed equally.

https://doi.org/10.1016/j.ijbiomac.2020.12.177
0141-8130/© 2020 Elsevier B.V. All rights reserved.
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

4.1.7. Xanthan gum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610

4.1.8. Pullulan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610
4.1.9. Bletilla striata . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610
4.2. Polypeptides or protein polymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610
4.2.1. Zein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610
4.2.2. Collagen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610
4.2.3. Gelatin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 610
4.2.4. Fish scale polymer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 613
4.2.5. Silk fibroin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 613
4.3. Polysaccharide and polypeptide composites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 613
5. Biocompatibility and biodegradation of the polysaccharide and polypeptide MNs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 614
6. Clinical applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 615
7. Patent status . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 615
8. Future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 616
9. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 617
Declaration of competing interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 617
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 617

1. Introduction compatibility, improved patient compliance, and termination of dose

Human skin is a protective barrier that only allows one-way chan-
nelization. It prevents the entry of external matter into the body. Thus,
the skin resists absorption of chemicals that come in contact with it;
making drug delivery through the skin cumbersome [1,2]. Since the ap-
proval of the first transdermal drug delivery system (TDDS), Transderm
SCOP (for travel-associated nausea and vomiting) in 1979 by the FDA,
there has been a surge in the use of TDDS [3]. The market of TDDS is ex-
pected to reach USD 7.1 billion by 2023, from USD 5.7 billion in 2018, at
a CAGR (Compound Annual Growth Rate) of 4.5% [4].
The term ‘transdermal delivery’ is used for the diffusion of a solute
through the different layers of the skin to reach the systemic circulation
and elicit a therapeutic effect. For a drug to be delivered through the
skin by passive transport, it must be lipophilic and have a molecular
weight <500 Da. This limits the transdermal delivery of drugs. Active
and passive approaches have been used to optimize TDDS. In the active
approach, drug delivery is enhanced using physical methods. In the pas-
sive approach, the drug or the formulation is optimized to improve skin
permeation. However, the passive approach cannot be used for drugs
having molecular weight >500 Da [3,5]. Transdermal delivery of drugs
offers certain advantages including avoidance of drug degradation in
the gastrointestinal tract (GIT), overcoming intra- and inter-subject var-
iability, retention of the drug at the therapeutic concentration for a
prolonged period, minimization of dosing frequency, enhanced
simply by removing the patch [5,6].
The most commonly used TDDS includes hypodermic needles and
transdermal patches. However, the needles are less accepted by patients
because of the pain associated with their use, and transdermal patches
may be limited to delivery of small molecules as they have to diffuse
through several layers before reaching the systemic circulation [7].
MNs combine the advantages and overcome the shortcomings of hypo-
dermic needles and transdermal patches. Fig. 1 depicts drug delivery by
various transdermal delivery systems. A comparative description be-
tween topical cream, transdermal patch, hypodermic needle, and
microneedle drug delivery systems is presented in Table 1 [7].
The use of microneedles (MNs) is a novel, third-generation approach
to optimize TDDS [8]. MN delivery system was first proposed as a US
patent by Gerstel and Place in 1971 [9,10]. The MNs should be fabricated
with suitable length and depth devoid of stimulating the underlying
dermis and blood vessels. Typically, MNs are 250–1000 μm in length,
50–250 μm in width, and 1–25 μm in diameter. When the MNs are ar-
ranged in arrays on a patch backing, it is known as a MN patch [11].
The common terminology and parameters used in describing the archi-
tecture of a typical MN patch are presented in Fig. 2(a) [12]. A MN setup
has four components: drug reservoir, micropump, microflow sensor,
and MNs. The drug reservoir stores and retains drug formulation.
Micropumps inject the drug formulation from the reservoir to the nee-
dle. The flow sensor monitors and regulates the drug flow through MNs

Fig. 1. Comparison of topical cream, hypodermic needle, microneedle patch and transdermal patch (Reprinted with permission from ref. [7], copyright © Elsevier).

603
N. Dabholkar, S. Gorantla, T. Waghule et al.
Table 1
Comparison between topical cream, transdermal patch, hypodermic needle, and microneedle drug delivery systems.
Topical cream Transdermal patch
Description Emulsion/emulgel/cream/ointments Adhesive patch to be placed on the Fine, hollow tube having a
skin.
Onset of action Slow Slow
Pain Painless Painless
Bioavailability Poor Insufficient
Patient compliance less Better
Self-administration Possible Possible
Mechanism of drug Permeation through skin pores. Drug has to cross stratum corneum Drug placed directly in the
deliver barrier, thus poor diffusion of large dermis.
molecules.
[8]. At a commercial level, MNs have been used for the delivery of small
and large drug molecules. MNs pierce the stratum corneum of the skin
and produce micron-sized pore channels, which act as a direct route
for drug delivery [11].
Chen et al. reported that MNs maintained the concentration of insu-
lin for a longer period compared to s.c. injection. The maximum change
in glucose level was obtained after 1 h of administration of the MNs and
s.c. injection. The blood sugar continued to decrease in the subcutane-
ous injection group, reaching its lowest level after 2 h. In contrast, in
the MN group, the lowest level was reached after 3 h [13]. MNs are
also superior to transdermal patches. In a study conducted by
Wermeling et al., the systemic administration of a hydrophilic drug, nal-
trexone, was demonstrated using MNs. Upon transdermal delivery of
naltrexone using the transdermal patch, no plasma levels were detected
even after 72 h of application. However, upon pretreatment with MNs,
steady-state plasma concentration was obtained within 2 h of applica-
tion and the concentration was retained for at least 48 h. Thus, MNs
can provide a rapid onset of action [14]. The advantages of MNs include
ease of administration of large molecules [15,16], minimally painful and
invasive, decreased microbial penetration compared to a hypodermic
needles [17], targeting of specific skin area [18], dose reduction,
accurate dosing [19], rapid drug delivery by coupling with electrically
controlled micropump [20,21], rapid onset of action and self-
administration. MNs have been used for the transdermal delivery of a
wide array of therapeutic agents including, vaccines [22–24], insulin
[25,26], hormones [27], enzymes [28], contraceptives [29], and anti-
cancer drugs [30,31].
Despite having these many advantages, MNs possess some limita-
tions. Solid MNs fabricated by silicon, glass, metal, ceramics are used
as first-generation MNs. These are used for transdermal drug delivery
due to its tremendous mechanical strength and biocompatibility. The
solid MNs fabricated by silicon, metal, ceramics can overcome the skin
barriers but exhibits limitations such as limited drug loading, skin
break up, and costly fabrication procedure. The material used to fabri-
cate MNs, affects its permeability, flexibility, and toughness. In recent
days, to overcome these limitations, polymers are the most widely
used materials for MNs fabrication [32]. The biomaterials obtained
from various plant and animal kingdoms such as polysaccharides and
protein-based biopolymers have attracted significant attention in the
realm of MNs research. This review provides a view on the types of
MNs and drug delivery approaches properties of polysaccharides and
protein-based biopolymers which make them suitable for MNs fabrica-
tion. Additionally, it gives the recent update on the applications of MNs
fabricated from cellulose and its derivatives, sugars, complex carbohy-
drates and protein biopolymers in drug delivery.
2. Mechanism of drug delivery from MNs
There are two routes of penetration of drug across the intact
skin; transepidermal route and transappendegeal route. In the
604
International Journal of Biological Macromolecules 170 (2021) 602–621
Hypodermic needle Microneedle
Micron size needles are aligned on the surface
sharp tip with small of a small patch.
opening at the end.
Faster Faster
Painful Painless
Sufficient Sufficient
less Better
Not possible possible
Bypass stratum corneum, and drug placed
directly into epidermis or dermis, resulting in
enhanced permeability.
transepidermal route, the molecules pass through the stratum
corneum; intracellularly through the corneocytes, and intercellularly
through the intercellular spaces. The intracellular route allows penetra-
tion of polar or hydrophilic solutes, whereas the intercellular route al-
lows penetration of non-polar or hydrophobic solutes. In the
transappendegeal route, the molecules pass across the hair follicles,
and via sweat glands [33–36]. Understanding of kinetics is necessary
to design a TDDS. Percutaneous absorption is an important step in trans-
dermal delivery. The penetration of molecules into various layers of skin
and their permeation into the systemic circulation is termed as percuta-
neous absorption. It is a step-wise process that involves (i) penetration,
(ii) partitioning from the stratum corneum into the epidermis, (iii) dif-
fusion into the upper dermis, (iv) permeation, and (v) absorption into
the systemic circulation [33].
The mechanism of drug delivery from MNs is based on mechanical
disruption of the skin and delivery of drugs within the epidermis.
Thus, the drug reaches the target site readily bypassing the stratum
corneum barrier. The active substance is dissolved/dispersed/entrapped
in the MNs. After application, the MNs either create micron-sized chan-
nels in the skin or completely degrade in the skin releasing the encapsu-
lated drug without any hazardous residue left behind [37]. The MNs
pierce the upper skin layers and enable the delivery of the drug directly
into the epidermis/upper dermis region which is otherwise not possible
by passive diffusion alone. As micron-sized channels are created, even
macromolecules like hormones, peptides, and immunobiologicals can
be delivered efficiently.
3. Types of microneedles and drug delivery approaches
Microneedle is a micron-sized needle with appropriate size ranges
i.e. 150–1500 μ (height), 50–250 μ (base width), and 1–25 μ (tip diam-
eter) [38,39]. It directly pierces the stratum corneum barrier without
stimulating nerves and damaging blood vessels. The above-mentioned
dimensions are mostly used for transdermal drug delivery because of
advantages such as improved drug bioavailability, safety, minimally in-
vasive, and proficient drug delivery. Fig. 2(b) shows the transdermal
and lymphatic drug delivery using microneedle [12]. Morphologically,
MNs can be classified into five types: solid, coated, dissolving, hollow,
and hydrogel-forming MNs [40]. Each type of MN has its own way of de-
livering drugs into the epidermis. Different types of microneedles with
their drug delivery methods are presented in Fig. 2(c) [12].
3.1. Solid microneedles
Solid MNs use the “poke-and- patch” approach. Solid MNs are made
from different types of metals like silicon, stainless steel and titanium.
They are mostly used for pretreatment of the skin to create micropores.
Upon subsequent application of a drug patch, the drug enters the skin
with decreased resistance [9,20,32,41]. In a study, super-short solid sil-
icon MNs (70–80 μm) were found to be better than sharp-tipped MNs in
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

Fig. 2. (a) Schematic highlighting common terminology and parameters used in describing the architecture of a typical microneedle patch. A microneedle patch consists of micron
projections which are commonly known as microneedles supported on backing layer (or baseplate). The tip of a microneedle (as shown in the inset) may be characterized by the apex
angle or radius of curvature which dictate the skin insertion capability. (b) Lymphotropic drug delivery using microneedles. Microneedles with typical lengths of 250–1000 μm would
penetrate the epidermis layer, reaching the dermis layer of the skin. The dermis is rich in blood and lymph vessels and the drug released in this layer will readily have exposure to
various types of immune cells. (c) Types of microneedles: Solid, hollow, coated, dissolving and hydrogel-forming microneedle (Reprinted with permission from ref. [12], copyright ©
Elsevier).

enhancing skin permeation. Also, they are retained in the skin for a lon-
ger period compared to sharp MNs [42]. Narayanan and co-workers fab-
ricated solid silicon MNs and coated them with gold to improve their
mechanical strength (Vickers hardness value: 3800 Hv) and biocompat-
ibility [43]. Solid MNs made of metals and silicon have been especially
used in gene therapy [44]. Dermaroller® is a commercially available
solid MN device that has been widely used in dermatology [45]. Thus,
the solid MNs could be a promising candidate for increasing drug avail-
ability in transdermal drug delivery systems.
3.2. Coated microneedles
Coated MNs use the “coat-and-poke” approach. The biomolecules or
medications are coated on the MNs [9,20,32,41]. Kapoor et al. developed
a coated solid microstructured transdermal system (sMTS, developed
by 3 M) for a potent peptide A and investigated the preclinical evalua-
tion using two different coating solutions. Approximately 250 μg of
the peptide could be coated on 1.27 cm2 patch containing 316 MNs
with precision. The absolute bioavailability for MNs coated with solu-
tion 1 and solution 2 was found to be 88% and 74%, respectively,
which was similar to s.c. bioavailability of 75%. The stability of the pep-
tide at room temperature was improved. Additionally, skin irritation
studies on porcine skin demonstrated minimal skin irritation [46]. Li
and co-workers coated MNs individually, instead of coating all MNs
with the same formulation. Thus, individually coated MNs delivered
therapeutic agents with different physicochemical characteristics. The
number of coating layers was modified to adjust the MN loading. It
was found that polyvinyl pyrrolidone (PVP) coating dissolved within

605
N. Dabholkar, S. Gorantla, T. Waghule et al.
30 s in phosphate buffer solution. They reported that PVP K15 dissolved
even faster compared to PVP K30 [47]. The only limitation of the coated
MNs is that these give rise to safety concerns due to the biohazardous
waste left behind after use.
3.3. Dissolving microneedles
Dissolving MNs use the “poke-and-release” approach. They are fab-
ricated by encapsulating the drug into water-soluble materials (polyvi-
nyl pyrrolidone, chondroitin sulfate), sugars (maltose, dextran), or
biodegradable polymers (polylactic acid, chitosan). Upon insertion of
MNs in the skin, either dissolution or polymer degradation takes
place, subsequently releasing the drug. Hence, the MN is not to be re-
moved after insertion. This provides a one-step application process con-
venient for patients [9,20,32,41]. The choice of MN material affects the
drug release from the MNs. MNs can be used to attain controlled release.
In a study, it was found that biodegradable MNs can provide controlled
release of the drug for hours to months [48]. The drug release rate and
duration can also be regulated by controlling the rate of degradation
of polymers in the case of biodegradable MNs [28,49].
Lahiji and co-workers developed tissue interlocking dissolving MNs
using rhodamine B as the model drug. Sharp tip, narrow neck, and wide
body of the MNs resulted in the tissue-interlocking property. The mini-
mum force required to penetrate the skin was less for the tissue-
interlocking MNs (0.1±0.07 N) compared to conventional MNs (0.19
±0.06 N) [50]. The elastic nature of the skin prevents dissolving MNs
from complete insertion and accurate drug delivery into the skin. To en-
counter this issue, a patchless, micro-pillar integrated dissolving MN
was fabricated. Polymethyl methacrylate was used to make the micro-
pillars and hyaluronic acid was used to fabricate the MNs. Interestingly,
the patchless MNs increased the skin penetration of the drug up to
97.78% ± 2.22%, compared with 44.44% ± 7.85% in the conventional
Fig. 3. Schematic illustration of transdermal delivery of insulin using starch/gelatin microneedles (MNs), which can rapidly dissolve in the skin to release encapsulated insulin (Reprinted
with permission from ref. [52], copyright © Elsevier).
606
International Journal of Biological Macromolecules 170 (2021) 602–621
dissolving MNs [51]. In short, compared to solid MNs, the dissolving
MNs are easy to prepare, biocompatible, provide high drug loading,
and are appropriate and safe for transdermal drug delivery. Ling and
his coworkers developed a rapidly dissolving MN patch composed of
starch and gelatin for the systematic transdermal delivery of insulin.
The MNs consisted of 225 (15 * 15) pyramidal needles with a height
of 600 μm, base width of 300 μm. They reported the starch/gelatin
MNs rapidly dissolved and released the encapsulated payload within
5 min after skin insertion. The results indicated that the insulin-loaded
MNs efficiently reduced blood glucose levels in diabetic rats with a rel-
ative pharmacological availability and relative bioavailability of 92.2 ±
3.5% and 92.3 ± 7.2%, respectively. Thus, dissolving starch/gelatin MNs
could be a promising approach for the transdermal delivery of various
biomolecules. The dissolution mechanism of rapidly dissolvable
starch/gelatin microneedles are presented in Fig. 3 [52].
3.4. Hollow microneedles
Hollow MNs use the “poke-and-flow” approach. They consist of a
space that is filled with the drug solution or dispersion. Holes are pres-
ent at the MN tips. Upon insertion, the drug is directly deposited in the
epidermal layer [9,20,32,41]. Norman et al. achieved targeted transder-
mal injection using poly(lactic acid) hollow MNs. Upon injection of the
MNs, 90% of the material was targeted within the skin with negligible
leakage onto the surrounding surface [53]. Mishra and co-workers fab-
ricated hollow MNs using SU-8 on a silicon substrate. The maximum
compressive force was found to be 0.27 N per needle, which allowed
successful penetration through the rat and mice skin without any break-
age. Microfluidic characterizations demonstrated hollow MN lumen de-
velopment with a flow rate of 0.93 μL per sec at 2 KPa pressure
difference at the inlet [54]. The unique characteristics of hollow MNs
such as delivering quantitative drugs at different pressure-driven flow
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

rates, delivering the drug directly into the epidermis, appropriate me-
chanical strength makes them potential candidates for transdermal
drug delivery.
3.5. Hydrogel forming microneedles
Hydrogel-forming MNs are a recent addition to the MN delivery sys-
tems. They are also known as swellable MNs. Super-swelling polymers
are used to fabricate the MNs. When swellable MNs are inserted into
the skin, the biological fluids are absorbed by the MNs because of the
polymeric cross-linking and hydrophilic groups. Thus, the high concen-
tration of the drug from the MNs is released into the circulation. The
major disadvantage of these MNs is poor mechanical strength. Attain-
ment of balance between the mechanical strength and swellability by
choosing appropriate polymeric material and process for fabrication of
MNs is required for optimal use of swellable MNs [7,32]. The advantages
and disadvantages of the different types of MNs have been listed in
Table 2 [7,9,20,32,41].
Considering the advantages of dissolving/ biodegradable MNs over
other types, the different materials used for their preparation and the
recent studies will be elaborated further. The current scenario of the
therapeutic applications of biodegradable MNs will be understood.
3.6. Biodegradable microneedles
Biodegradable MNs use different kinds of biodegradable polymers
including polylactic acid, polyglycolic acid, poly(lactide-co-glycolide)
(PLGA), or chitosan. The drug is encapsulated within the biodegradable
MNs, followed by insertion in the skin. After application, the
biodegradable polymer forms a matrix that degrades in the skin. Thus,
no residues from the MNs remain in the body [55,56]. Li et al. developed
solid polylactic acid MNs for pretreatment of the skin to enhance drug
permeation and absorption. Insulin was used as the model drug to
study in vivo activity in diabetic mice. Due to the increased permeability
of insulin upon MNs pretreatment, the blood glucose levels of the mice
were reduced to 29% of the administered level at 5 h [57]. Park et al. de-
veloped multilayered polymeric MNs composed of poly(lactic-co-
glycolic acid) (PLGA) and polyvinyl pyrrolidone exhibited controlled
drug release. Bovine serum albumin (BSA) was used as the model
drug, and spray deposition was used to form the multiple layers. In
vitro release studies demonstrated that BSA in the polyvinyl pyrrolidone
layer was released within 30 min, whereas 60% of BSA in PLGA was re-
leased within 1 h, followed by slow continuous release over 7 days. This
approach could be used to modulate the drug release profile using bio-
degradable polymers [58].
Biodegradable MNs can be easily fabricated using a variety of
methods such as microinjection, templating, or lithography at room
temperature. They are particularly suitable for delivery of drugs
[59–61], vaccines [23,62,63], DNA [44,64], and proteins. MNs can be
used to substitute conventional sustained-delivery systems. However,
they must remain in the skin to utilize the biodegradability of the poly-
mer effectively [48,55]. Recently, PLGA MNs were used to formulate in-
tradermal implants for etonogestrel (third-generation progesterone).
The pharmacokinetic studies in rats demonstrated that the plasma
etonogestrel level was detectable until 36 h. The AUC0→48h only
accounted for 37.8% of AUC0→∞, suggesting a sustained effect [65]. Bio-
degradable MNs can be used for the controlled intradermal release of la-
bile molecules such as enzymes [66]. Tsioris et al. developed silk MNs for

Table 2
Classification of microneedles [7,9,20,32,39].

Type of MNs Approach Characteristics Advantages Disadvantages

Enhances • Poor dose accuracy

• Mechanical • Requires healing after applica-
Creates microchannels in the skin. Upon subsequent application of strength tion
Solid MN Poke-and-patch
a drug patch, the drug enters skin with decreased resistance. • Physical sta- • Extends time of occlusion
bility • Reuse may cause infection
• Drug loading • Poor biocompatibility
Enhances
• Poor biocompatibility
• Stability of • Limited dose
Coated MN Coat-and-poke Enhance dissolution of biomolecules coated on the MN. drug • Peeling during insertion
• Mechanical • Formulation migration
strength
• Enhances sta-
bility of drug
• Low-cost
manufactur- • Limited dose
ing • Poor mechanical strength,
Dissolving MN Poke-and –release Fabricated with biodegradable polymers or sugars.
• Ease of biocompatibility, and physical
manufactur- stability
ing
• Controlled
drug release
• Sample
extraction • Clogging
Consists of a vacant space inside. Drug solution or dispersion can be
Hollow MN Poke-and-flow • Dose accuracy • Ingressing body fluids
filled in the hollow MNs.
• Constant flow • Requires prefilled syringe
rate
• Biodegradable
• Ease of
manufactur-
ing
• Poor mechanical strength and
Hydrogel-forming/swellable • Improves
– Absorb the biological fluids and swell. Excellent swellability. physical stability
MN drug loading
• Ingressing body fluids
• Controlled
drug release
• Easy
sterilization

607
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

the delivery of the horseradish peroxidase enzyme. The results revealed
that by controlling the secondary structure of silk protein, the drug re-
lease rate decreased by 5.6-fold [28].
The biodegradable MNs gained significance over the other MNs due
to advantages such as precise loading, intact removal of skin, and the at-
tachment of the drug reservoir. The natural biopolymers as a material
have been used in medical applications for centuries, and have gained
immense importance in drug delivery [67]. The following section dis-
cusses the various polysaccharides and polypeptides-based biomate-
rials used to fabricate biodegradable MNs. Additionally,
biocompatibility and biodegradation of these biomaterials and their ap-
plications in transdermal drug delivery will be discussed thoroughly.
4. Biodegradable microneedles: mechanism, natural polymers used
for their fabrication, and applications
Majority of the biodegradable MNs are manufactured using natural
biopolymers that degrade after skin insertion [67]. The degradation pro-
cess takes place over a prolonged time in a biological environment
under the influence of enzymes. The major advantages of this degrada-
tion include formation of nontoxic metabolites, and their elimination
from the body by natural metabolic pathways [68]. The degradation
process takes a longer time compared to dissolvable MNs. Hence, biode-
gradable MNs are gaining significance for the sustained delivery of ther-
apeutics. The release rate of incorporated therapeutic from these MNs is
managed by modulating the degradation rate of the chosen biodegrad-
able polymer [56].
The sources and properties of natural polymers such as polysaccha-
rides (e.g. cellulose, starch), and complex carbohydrate polymers that
make them suitable for the fabrication of biodegradable MNs are com-
piled in Table 3 [69–95].
4.1. Polysaccharides or carbohydrate polymers
The polysaccharides that are used to fabricate MNs include cellulose
and its derivatives, starch, and complex carbohydrate polymers such as
alginates, chitosan, chondroitin sulfate, xanthan gum, pullulan, and
hyaluronic acid.
4.1.1. Alginate
Alginate is a polyanionic polysaccharide comprising of β-D-
mannuronic acid and α-L-guluronic acid. Alginates are obtained from
algae and bacteria. The commercial source of alginates is brown algae
(comprising 40% of the dry matter). Durvillea, Sargassum, Macrocytis,
Eclonia, Ascophyllum, Laminaria, and Lessonia species are used for com-
mercial production. Bacteria belonging to Azotobacter, and Pseudomonas
species are also used for the production of alginates. These bacteria pro-
duce alginate-like polysaccharides as an extracellular material. How-
ever, this method is not economically feasible and is used only for
small-scale research studies [69–71]. Alginate is biodegradable and bio-
compatible. However, its use for the fabrication of MNs has been with-
held due to its poor mechanical strength. Despite this, initial efforts
suggested that alginate MNs can create microholes in the skin without
any deformation [72,96,97]. The “poke-and-release” strategy of alginate
MNs was found to be very helpful because it prevents MNs reuse, the
risk of biohazards, and cross-contaminations. Demir et al. developed so-
dium alginate MNs for the transdermal delivery of bovine serum albu-
min. They observed that the MNs patch increased the permeation of
bovine serum albumin across human abdominal skin by 15.4 fold com-
pared to the needle-less sodium alginate patch [97]. Cross-linked algi-
nate blended with maltose was used to prepare MNs for transdermal
delivery of insulin. The MNs exhibited enhanced mechanical properties
due to the addition of maltose [98].

Table 3
Carbohydrate polymers, their sources, and their properties that make them suitable for fabrication of microneedles.

Carbohydrate Sources Properties suitable for fabrication of MNs References

polymers

Alginate -Brown algae: Durvillea, Sargassum, Macrocytis, Eclonia, -Microhole creation in the skin without producing any deformation [69–72]
Ascophyllum, Laminaria, and Lessonia species Thickening agent
-Bacteria: Azotobacter, and Pseudomonas species
Cellulose and -Wood -Durability [73–75]
its derivatives -Algae: Chaetamorpha melagonicum, Valonia ventricosa -Transparency
-Bacteria: Acetobacter, Agrobacterium, Rhizobium, Sarcina -Elasticity
species -Viscosity
-Bacterial cellulose has higher water absorption and retention capacity, low
solubility, high mechanical strength, resistance to degradation, and nano-scaled
and uniform fiber network.
Chitosan Crustaceans such as crabs, shrimps, lobsters, and krill. -Dissolvable [76–79]
-Swellable
-Absorbs and retains large amounts of water
- Natural antibacterial property
-Wound healing
Chondroitin Porcine skin and rib cartilage, bovine trachea, and shark -Water soluble [80–82]
sulfate (CS) cartilage. -Possesses bioactivity
Hyaluronic acid -Animal sources: Rooster combs, bovine synovial fluid, human -Versatility [83–87]
(HA) umbilical cord, and vitreous humor of cattle. -Enhanced permeability
-Bacterial production: Streptococci sp., Agrobacterium sp. -Sustained release
Bacillus subtilis, and Escherichia coli. -Skin hydration
-In vitro: Enzymes from Streptococcus pyogenes and Pasteurella -Viscoelasticity
multocida. -Tissue re-epithelization
Sugars Sugar cane and beet are major sources. Other sources include -Non-toxicity [88–92]
honey, corn, fruits and vegetables. -Minimal cytotoxicity
-Affordability
-Water soluble
-Antimicrobial properties
Xanthan gum Bacteria: Xanthomonas campestris, Xanthomonas arboricola -Excellent solubility in water [93–95]
(XG) and Xanthomonas axonopodis. -Retards drug release
-Time independent release kinetics
-Viscosity enhancement

608
N. Dabholkar, S. Gorantla, T. Waghule et al.
4.1.2. Cellulose and its derivatives
Cellulose is the most abundant structural component of herbal cells
and tissues. The primary source of cellulose is wood. Commercially, cel-
lulose is obtained from wood or cotton. The other sources of cellulose
include algae and bacteria [73,74]. The cellulose obtained by biosynthe-
sis in living organisms consists of glucose monomers only that are
linked by β-(1,4) linkages [99].
The two important cellulose derivatives are cellulose ethers and cel-
lulose esters. Cellulose ethers possess commercially important proper-
ties of solubility, viscosity, surface activity, and stability [74].
Carboxymethyl cellulose (CMC) is one of the most important cellulose
ethers. It is highly soluble in water and produces a clear solution, mak-
ing it a pharmaceutically acceptable polymer [100]. Cellulose esters are
water-insoluble film-forming polymers. Examples of cellulose esters in-
clude cellulose acetate, cellulose acetate phthalate and cellulose nitrate
[74]. Park et al. mixed CMC with amylopectin to enhance the mechani-
cal and dissolution properties of MNs for the controlled release of drugs
[101]. Garcia et al. fabricated self-sterilizing CMC MNs loaded with silver
nanoparticles. These MNs can be used as vehicles for vaccine and drug
delivery [102]. The havoc caused by SARS-CoV-2 is being witnessed on
the global level. Kim et al. developed a MNs-assisted recombinant coro-
navirus vaccine using CMC (molecular weight: 90 kDa) by a three-stage
MNs manufacturing process. The results demonstrated that significantly
high SARS-CoV-2 IgG responses were detected as early as week 2 for all
subunit vaccines compared to control groups [24].
4.1.3. Chitosan
Chitosan is a marine polysaccharide obtained by chemical purifica-
tion of chitin. Chitin is extracted from crustaceans such as crabs,
shrimps, lobsters, and krill. Chitosan is obtained commercially by
deacetylation of chitin. Chitosan is also found in the cell walls of certain
fungi [76,78]. Chitosan is a linear biopolymer formed by D-glucosamine
and N-acetyl-D-glucosamine linked by β-(1,4) bond [76,77]. The molec-
ular weight of chitosan is between 300 and 1000 kDa (source depen-
dent). Properties such as molecular weight, degree of deacetylation,
and moisture content determine the stability and the mechanism of
degradation of a material. It was reported that the lower molecular
weight-based chitosan exhibits low mechanical strength. To overcome
the limitations of low mechanical strength of chitosan MNs, they were
mounted on to PLGA as a supporting array [103]. In another study,
thiolated chitosan was selected for MN fabrication, as the presence of
the thiol group exhibits high mechanical strength and high-water up-
take ability. This enhanced the overall swelling properties of the poly-
mer. The MNs prepared with 2% thiolated chitosan exhibited ease of
fabrication, and enhanced sharpness and tensile strength of the needle
compared to 1% and thiolated chitosan. High concentration (3%
thiolated chitosan) was difficult to pour and fill the micropores of the
template [104].
Chitosan is bioabsorbable. However, it is poorly soluble in aqueous
solutions at neutral pH. It is degraded slowly by lysozymes and
chitosanases. Chitosan behaves like a hydrogel because it absorbs and
retains large amounts of water, allowing it to swell without dissolving
completely [76]. Additionally, it possesses unique properties such as
natural antibacterial and wound healing properties [105]. Chen et al. de-
veloped chitosan MN patches using bovine serum albumin (BSA) as the
model drug. In vitro studies performed on porcine skin demonstrated
that BSA was released in a sustained manner for at least 8 days. Alexa
Fluor 488-labeled BSA MNs were applied on the rat skin in vivo. BSA dif-
fused gradually into the dermal layer and the fluorescence was ob-
served at a depth of 300 μm [106]. Chen et al. developed embeddable
MNs system using poly(L-lactide-co-D,L-lactide) as a supporting array
for sustained intradermal release of encapsulated antigens [103]. Sadeqi
et al. presented fabrication of solid and hollow chitosan MNs using the
cost-effective method of cross-over lines laser engraving [107].
Chandrasekharan et al. prepared trifluoroacetic acid-treated water-
soluble chitosan for MN-assisted delivery. The MNs patch showed
609
International Journal of Biological Macromolecules 170 (2021) 602–621
prolonged release of rhodamine B for >72 h. Therefore, these MNs can
be suitably used for the delivery of peptides [108]. Thiolated chitosan
can also be used for preparing MN patch [104]. Chi et al. developed a
temperature-responsive chitosan MNs patch for wound healing. This
smart delivery system is a novel approach that can be utilized in
wound healing [109].
4.1.4. Chondroitin sulfate
Chondroitin sulfate (CS) is a sulfated glycosaminoglycan consisting
of disaccharide units of d-glucuronic acid and N-acetyl-galactosamine
linked by β-(1,3) glycosidic bonds. There are two major CS, A and C, in
which the sulfate group is present at different positions. The sulfate
group is present in the 4th position in CS A and the 6th position in CS
C. The sulfate group provides hydrophilicity to CS [80,81]. CS is a
major component of cartilage and imparts elasticity to the cartilage.
CS can be predominantly obtained from porcine skin and rib cartilage,
bovine trachea, and shark cartilage [82]. The concentration and compo-
sition of CS depend on the source, terrestrial or marine, from which it is
obtained. CS exhibits antioxidant, anticoagulant, anti-inflammatory,
and anti-thrombus activities making it beneficial in joint-related pathol-
ogies [110]. CS is a water-soluble and biodegradable polysaccharide
[111]. Poirier et al. developed a dissolvable MN array composed of CS
and hydroxyethyl starch. It was loaded with hepatitis B surface antigen
with QS-21 saponin as an adjuvant. The antigenicity was retained at
37 °C and 45 °C. Only a 10% loss was observed at 50 °C after 6 months
[112]. Siqi Liu et al. fabricated recombinant Staphylococcal enterotoxin
B-loaded dissolvable MNs composed of CS (2%) and trehalose (0.8%).
The MNs easily penetrated the skin within 5 min with a skin penetration
depth of 260 μm. The MNs extended the antigen retention time in vivo
significantly [113].
4.1.5. Hyaluronic acid
Hyaluronic acid (HA) or hyaluronan is a linear, non-sulfated glycos-
aminoglycan. It consists of N-acetyl-D-glucosamine and D-glucuronic
acid linked by β-(1,4) glycosidic bonds [84]. The strength of MNs is an
essential characteristic for dermal penetration. The increase in the HA
molecular weight and concentration leads to reinforcement of the
three-dimensional polymer network [114].
HA is present in the extracellular matrix of connective tissue of most
vertebrates. It is synthesized by a class of proteins known as HA
synthases [115]. High concentrations of HA are found in the skin, umbil-
ical cord, synovial fluid of joints, and vitreous humor of the eye. 50% of
HA in the human body is present in the skin [83]. It has been found
that rooster combs contain 7.5 mg/g of HA, which is the highest re-
ported in an animal tissue [116]. HA is also present in the pathogenic
microbe Streptococci and non-pathogenic microbes including Enterococ-
cus faecalis, and Bacillus subtilis [83,117]. HA enhances the permeability
and helps in achieving sustained release. Zhu et al. have described the
possible mechanisms producing the aforementioned effects [84]. HA
possesses many unique advantages, allowing its use in TDDS. HA is a
suitable material for making MNs. This is because of its biocompatibility
and biodegradability. Also, because of its versatile nature, many differ-
ent formulations such as solutions, nanorods and nanostructured lipids
can be easily loaded [84,118–120].
HA based MNs were used to enhance lymphatic delivery via the
transdermal route. Lymphatic delivery can be used essentially for vacci-
nation and tumor chemotherapy [121]. Liu et al. used HA based MNs for
the transdermal delivery of insulin. It was found to be effective and safe
for use in the clinic [26]. A combination of various approaches can help
to optimize transdermal drug delivery. Hao et al. combined the ap-
proaches of near-infrared responsive PEGylated nanorods and HA MNs
for transdermal delivery of doxorubicin [120].
4.1.6. Sugars
Sugar, which commonly refers to sucrose, is a natural water-soluble
carbohydrate. Sugar can be obtained from various sources including
N. Dabholkar, S. Gorantla, T. Waghule et al.
honey, corn, fruits, and vegetables; sugar cane and beet being the com-
mercial sources [88]. Sugars are safe and affordable. Sugars also exhibit
antimicrobial properties against infection and wound causing patho-
gens [7,122]. The commonest sugar used to fabricate MNs is maltose.
Other alternatives include trehalose, galactose, sucrose, mannitol, and
xylitol. However, sugars present problem in the processing (require-
ment of thermal treatment), storage, and application on the skin
[89,90]. Yalcintas et al. conducted in vitro study to investigate the cyto-
toxicity of various carbohydrate-based materials such as glucose, treha-
lose, CMC, maltodextrin, and HA. The results obtained from different
qualitative and quantitative analyses suggested that trehalose, CMC,
maltodextrin, and HA do not pose cytotoxicity challenges and thus can
be used to make MNs. However, glucose should be used at low concen-
trations; as it was found to be toxic to the cells at concentrations above
50 mg/mL [91]. Milewski et al. developed MNs for the delivery of an in-
vestigational peptide using MicroCor®. The coating solution contained
the peptide and sugars (dextran and sorbitol) [123]. Sugars such as tre-
halose [63] and sucrose [124] have also been used to stabilize the for-
mulations for effective loading on the MNs.
4.1.7. Xanthan gum
Xanthan gum (XG) is a high molecular weight heteropolysaccharide
secreted by the bacteria Xanthomonas campestris. XG can also be ob-
tained from Xanthomonas arboricola and Xanthomonas axonopodis. The
bacteria produce XG at the cell wall surface by an enzymatic process
during its normal life cycle [93,94]. XG consists of β-(1,4)-D-
glucopyranose glucan backbone similar to cellulose with side chains of
(3,1)-α-linked D-mannopyranose-(2,1)-β-D-glucuronic acid-(4,1)-β-D-
mannopyranose on alternating residues [125]. XG possesses good bio-
compatibility. It is soluble in both hot and cold water [93]. XG causes re-
tardation of drug release and can offer time independent release
kinetics [94,125]. For coating MNs successfully, different coating
methods are used which require different viscosities and surface ten-
sion. Different excipients have been added to the coating solutions to
obtain these desirable properties. XG has been used to increase the vis-
cosity of the coating solution of MNs [95]. Increased viscosity leads to
improvement in coating thickness [126]. Kim et al. developed MNs
coated with influenza virus-like particle vaccine. XG (1% w/v) was
used as a viscosity enhancer in the coating solution. XG produced better
hemagglutinin activity than carboxymethyl cellulose. However, the
coating dose of XG was significantly lower compared to carboxymethyl
cellulose. Hence, carboxymethyl cellulose was used as the viscosity en-
hancer [127]. Choi et al. used XG (0.075% w/v) as the viscosity enhancer
with trehalose solution to prepare MNs coated with the whole
inactivated influenza vaccine. Due to the addition of XG, the hemagglu-
tinin activity increased significantly to 81% compared to trehalose alone
which was 50–60% [128].
4.1.8. Pullulan
Pullulan is a hydrophilic linear biopolymer obtained from yeast-like
fungus Aureobasidium pullulans and composed α-(1→6) maltotriose
residues. Due to the presence of more hydroxyl groups, the chemical
modification of pullulan made it easy. Thus, pullulan and pullulan deriv-
atives possess special characteristics like good film-forming ability, ad-
hesiveness, and excellent mechanical performance and plays a key
role in biomedical applications [129]. Vora et al. developed a dissolvable
MN of pullulan for transdermal drug delivery of small molecule (meth-
ylene blue and fluorescein sodium) and with biomolecules (fluorescein
isothiocyanate labeled bovine serum albumin and insulin). The ex vivo
porcine skin permeation studies demonstrated that 95–100% of MNs
penetrated, which is similar to results attained with solid MNs. The pre-
pared dissolvable pullulan MNs successfully delivered the small mole-
cule and biomolecules across neonatal porcine skin. Therefore, the
pullulan MNs patch could be efficient to penetrate the stratum corneum
[130].
610
International Journal of Biological Macromolecules 170 (2021) 602–621
4.1.9. Bletilla striata
Bletilla striata is a water-soluble polysaccharide extracted from tu-
bers of Bletilla striata. It is a glucomannan comprising of α-mannose,
β-mannose, and β-glucose. This polysaccharide is mostly used in the
fabrication of gels, adhesives, and scaffolds because of its non-toxicity,
less cost, good stability, biocompatibility and biodegradability proper-
ties [131]. Hu et al. developed rhodamine loaded dissolving MNs using
Bletilla striata polysaccharide. The texture analysis and histological ex-
amination demonstrated that the MNs had sufficient mechanical
strength to be inserted into the skin. After the removal of MNs, slight ir-
ritation appeared on the skin but recovered within 24 h. In vitro studies
indicated that the rhodamine B-loaded MNs could provide sustained re-
lease of the drug [132].
The summary of applications of MNs fabricated from polysaccha-
rides or complex carbohydrate polymers have been listed in Table 4.
4.2. Polypeptides or protein polymers
The polypeptide or protein biopolymers that are used to fabricate
MNs include, zein [30,158], collagen [159], gelatin [59,160,161], fish
scale biopolymer [60,162–164], and silk [28,165–168].
4.2.1. Zein
Zein is a water-insoluble prolamine protein found in maize. Zein is
generally recognized as a safe (GRAS) approved biodegradable polymer
and sustainable agropolymer. Recently, it has been used in the fabrica-
tion of fibers, textile, plastic, chewing gums, and an adhesive. Bhatnagar
et al. investigated the effect of physicochemical properties of chemo-
therapeutic agents (tamoxifen and gemcitabine) in loading behavior,
drug disposition within the skin, and permeation across the skin by
zein microneedles. The loading capacity for gemcitabine and tamoxifen
was found to be 1459 ± 74 μg and 607 ± 21 μg respectively. The skin
permeation study on porcine skin revealed that after 48 h application
of zein MNs, tamoxifen was retained on viable epidermis whereas the
gemcitabine microneedles showed the greatest percentage permeation
across excised skin via poke and patch approach [30].
4.2.2. Collagen
Collagen is a protein with amino acids such as arginine, glycine, pro-
line, and hydroxyproline. It possesses high mechanical strength. Due to
its non-immunogenic, non-toxic, biodegradable, and biocompatible na-
ture, it is mostly applied for tissue engineering [159]. Aditya et al. pre-
pared dissolvable collagen microneedles of tryptophan blue (model
drug) using a micro-molding technique. They investigated the effect of
process parameters like biopolymers w/v ratio, MN height, pressure
drop, and duration of solidification on the production of MN. They re-
vealed that the duration of the microneedle inside the vacuum oven in-
fluences the microneedle height. Similarly, increasing PVP
concentration increases solution viscosity and enhances drug loading,
but it makes it difficult to penetrate the mold cavities [159].
4.2.3. Gelatin
Gelatin is a mixture of proteins derived by acid hydrolysis (type A
gelatin) and alkaline hydrolysis (type B gelatin) of collagen. Gelatin
has a molecular weight ranging from 15 to 400 kDa. The low molecular
weight gelatin forms soft gels; high molecular weight gelatin provides
suitable strength to the MNs [169]. Gelatin is readily available, inexpen-
sive, and can be easily fabricated into different shapes. Recently gelatin
was studied in fields of bio-industry such as tissue engineering
(cell-interactive coatings), drug delivery, biological glues, and tissue
scaffolds. Gelatin is not utilized in fiber production because of its strong
hydrogen bonding and its potential to harden at low temperatures
[170]. Yu et al. prepared gelatin and hydroxyapatite MNs for the deliv-
ery of insulin. They reported that s.c. injection of insulin resulted in a
sharp increase of the plasma insulin concentration with the highest
value at 103.5 ± 9.5 μIU/mL in ~1 h, and it decreased rapidly to a
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

Table 4
Summary of applications of microneedles fabricated from polysaccharides and complex carbohydrate polymers.

Carbohydrate Model drug Application Results References

polymer

Alginate Bovine serum albumin (BSA) Peptide and protein delivery
Insulin Diabetes mellitus
Insulin Diabetes mellitus
Cellulose and Ascorbic acid (Vitamin C) Cosmetic products
its
derivative
Influenza vaccine Influenza vaccination
Vehicles for vaccine and drug
Silver nanoparticles
delivery
Finasteride (FNS) Androgenetic alopecia
Valproic acid (VA) Hair regrowth
Zika virus vaccines: Ad5.ZIKV-Efl and
Zika virus vaccination
ZIKV-rEfl
Local delivery of insoluble
Cyclosporin A (CyA)
drugs, Psoriasis treatment
Donepezil hydrochloride Alzheimer's disease
Chitosan Rhodamine B Transdermal delivery of
hydrophilic drugs and
proteins
Transdermal delivery of
Bovine serum albumin macromolecules such as
proteins
Ovalbumin Intradermal vaccination
Tetanus toxoid Immunization
Androgen deprivation
Leuprolide acetate
therapy for prostate cancer
Diphtheria toxoid Dermal immunization
Ovalbumin Low-dose immunization
– Solid or liquid drugs
Sustained delivery of
Tacrolimus
tacrolimus
MN patch increased the permeation of BSA across human abdominal [97]
skin by 15.4 fold compared to needle-less sodium alginate patch.
Alginate blended with HA was used to fabricate MNs. The relative
[133]
bioavailability of insulin from MN patches was found to be 92.9 ± 7%.
Alginate blended with maltose was used to fabricate MNs. The MNs
exhibited enhanced mechanical properties due to addition of maltose [98]
(highest failure force: 0.41 N/ needle approximately.).
Amylopectin was mixed with carboxymethyl cellulose (CMC) to [101]
enhance the mechanical and dissolution properties of MNs. Also, an
enhanced anti-oxidant activity of ascorbic acid was obtained.
1% sodium CMC was used to prepare MNs. MNs formulated with
calcium heptagluconate and arginine had no significant loss in activity [22]
for influenza vaccine during storage at RT for 6 months.
The CMC MNs were self-sterilizing. Zones of inhibition developed
within 24 h around nanosilver-loaded MNs but not around the plain [102]
CMC MNs.
CMC was used to prepare the base of MNs. Sustained release of the
implanted FNS powder for 3 days was obtained with only one [134]
application of FNS powder-carrying MNs.
12% CMC was used to fabricate MN backbone. The MNs delivered VA
with higher accuracy than topical application and stimulated wound [135]
re-epithelialization signals involved in hair follicle regrowth.
Pups born to mice immunized with Ad5.ZIKV-Efl were aprotected
against the infection without weight loss or neurological signs. Pups
[23]
born to dams immunized with MNA-ZIKV-rEfl were partially protected
(50%).
CyA MNs were fabricated using hydroxypropyl cellulose. The MNs
penetrated skin successfully with up to 50% CyA content. Upon [136]
administration of 10% CyA to rats, CyA MNs showed Tmax of 8 h, Cmax
of 15.9 ng/mL, and AUC of 686, compared to Tmax of 2 h, Cmax of
18.205 ng/mL, and AUC of 254 for oral administration of solubilized
CyA.
Carboxy methyl cellulose was used to fabricate the base of the MNs
and hydroxypropylmethyl cellulose was used to make the tips of MNs.
[137]
95% of the drug was delivered into the porcine skin after 5 min of
insertion.
The MN patch made of water soluble chitosan showed prolonged [108]
release of rhodamine B for >72 h. The MNs exhibited sufficient
mechanical strength.
MNs had sufficient mechanical strength for insertion at 250 μm and
200 μm in the porcine skin (in vitro) and rat respectively. In vitro
studies revealed that BSA was released in a sustained manner for at
[106]
least 8 days. In vivo studies demonstrated BSA diffusion gradually into
the dermal layer and the fluorescence was observed at a depth of
300 μm.
Upon embedding the ovalbumin-loaded MNs in rat skin in vivo, the
MNs degraded gradually and prolonged ovalbumin exposure for upto
[103]
14 days. Compared to i.m. vaccination, the MNs demonstrated higher
ovalbumin-specific antibody response that lasted for at least 6 weeks.
In vitro studies demonstrated better skin penetration following
immunization by MNs. In vivo studies demonstrated comparable
induction of IgG and IgG1 titer and higher IgG2a titer by MNs [138]
compared to commercial tetanus toxoid. Also, MNs generated higher
Th1 cytokines.
The MNs can be loaded with about 73.27±2.75 μg of the drug. The
penetrated wound in Switzerland albino mice healed completely
within 7 days. After applying the MNs, serum luteinizing hormone
level increased and declined within the first 7 days. However, serum [139]
testosterone levels increased, reached a peak on the 14th day and
declined on the 21st day. The castration level can be maintained for
2 weeks.
The layer-by-layer coated pH-sensitive MNs were more efficient than
s.c. injection. The EC50 dose of the toxoid was 3 fold lower upon the [140]
use of MNs compared to s.c. injection.
Rats immunized with low-dose ovalbumin (200 μg)-loaded MNs had
high levels of antibody levels for 18 weeks persistently, demonstrating [141]
2.5 fold sparing in the i.m. injection dose (500 μg) of ovalbumin.
Chitosan lactate MNs can load large quantities of drug and
demonstrate good puncture performance. There is no need of chemical [142]
modification of the drugs before loading.
The MN patch demonstrated 84% of skin penetration without any
breakage. In vitro and ex vivo permeation studies demonstrated [104]
sustained release (85%) of tacrolimus from MNs compared to controls.

(continued on next page)

611
N. Dabholkar, S. Gorantla, T. Waghule et al.
Table 4 (continued)
Carbohydrate Model drug Application
polymer
Vascular endothelial growth factor Wound healing
Cancer chemo-photothermal
Doxorubicin and AuMSS nanorods
therapy
Chondroitin Staphylococcal enterotoxin B (SEB) Staphylococcal Enterotoxin B
sulfate (CS) toxic shock syndrome and
food poisoning
Hepatitis B surface antigen Hepatitis B vaccination
Leuprolide acetate (LA) Prostate cancer
H1N1 and H5N1 influenza
Split virion and whole virus particle
virus vaccine
Neurotoxin Rheumatoid arthritis
Ovalbumin Percutaneous vaccination
Insulin Diabetes mellitus
Recombinant human growth hormone
Peptide/ protein delivery
(rhGH) and desmopressin
Transdermal delivery of
Sinomenine hydrochloride (SH)
sinomenine hydrochloride
Blood glucose monitoring
– [149]
device
Hyaluronic Magnesium ascorbyl phosphate (MAP) Transdermal delivery of MAP
acid (HA)
Hyaluronic acid-antigenic peptide Prophylactic cancer
conjugate immunotherapy
Insulin Diabetes mellitus
Systemic delivery of
Artemether
poorly-water soluble drugs
Human epidermoid cancer
Doxorubicin
therapy
Polydatin/hydroxypropyl-β-cyclodextrin
Acute gout arthritis
inclusion complexes
Photodynamic therapy of
5-Aminolevulinic Acid
superficial tumors
Methotrexate Psoriasis
-Chemotherapy of tumor
Doxorubicin
-Immune-therapy for tumor
Topical steroids Prurigo nodularis
Sugars Human IgG Monoclonal antibody
delivery
Sinomenine hydrochloride Transdermal delivery of
612
International Journal of Biological Macromolecules 170 (2021) 602–621
Results References
Immunostaining demonstrated excellent anti-inflammatory effect and
[109]
remarkable collagen deposition compared to control.
The layer-by-layer MN patches of poly (vinyl alcohol) and chitosan
demonstrated a good photothermal capacity causing a 12 °C increase
in temperature under near-IR (808 nm, 1.7 W/cm2 for 5 min). In [143]
combination with the pH sensitivity of chitosan, it can be used to
control release of doxorubicin.
The dissolvable MNs were composed of CS (2%) and trehalose (0.8%). [113]
The MNs easily penetrated the skin within 5 min with a skin
penetration depth of 260 μm. The MNs extended the antigen retention
time in vivo significantly.
The antigenicity was retained at 37 °C and 45 °C. Only a 10% loss was
[112]
observed at 50 °C after 6 months.
Sodium CS was used to prepare MNs. In in vitro conditions, LA was
stable in the MN patch and 102.2 ± 1.9–95.3 ± 1.9% recoveries were
[31]
obtained for 3 months. In rats, the relative bioavailability of LA from
the MNs against s.c. solution was 105.6 ± 13.5%.
CS was used in the coating solution of MN. The MN had the potential to
enclose the dose of 15 μg of hemagglutinin, which is the conventional [144]
dose of vaccine (data not shown).
The ex vivo evaluation demonstrated that the penetration depth of
MNs in rats was higher than 70 μm. The cumulative penetration of
neurotoxin in MNs could reach 95.8% in 4 h. In contrast, the [145]
neurotoxin solution could hardly penetrate skin. The MNs were stable
for 3 months.
Insertion of the MNs delivered ovalbumin into the skin within 3 min
[146]
effectively.
The bioavailabilities of insulin from MNs were 72.1 ± 11.6% (for 2
patches) and 72.4 ± 8.3% (for 4 patches). Insulin was stable in the MNs [25]
for 1 month at 4 °C. The % recovery was 99.2 ± 13.9%.
The bioavailability of rhGH and desmopressin from MNs was found to
be 72.8 ± 4.2–89.9 ± 10.0% and 90.0 ± 15.4–93.1 ± 10.3%, respectively. [147]
The MNs were stable for one month at −80 °C and 4 °C.
The AUC, Tmax, and Cmax of SH-loaded MNs were found to be 33.07 ±
1.62 μg·h·mL−1, 9.74 ± 0.95 h, and 2.19 ± 0.72 μg·mL−1, [148]
respectively.
The adhesion time increased. The coefficient of determination R2
increased to 0.750.
Enhanced permeation of MAP using MN (96.8 ± 3.9 μg/cm2) [86]
compared to passive delivery of MAP (44.9 ± 16.3 μg/cm2).
The conjugates loaded into the MNs were localized near the
administration site and were retained for >24 h. A single vaccination
[150]
with MN patch resulted in a statistically significant tumor growth
inhibition in B16 melanoma mice model.
PK parameters; Cmax, Tmax and AUC of MNs was similar to that of
subcutaneous injection. After storage for one month at −40, 4, 20, and [26]
40 °C, >90% insulin remained in the MNs.
Maximum amount of artemether delivered into skin was 72.67±2.69%
[151]
of the initial dose that was loaded on MNs.
Remarkable anti-tumor efficacy; no recurrence was observed. Good
heating transfer efficacy was observed. The temperature of tumors [120]
treated with MNs rise to 60 °C within 1 min.
40% HA was used to fabricate MNs. The complexes increased the
aqueous solubility of polydatin to 124.47 mg/mL. The combination of
[152]
inclusion complexes and MN approach enhanced transdermal delivery
of polydatin.
Long term stability of MNs. In vivo studies demonstrated that in 3 out
of 5 mice, tumors continued to shrink and completely disappeared at [153]
day 14.
Alleviation of psoriasis-like skin inflammation in mice. Methotrexate
[154]
MNs were more efficacious than same dose administered orally.
MN enhanced bioavailability of doxorubicin. The Cmax value of
doxorubicin-loaded transferosome in MN was 153.3 ng/mL which was
3 folds higher than only transferosome. The absolute bioavailability of [121]
DOXQ loaded transferosome in MN was 79.9%; 3 times higher than
epidermal diffusion of only transferosome.
The pruritus visual analogue scale was significantly lower for MN
compared to topical application. Topical steroid application followed
[85]
by application of MN patch could be a useful strategy for treating
refractory skin diseases.
Maltose MNs: Methylene blue was taken up by microchannels [155]
indicating disruption of the stratum corneum and reaching the dermis.
Immunohistochemical studies demonstrated that IgG follows
microchannels for transport across the skin.
Maltose MNs: In vivo transdermal penetration studies of hydrogel and [61]
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

Table 4 (continued)

Carbohydrate Model drug Application Results References

polymer

sinomenine hydrochloride MN arrays demonstrated the following pharmacokinetic parameters;

for rheumatoid arthritis AUC0−t and AUC0−inf were 5.38 ± 0.53 μg (mL h)−1 and 35.35 ±
1.27 μg (mL h)−1, 5.75 ± 0.49 μg (mL h)−1 and 45.36 ± 2.36 μg
(mL h)−1, respectively. The MN array had slight irritation to the rat
skin.
25% w/v trehalose: The studies conducted in rhesus macaques
indicated that MNs are similarly immunogenic as i.m. vaccination
Adjuvant-free hepatitis B vaccine Hepatitis B vaccination [62]
without adjuvant. However, MNs are less immunogenic than i.m.
vaccination with alum adjuvant.
Trehalose MNs: % drug recovery of pentagastrin and sincalide
Pentagastrin and sincalide analogues Peptide delivery analogues from the MNs was 89.4 ± 10.61% and 85.7 ± 7.6% compared [156]
to 56.4 ± 9.37% and 90.2 ± 7.7%, respectively in the control.
Trehalose MNs: MNs retained most of the activity after storage for up
Human growth hormone Biopharmaceuticals delivery to 15 months at RT. After insertion into rat skin, pharmacokinetics of [27]
human growth hormone were similar to conventional s.c. injection.
Trehalose MNs: The MN patch-induced neutralizing antibody
Trivalent Sabin inactivated poliovirus
Polio vaccination responses were comparable to higher vaccine doses delivered i.m. for [157]
vaccine
type 1 and type 3 poliovirus serotype.
Xanthan gum Influenza virus-like particle vaccine Influenza vaccine XG (1% w/v) was used as a viscosity enhancer in the coating solution. [127]
(XG) XG produced better hemagglutinin activity than CMC. However,
coating dose of XG was significantly lower compared to CMC.
XG (0.075% w/v) was used as the viscosity enhancer with trehalose
solution to prepare MNs coated with whole inactivated influenza
Whole inactivated influenza vaccine Influenza vaccine vaccine. Due to the addition of XG, the hemagglutinin activity [128]
increased significantly to (81%) compared to trehalose alone
(50–60%).
Pullulan Insulin Diabetes mellitus The secondary structure of insulin was maintained upon storage at [129]
4 °C, 20 °C, and 40 °C. The MNs delivered 87% of insulin, 120 min after
insertion into human abdominal skin in in vitro.

lower level within 4 h. In contrast, prepared MN patches application ex-
hibited the highest plasma insulin level (190.7 ± 9.6 μIU/mL) for 2 h for
high loading insulin at 20 IU. Thus, the plasma insulin level could be
maintained for a longer time using MNs compared to s.c. injection [160].
4.2.4. Fish scale polymer
The fish scales naturally consist of protein polymers such as collagen
fibrils, which are arranged in orthogonal plywood structure in a hy-
droxyapatite matrix. The fish scale biopolymer is extracted as collagen
peptides from the skin surface of fishes by enzymatic, acid, and thermal
hydrolysis. The biopolymers are water-soluble and possess the excellent
film-forming ability. Hence these can be used in the micromold fabrica-
tion [162]. Olatunji et al. developed a methylene blue loaded fish scale
polymer extracted from the tilapia (Oreochromiss sp.). They reported
that fish scale MNs successfully penetrated the porcine skin and dis-
solved gradually at 0, 60, 120, and 180 s after insertion. The mechanical
analysis study demonstrated that the MNs with tip radius between 10
and 100 μm could withstand 0.12 N of force per MN without any defor-
mation [162].
4.2.5. Silk fibroin
Silk fibroin is well known purified biocompatible polymer obtained
from the proteins produced by Bombyx mori [171]. Silk fibers are usually
polyamino acid-based fibrous proteins. Silk proteins have acquired im-
portance in the medical field because of their innate properties like good
mechanical strength, resilience in the biological environment, biocom-
patibility, and biodegradability. Silk has been applied in various biomed-
ical fields like fibers, scaffolds, and hydrogels [172]. You et al. prepared
methylene blue loaded rapidly dissolving fibroin MNs). The results re-
vealed that silk fibroin MNs dissolved within 1 min under the skin to re-
lease the drug. The dissolved fibroins were sharp and strong enough to
penetrate the porcine skin and generated non-inflammatory amino acid
degradation products that are used in cell metabolic functions [165].
The summary of applications of MNs fabricated from poly-
peptides or protein polymers have been listed in Table 5
[28,59–60,158,161,163,166–168].
4.3. Polysaccharide and polypeptide composites
The polysaccharides and polypeptides with lower molecular weight
may not exhibit the proper mechanical strength for dermal penetration.
It has been reported that higher molecular weight provides better
strength, and can help attain the required MN geometry [173]. In a
study, CMC arrays on thermal treatment led to decreased water content,
forming hydrogel in the mold. Moreover, continuous heating of the hy-
drogel led to shrinkage. Upon increase in the concentration of CMC, hy-
drogel formation was faster with higher shrinkage. The alginate arrays
disclosed further shrinkage compared to HA and CMC. In the reported
study, MNs prepared using CMC alone were not adequate for use. The
needles formed were fragile with significant deformation on application
of minimum weight (5 N). The addition of amylopectin increased me-
chanical strength of the MNs. The higher concentration of amylopectin
resulted in enhanced mechanical strength. MNs mixed with amylopec-
tin hold >90% of the original height [101,174]. Thus, polysaccharide and
polypeptide composites were considered as an attractive solution to in-
crease the molecular weight of the polymers, and obtain the desired
mechanical strength and MN geometry.
In another study, Zhang and co-workers fabricated MNs from algi-
nate and maltose for the transdermal delivery of insulin. The MNs
were fabricated by the template method. To enhance the mechanical
strength of the MNs, 15% w/w maltose monohydrate was added. The
MNs exhibited strong mechanical properties with the highest failure
force of 0.41 N per MN. The MNs were then applied for the transdermal
delivery of insulin on diabetic Sprague-Dawley rats. Compared to the
subcutaneous injection, the relative bioavailability of the composite
MNs was found to be 93.7 ± 4.7% [175].
Cheng et al. fabricated HA composite MNs loaded with curcumin mi-
celles. Different excipients were used to increase the plasticity of HA and
facilitate the formation of MNs, such as polyvinyl pyrrolidone, gelatin,
and sodium carboxymethyl starch. It was found that the HA composite
MNs containing gelatin had low hardness and mechanical strength.
The HA composite MNs containing polyvinyl pyrrolidone were brittle
and tended to fracture. Ultimately, sodium carboxymethyl starch was
selected as the composite for HA MNs. The concentration of HA solution

613
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

Table 5
Summary of applications of microneedles fabricated from polypeptide or protein polymers.

Polypeptide Model drug Application Results References

polymer

Immunization studies demonstrated significantly (p < 0.001) greater antibody titers (IgG1, IgG2a, and total
Zein Ovalbumin Vaccination [158]
IgG) after the application of MNs compared with the control.
MNs were composed of gelatin and calcium sulfate. Excellent mechanical strength that can reach upto 0.4 N
Diabetes
Insulin for each MN. Compared to s.c. injection, hypoglycemic effect was observed for longer time in vivo because [161]
mellitus
of the MNs.
Gelatin
Gelatin methacryloyl was used to fabricate MNs. MNs without crosslinking released over 80% of
Doxorubicin Cancer doxorubicin in 30 min. Only a quarter of doxorubicin was released in MNs that were crosslinked for 60 s. [59]
Thus, the rate of drug release can be regulated by adjusting the degree of crosslinking.
Low temperature press method was used to fabricate MNs. The MNs could withstand up to 0.136 N of force
Methylene blue -
per MN. The cross-linked MNs swelled to 340% of their original mass in phosphate buffer solution. Within [163]
Ferrous gluconate -
Fish scale 24 h, 34.5% of the loaded drug was released, while the non cross-linked MNs dissolved within minutes.
Franz diffusion cell study in porcine skin demonstrated that the drug permeation rate increased from 2.5 to
Lidocaine – [60]
7.5% w/w after 36 h. Pseudo steady-state profile is observed from 5.0 to 10.0% w/w lidocaine-loaded MN.
Horseradish
The drug release rate decreased by 5.6-fold by controlling the silk protein secondary structure. Antibiotic
peroxidase Enzyme delivery [28]
loaded silk demonstrated a 10-fold reduction of bacterial density after MN application.
enzyme
Sustained
Insulin delivery of In vitro release studies demonstrated that the release time of drug from MNs was maintained up to 60 h. [166]
Silk fibroin
insulin
In vitro studies indicated that 2-ethoxyethanol modified silk MNs easily pierced porcine skin with a depth of
– – [167]
~200 μm and transformed into semisolid hydrogels with 50–700 nm porous network.
Methanol treatment enhanced the mechanical strength of the MNs up to about 200% depending on the
– – [168]
treatment duration. Methanol exposure time can effectively control drug release rates from the MNs.

was selected as 200 mg/mL and the proportion of HA to sodium
carboxymethyl starch was selected as 2:1. The MN matrix thus prepared
had satisfactory viscosity and fluidity and excellent mechanical proper-
ties. The MNs had sharp tips with a height of 600 μm, with regular ap-
pearance, without bubbles or broken MNs [176].
Chiu et al. developed composite MNs of sodium HA tip and chitosan
base to release biphasic antigen. Upon insertion of the composite MNs
in the skin, the sodium HA tip dissolved within the skin and rapidly re-
leased the antigen acting as a primer. The chitosan base remained in the
dermis, prolonging the antigen's release for 4 weeks, acting as a booster
dose. It was observed that single immunization with the composite MNs
containing ovalbumin, both the T helper type 1 and 2 were stimulated in
rats. When compared to the traditional subcutaneous vaccination, the
MNs produced higher and more durable antibody response. Thus, the
unique rapid and sustained release properties of the composite MNs
allow its use as a primer and booster vaccine formulation [177]. Ling
et al. fabricated dissolving composite MNs of starch and gelatin for the
transdermal delivery of insulin. Upon insertion into the skin, the MNs
dissolved within 5 min and quickly released insulin. Histological exam-
ination demonstrated that the MNs could be inserted into porcine skin
to a depth of 200 μm approximately in vitro, and 200–250 μm depth
into the rat skin in vivo. Compared to subcutaneous injection, the com-
posite MNs demonstrated relative pharmacological availability and bio-
availability of 92%. Even after storing the MNs at 25 °C or 37 °C for
1 month, >90% of the loaded insulin remained in the MNs. Thus, the
starch and gelatin composite MNs allowed rapid release of insulin [178].
5. Biocompatibility and biodegradation of the polysaccharide and
polypeptide MNs
The biocompatibility of MNs means that the MNs do not elicit any
undesirable local or systemic responses in the body. The accumulation
of the polymers at the tissue site and slow degradation is detrimental.
Thus, if the biopolymers used in MNs fabrication are degrading in the
body, their byproduct should be non-toxic. As the MNs penetrate bio-
logical barriers and come in contact with viable tissue, the appraisal of
biocompatibility of these MNs is essential.
Majority of the polysaccharides or carbohydrate polymers are ob-
tained from natural sources. Moreover, the structural resemblance of
glycosaminoglycans (GAGs) with skin components has led to the
intuition of good biodegradability and biocompatibility [179].
Hyaluronic acid and chondroitin sulphate are GAGs, which are the
major components of the extracellular matrix in the skin and the carti-
lage of joints. Theydegrade in the body by lysosomal enzymes and free
radicals in the tissue matrix. Further, hyaluronic acid-based MN array
MicroHyala® are commercially available for cosmetic use
[173,180,181].
The cellulose ethers and cellulose esters are non-toxic, biocompati-
ble, biodegradable, and are extensively used in tissue engineering, diag-
nostics, drug delivery, and wound healing, enabling application of these
biopolymers as MNs. Similarly, alginate is a novel hydrogel biopolymer
that has gained application in tissue engineering and drug delivery
[182].Chitosan is biocompatible, but its clearance is based on molecular
weight. Chitosan of low and medium molecular weight can be degraded
into monomers by N-acetyl-beta-D-glucosaminidase and cleared by the
kidney. In contrast, chitosan of high molecular weight can be degraded
into suitable fragments by proteases for renal clearance [183]. Dextran
is a high molecular weight polymer degraded by amylases and elimi-
nated via kidney [184]. Starch-based amylopectin is biocompatible,
but does not degrade quickly in the body. As the amylose content of
starch increases, the elongation strength also increases. However, starch
has not been explored much for the fabrication of MNs [185]. Xanthan
gum is biocompatible as it is used in the food, cosmetic, and pharmaceu-
tical fields. Marketed by Cargill under the name Santiaxane™, this poly-
mer has been used as a controlled release agent in solid dosage forms,
and as a thickening agent in semi-solid, liquid and topical formulations
[186,187]. Pullulan is degraded by α-amylase enzyme, which is mainly
secreted by salivary glands and pancreas. Pullulan was declared as GRAS
and gained significance in the cosmetic market (skincare) due to its
non-toxic, biocompatible, and eco-friendly nature. As it resembles the
natural extracellular matrix systems, it has been used in the preparation
of scaffolds, films micro/nanoparticles, and fabrication of new matrix
systems, and now as MNs [188].
Protein-based polymers such as collagen, gelatin, and zein are cur-
rently used in pharmaceutical industry as drug delivery carriers. Gelatin
is a favorable material for MNs fabrication due to its moderate to low an-
tigenicity and non-carcinogenic nature. Also, gelatin has been widely
used in the preparation of soft gelatin capsules. Collagen is biodegrad-
able and biocompatible. It swells upon contact with water, but can
only be digested by specific collagenases and pepsin-cleaving enzymes

614
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

[189]. Similar behavior and degradation were observed with gelatin, as
it is an irreversible hydrolyzed form of collagen. A report demonstrated
that collagen exhibits poor stability and nonspecific immune reactions
in vivo. But in the case of gelatin, the absence of an aromatic ring
makes it non-immunogenic. Thus, gelatin has been used to fabricate
MNs in combination with other biopolymers [190,191]. In recent days,
zein has gained significance in bone tissue engineering and MNs fabrica-
tion. Zein has been recognized as GRAS, and possesses low immunoge-
nicity [192,193]. Silk is an attractive scaffolding material suitable for
tissue regeneration due to its biocompatibility. Seri® Surgical Scaffold
(Allergan, Inc. MA) is a silk-based biomaterial approved by FDA. The
preclinical biological testing of the scaffold confirmed its overall bio-
compatibility, non-toxicity, pyrogenicity, and allergenicity [194]. All
these instances confirm the biocompatibility and biodegradation of
these natural polymers, and their use in the fabrication of MNs.
6. Clinical applications
Currently, little is known of the long-term effects caused due to re-
peated insertion of MNs in the skin. It is important to know the dimen-
sions of micropores created by MNs and the time required for skin
barrier recovery to successfully commercialize MNs [195]. For this, it is
necessary to conduct studies in human subjects [196]. Only a few clini-
cal studies have been conducted using MNs. The clinical studies that
have employed MNs have been compiled in Table 6 describing
their phase and current status of the trial [197]. Other clinical
studies involving biodegradable MNs have been listed in Table 7
[85,119,198–199,200–201].
7. Patent status
The use of natural polymers has gained momentum since past few
years. The patents for biodegradable MNs involving the use of natural
polymers are few. Among these, the majority of patents have been ap-
plied for hyaluronic acid.
Russian inventors developed a microstructure for transdermal ad-
ministration and its method of production (RU2701361C1). The appli-
cation was granted in September 2019. They claimed a microstructure
containing a biocompatible polymer, selected from HA, CMC, alginic
acid, pectin, carrageenan, chondroitin (sulfate), chitosan, etc. The adhe-
sive material was selected from ethyl cellulose, hydroxymethyl cellu-
lose, etc. The aspect ratio of the microstructure was 1:5 to 1:2. They
used a microform to fabricate the microstructures. The microstructures
increased the rate of absorption of therapeutic agents into the skin. Ac-
cording to the invention, the D-type microstructure maximally in-
creased the mechanical strength of the microstructure due to the use
of triple structure [202]. A Japanese inventor developed a hyaluronic
acid microstructure with excellent solubility characteristics
(JP2019523680A). The inventor claimed that the microstructure had
an excellent dissolution rate in the skin which was regulated by
adjusting the ratio of crosslinked HA and therapeutic agent. The content
of the crosslinked HA and non-crosslinked HA was 5 to 100% (w/w) and
0 to 95% (w/w), respectively. When the crosslinked HA was 15–30% of
the total weight of HA, the dissolution rate was 50–60%, 65–75%, and
70–85% after 10 min, 30 min, and 60 min, respectively after MN applica-
tion. The delivery system can be used as a transdermal transmitter
[203]. A patent was filed for a microneedle applicator by a Russian in-
ventor (RU2016144039A). The application was granted in April 2018.
The invention claimed a microneedle applicator consisting of a
microneedle template, containing biodegradable microneedles loaded
drug. The microneedle applicator can be used for loading therapeutic
drugs, vaccines, or cosmetic preparations [204]. Zhongli Ding and co-
workers developed a microarray for delivery of therapeutic agent and
method of preparation of the same (EP2934660B1). The application
was granted in July 2019. The microstructure comprised of a backing,
and microstructure array. The microstructures consisted of a distal
layer composed of a biodegradable polymer and a proximal layer. The
therapeutic agent was loaded on the distal layer. The biodegradable
polymer was selected from about 50–100% of a hydrophobic polymer
with a glass transition temperature of about 20–25 °C [205].

Table 6
Microneedles currently in clinical trials.

Clinical Trials. Type of MN Drug molecule Primary outcome/description of study Phase Status
gov Identifier

NCT02955576 Microneedle HA patch Calcipotriol - The improvement of psoriasis condition in the time frame of two weeks NA Recruiting
betamethasone
dipropionate
NCT02682056 Biocompatible polymers – A comparative study for blood glucose level testing in patients with diabetes NA Completed
using microneedle patches, intravenous (IV) catheter draw, and lancet.
NCT03207763 Solid conical shape Placebo Vaccine delivery targeting the antigen-presenting cells present in the skin NA Completed
Microneedles made of
water-soluble excipients
NCT03203174 Microneedle; Type not Botulinum Toxin The primary purpose is to treat palmar hyperhidrosis Phase 1 Completed
available Type A
NCT00837512 Hollow Insulin To treat Type 1 Diabetes III Completed
NCT02594644 Stainless steel MNs (MR200, Aminolevulinic To treat Keratosis, Actinic. The results revealed that after the 20-min NA Completed
Clinical Resolutions Acid incubation arm, average actinic keratosis clearance was 76% vs 58% on the
Laboratory, Inc.) sham side (P< 0.01), which is not significant
NCT02995057 Gold- or silver-coated, or – The purpose of this study is the safety demonstration of microneedle, i.e., NA Completed
uncoated nickel microneedles biocompatibility and inertness. Participants have proven nickel allergy.
NCT00539084 MicronJet from nanopass Local anesthesia The primary purpose is to achieve painless delivery of drugs directly into the NA Completed
i.e., Lidocaine skin's superficial layers using the MicronJet microneedle device compared to
insertions of intravenous catheters.
NCT03607903 MicronJet600 microneedle Adalimumab A Randomized, Double-Blind, Placebo-controlled, Double-dummy Study to Adalimumab Active, not
from NanoPass Assess Microneedle Delivery in Comparison to Subcutaneous Injection of ID - II recruiting
Adalimumab in Healthy Volunteers.
Adalimumab
SC - III
NCT03472235 Dermapen – To treat Melasma. The primary outcome measures exhibited that no NA Not yet
improved efficacy recruiting
NCT02596750 Microneedle Roller (MR2 4% lidocaine To evaluate the role of microneedle pretreatment in the speed at which NA Completed
roller, Clinical Resolution anesthesia develops after applying topical 4% lidocaine.
Laboratories, Inc.)

615
N. Dabholkar, S. Gorantla, T. Waghule et al. International Journal of Biological Macromolecules 170 (2021) 602–621

Table 7
Clinical studies and outcomes of biodegradable MNs fabricated from natural polymers.

Year Natural Application About the study MN treatment (number of Other/control Major outcome of the study References
polymer human subjects) treatment (s) group
used to (number of human
fabricate subjects)
MNs

2014 Hyaluronic Skin Double-blind, Twice daily for 12 weeks (24) None (0) Both the retinyl retinoate- and ascorbic [198]
acid wrinkles randomized controlled acid-loaded dissolving MN patches
trial demonstrated statistically significant
differences in all Visiometer R-values (P <
0.05). In particular, highly significant
differences were shown at R1 (skin
roughness) and R5 (arithmetic average
roughness) (P < 0.001).
2017 Hyaluronic Psoriatic Ethics Committee of the Once daily at night for 1 week None (0) The mPASI (mean Psoriasis Area Severity [199]
acid plaques Catholic Medical Center (10) Index) was 5.4 (± 1.2) before treatment
Office of Human and 1.8 (± 1.1) after treatment (P < 0.001).
Research Protection The median decrease in mPASI was 60%
(VC17RESI0010) (range: 25–100%) after 1 week application
of the MN patch.
2017 Hyaluronic Crow's feet IRB of P&K Skin Research 20 min twice a week for HA essence group, Before the treatment, the global visual [119]
acid wrinkles Center P1512–55 8 weeks (31) twice a week for wrinkle score was 2.3±0.56 on the HA MN
8 weeks (31) group and 2.35±0.49 on the HA essence
group. At 8 weeks, the score of the HA MN
group was 2.32±0.54 and of the HA
essence group was 2.35±0.55.
2018 Hyaluronic Prurigo IRB No. 4–2015-1086 Mometasone furoate 0.1% cream Mometasone Investigator global assessment (IGA) score [85]
acid nodularis was applied two times per day furoate 0.1% cream demonstrated that weekly application of
for four weeks; followed by the was applied two MN after topical steroids was more
MN patch once per week for times per day for effective than topical steroids alone (2.33
12 h (24) four weeks (24) ± 0.23 for MN; 1.33 ± 0.24 for control)
(paired t-test, p<0.05)
2019 Hyaluronic Skin aging Approved by Twice a week onto the face in None (0) The averaged wrinkle depths before and [200]
acid International Medical the left and right corner of the after application of the product were 363 ±
and Dental Ethics eye and on a defined area on the 146 μm and 287 ± 166 μm, respectively.
Commission GmbH right volar forearm for 12 weeks After the treatment, an average increase of
(Freiburg, Germany) (20) 26% overall improvement of the wrinkle
depths was observed.
2020 Hyaluronic Crow's feet Randomized Once every 3 days, in the Once every 3 days, After 8 weeks of use, the maximum depth [201]
acid wrinkles double-blind clinical evening, for 8 weeks (23) in the evening, for of biggest wrinkles improvement in the
trial 8 weeks (23) Ad-HMN (high molecular weight HA
dissolving MN patch) group and (low
molecular weight HA dissolving MN patch)
group was 23.34% (P < 0.001) and 13.32%
(P < 0.01), respectively.

IRB: Institutional Review Board.

David L. Kaplan, and teammates filed a patent for silk fibroin based
microneedles and methods of fabricating the same (CA2815285C). The
application was granted in December 2019. They claimed that the
microneedle device, extending from the base to the tip, composed of
silk fibroin was highly biocompatible and easy to fabricate. The silk fi-
broin remains stable at room temperature and is implantable. They
stated that the rate of delivery of active agent can be controlled by reg-
ulating the structure of silk fibroin. These microneedles can be used to
deliver active agents into or across biological barriers such as cell mem-
brane, tissue, and skin. The method for fabrication of the microneedles
used micromolds [206].
8. Future prospects
The fabrication of biodegradable MNs using biopolymers has
sparked the imagination of formulation scientists because of their rec-
ognized biodegradability, biocompatibility, ease of fabrication, and sus-
tainable character [129]. Natural polymers are being exploited to their
use in the fabrication of MNs including silk fibroin [28,206], Bletilla stri-
ata polysaccharide [132], and fish scale biopolymer [60,162–164]. The
whole humankind is a spectator of the havoc that coronavirus has
caused. Recently, biodegradable MNs composed of CMC were used to
fabricate the recombinant coronavirus vaccine. It was found that signif-
icantly high SARS-CoV-2 IgG responses were detected as early as week 2
for all subunit vaccines compared to control groups [24]. Considering
the applications of biodegradable MNs in vaccination [22,23,144], it
can only be said that the future of vaccination lies in MN-assisted deliv-
ery systems.
Owing to the wide range of functional groups present in biopoly-
mers, they allow the fabrication of MNs with tunable functionalities
and properties. Thus, these biopolymer-based MNs offer great potential
not only in drug administration but also in biosensing and physiological
stimuli-responsive systems. The future of MNs lies in the use of combi-
natorial strategies. The combination of MNs and external stimuli such as
ultrasound and techniques like iontophoresis has enhanced transder-
mal drug delivery [207,208]. The research in the field of MN-based sen-
sor devices will grow rapidly in the next few years. Novel strategies of
MN drug-eluting balloon for enhanced delivery of drug to the vascular
tissue are being developed and can cause a stir in MN delivery systems
in the future [209]. Smart, wearable theranostic MN devices can be used
for personalized medication delivery. These non-invasive, cost-effective
devices will facilitate the dynamic dosing of therapeutics and bring
about a paradigm shift in the management of chronic diseases such as
cancer, diabetes, Alzheimer's disease, and infectious diseases in the fu-
ture [210].
Since the inception of 3D printing or additive manufacturing in the
1980s, it has revolutionized the pharmaceutical and biomedical fields.
3D printing-assisted fabrication of biodegradable MNs has opened a

616
N. Dabholkar, S. Gorantla, T. Waghule et al.
novel avenue for enhanced drug delivery. These devices will revolution-
ize macromolecular delivery, especially that of insulin in the future
[211,212]. Adoption of technologies such as stereolithography, fused
deposition modeling, and two-photon polymerization, will enable to
regulate MNs geometry without compromising the resolution and the
aspect ratio. Moreover, 3D printing can facilitate facile customization
of TDDS to accommodate factors such as differences in skin hydration
and thickness, that influence drug delivery [213]. Recently, a 4D printed
MN array with backward-facing barbs has been reported by Han et al. It
was demonstrated that barbed MN arrays enhanced tissue adhesion by
18 folds compared to barbless MN arrays. Enhanced tissue adhesion re-
sults in a more stable and robust performance for drug delivery, bio-
sensing, and biofluid collection. Han et al. fabricated the MNs using
poly(ethylene glycol) diacrylate. The use of biopolymers in this regard
should be investigated in the future [214].
9. Conclusion
Biodegradable and dissolving microneedles have gained significant
attraction in the field of transdermal drug delivery and have been
researched extensively by formulation scientists because of their recog-
nized biodegradability, biocompatibility, faster onset of action, ease of
fabrication, and sustainable character. Apart from the various natural
polymers used to fabricate biodegradable MNs like alginate, cellulose
and its derivatives, chitosan, chondroitin sulfate, hyaluronic acid, sugars,
and xanthan gum, polymers like silk fibroin and fish scale biopolymers
are some of the newest additions to these natural polymers. Patents in-
volving the use of natural polymers for MN fabrication are few. How-
ever, biodegradable MNs have gained entry into the preclinical and
clinical stages. With the advent of 3D printing, smart, wearable
theranostics, stimuli-responsive delivery systems, and biosensing de-
vices, MN technology holds enormous potential for clinical application
and commercialization. The future appears to be very bright for novel
delivery and monitoring systems based on MN technologies. However,
the use of natural polymers in this regard must be investigated further
to utilize the advantages associated with their biodegradability and
biocompatibility.
Declaration of competing interest
The authors declared no conflicts of interest.
References
[1] P. Mukhopadhyay, K. Sarkar, M. Chakraborty, S. Bhattacharya, R. Mishra, P.P.
Kundu, Oral insulin delivery by self-assembled chitosan nanoparticles: in vitro
and in vivo studies in diabetic animal model, Mater. Sci. Eng. C. (2013)https://
doi.org/10.1016/j.msec.2012.09.001.
[2] D.K. Mishra, V. Dhote, P.K. Mishra, Transdermal immunization: biological frame-
work and translational perspectives, Expert Opin. Drug Deliv. (2013)https://doi.
org/10.1517/17425247.2013.746660.
[3] H. Tanwar, R. Sachdeva, Transdermal drug delivery system: a review | International
Journal of Pharmaceutical Sciences and Research, Int. J. Pharm. Sci. Res. 7 (2016)
2274–90. doi:10.13040/IJPSR.0975-8232.7(6).2274-90.
[4] MARKETSANDMARKETS, Transdermal Drug Delivery System Market by Type
(Patches and Semisolid Formulations), Applications (Pain Management, Central
Nervous System Disorders, Hormonal Applications, Cardiovascular Diseases), End
User, and Region - Global Forecast to 2023, 2020.
[5] M.B. Brown, G.P. Martin, S.A. Jones, F.K. Akomeah, Dermal and transdermal drug
delivery systems: current and future prospects, Drug Deliv. 13 (2006) 175–187,
https://doi.org/10.1080/10717540500455975.
[6] D.K. Mishra, V. Pandey, R. Maheshwari, P. Ghode, R.K. Tekade, Cutaneous and
Transdermal Drug Delivery: Techniques and Delivery Systems (2018)https://doi.
org/10.1016/B978-0-12-817909-3.00015-7.
[7] T. Waghule, G. Singhvi, S.K. Dubey, M.M. Pandey, G. Gupta, M. Singh, K. Dua,
Microneedles: a smart approach and increasing potential for transdermal drug de-
livery system, Biomed. Pharmacother. 109 (2019) 1249–1258, https://doi.org/10.
1016/j.biopha.2018.10.078.
[8] M.S. Giri Nandagopal, R. Antony, S. Rangabhashiyam, N. Sreekumar, N. Selvaraju,
Overview of microneedle system: a third generation transdermal drug delivery ap-
proach, Microsyst. Technol. (2014)https://doi.org/10.1007/s00542-014-2233-5.
617
International Journal of Biological Macromolecules 170 (2021) 602–621
[9] X. He, J. Sun, J. Zhuang, H. Xu, Y. Liu, D. Wu, Microneedle system for transdermal
drug and vaccine delivery: devices, safety, and prospects, Dose-Response,
2019https://doi.org/10.1177/1559325819878585.
[10] C.J. Martin, C.J. Allender, K.R. Brain, A. Morrissey, J.C. Birchall, Low temperature fab-
rication of biodegradable sugar glass microneedles for transdermal drug delivery
applications, J. Control. Release (2012)https://doi.org/10.1016/j.jconrel.2011.10.
024.
[11] M. Sharma, Transdermal and intravenous nano drug delivery systems, in: Appl,
Target. Nano Drugs Deliv. Syst. 2019https://doi.org/10.1016/b978-0-12-814029-
1.00018-1.
[12] A.H. Sabri, Y. Kim, M. Marlow, D.J. Scurr, J. Segal, A.K. Banga, L. Kagan, J.B. Lee, Intra-
dermal and transdermal drug delivery using microneedles – fabrication, perfor-
mance evaluation and application to lymphatic delivery, Adv. Drug Deliv. Rev.
(2019)https://doi.org/10.1016/j.addr.2019.10.004.
[13] C.H. Chen, V.B.H. Shyu, C.T. Chen, Dissolving microneedle patches for transdermal
insulin delivery in diabetic mice: potential for clinical applications, Materials
(Basel), 2018https://doi.org/10.3390/ma11091625.
[14] D.P. Wermeling, S.L. Banks, D.A. Hudson, H.S. Gill, J. Gupta, M.R. Prausnitz, A.L.
Stinchcomb, Microneedles permit transdermal delivery of a skin-impermeant
medication to humans, Proc. Natl. Acad. Sci. U. S. A. (2008)https://doi.org/10.
1073/pnas.0710355105.
[15] K. Cheung, G. West, D.B. Das, Delivery of large molecular protein using flat and
short microneedles prepared using focused ion beam (FIB) as a skin ablation
tool, Drug Deliv. Transl, Res, 2015https://doi.org/10.1007/s13346-015-0252-0.
[16] T. Han, D.B. Das, Permeability enhancement for transdermal delivery of large mol-
ecule using low-frequency sonophoresis combined with microneedles, J. Pharm.
Sci. (2013)https://doi.org/10.1002/jps.23662.
[17] R.F. Donnelly, T.R.R. Singh, M.M. Tunney, D.I.J. Morrow, P.A. McCarron, C.
O’Mahony, A.D. Woolfson, Microneedle arrays allow lower microbial penetration
than hypodermic needles in vitro, Pharm. Res. (2009)https://doi.org/10.1007/
s11095-009-9967-2.
[18] B. Gualeni, S.A. Coulman, D. Shah, P.F. Eng, H. Ashraf, P. Vescovo, G.J. Blayney, L.D.
Piveteau, O.J. Guy, J.C. Birchall, Minimally invasive and targeted therapeutic cell de-
livery to the skin using microneedle devices, Br. J. Dermatol. (2018)https://doi.org/
10.1111/bjd.15923.
[19] A. Tröls, M.A. Hintermüller, M. Saeedipour, S. Pirker, B. Jakoby, Drug dosage for
microneedle-based transdermal drug delivery systems utilizing evaporation-
induced droplet transport, Microfluid, Nanofluidics, 2019https://doi.org/10.1007/
s10404-019-2257-3.
[20] Z. Zhao, Y. Chen, Y, Shi, Microneedles, a potential strategy in transdermal delivery
and application in the management of psoriasis, RSC Adv, 2020https://doi.org/10.
1039/d0ra00735h.
[21] S.H. Bariya, M.C. Gohel, T.A. Mehta, O.P. Sharma, Microneedles: an emerging trans-
dermal drug delivery system, J. Pharm. Pharmacol. (2012)https://doi.org/10.1111/
j.2042-7158.2011.01369.x.
[22] M.J. Mistilis, A.S. Bommarius, M.R. Prausnitz, Development of a thermostable
microneedle patch for influenza vaccination, J. Pharm. Sci. (2015)https://doi.org/
10.1002/jps.24283.
[23] E. Kim, G. Erdos, S. Huang, T. Kenniston, L.D. Falo, A. Gambotto, Preventative vac-
cines for zika virus outbreak: preliminary evaluation, EBioMedicine, 2016https://
doi.org/10.1016/j.ebiom.2016.09.028.
[24] E. Kim, G. Erdos, S. Huang, T.W. Kenniston, S.C. Balmert, C.D. Carey, V.S. Raj, M.W.
Epperly, W.B. Klimstra, B.L. Haagmans, E. Korkmaz, L.D. Falo, A. Gambotto,
Microneedle array delivered recombinant coronavirus vaccines: immunogenicity
and rapid translational development, EBioMedicine, 2020https://doi.org/10.1016/
j.ebiom.2020.102743.
[25] K. Fukushima, T. Yamazaki, R. Hasegawa, Y. Ito, N. Sugioka, K. Takada, Pharmacoki-
netic and pharmacodynamic evaluation of insulin dissolving microneedles in dogs.,
Diabetes Technol. Ther. (2010). doi:https://doi.org/10.1089/dia.2009.0176.
[26] S. Liu, M.N. Jin, Y.S. Quan, F. Kamiyama, H. Katsumi, T. Sakane, A. Yamamoto, The
development and characteristics of novel microneedle arrays fabricated from
hyaluronic acid, and their application in the transdermal delivery of insulin, J. Con-
trol. Release (2012)https://doi.org/10.1016/j.jconrel.2012.05.030.
[27] J.W. Lee, S.O. Choi, E.I. Felner, M.R. Prausnitz, Dissolving microneedle patch for
transdermal delivery of human growth hormone, Small, 2011https://doi.org/10.
1002/smll.201001091.
[28] K. Tsioris, W.K. Raja, E.M. Pritchard, B. Panilaitis, D.L. Kaplan, F.G. Omenetto, Fabri-
cation of silk microneedles for controlled-release drug delivery, Adv. Funct. Mater.
(2012)https://doi.org/10.1002/adfm.201102012.
[29] W. Li, R.N. Terry, J. Tang, M.R. Feng, S.P. Schwendeman, M.R. Prausnitz, Rapidly sep-
arable microneedle patch for the sustained release of a contraceptive, Nat. Biomed,
Eng, 2019https://doi.org/10.1038/s41551-018-0337-4.
[30] S. Bhatnagar, P. Kumari, S.P. Pattarabhiran, V.V.K. Venuganti, Zein microneedles for
localized delivery of chemotherapeutic agents to treat breast cancer: drug loading,
release behavior, and skin permeation studies, AAPS PharmSciTech, 2018https://
doi.org/10.1208/s12249-018-1004-5.
[31] Y. Ito, H. Murano, N. Hamasaki, K. Fukushima, K. Takada, Incidence of low bioavail-
ability of leuprolide acetate after percutaneous administration to rats by dissolving
microneedles, Int. J. Pharm. (2011)https://doi.org/10.1016/j.ijpharm.2011.01.039.
[32] L. Xie, H. Zeng, J. Sun, W. Qian, Engineering microneedles for therapy and diagno-
sis: a survey, Micromachines, 2020https://doi.org/10.3390/mi11030271.
[33] A.Z. Alkilani, M.T.C. McCrudden, R.F. Donnelly, Transdermal drug delivery: Innova-
tive pharmaceutical developments based on disruption of the barrier properties of
the stratum corneum, Pharmaceutics, 2015https://doi.org/10.3390/
pharmaceutics7040438.
N. Dabholkar, S. Gorantla, T. Waghule et al.
[34] Y.B. Schuetz, A. Naik, R.H. Guy, Y.N. Kalia, Emerging strategies for the transdermal
delivery of peptide and protein drugs, Expert Opin. Drug Deliv. (2005)https://doi.
org/10.1517/17425247.2.3.533.
[35] C.M. Schoellhammer, D. Blankschtein, R. Langer, Skin permeabilization for trans-
dermal drug delivery: recent advances and future prospects, Expert Opin. Drug
Deliv. (2014)https://doi.org/10.1517/17425247.2014.875528.
[36] Y. Shahzad, R. Louw, M. Gerber, J. Du Plessis, Breaching the skin barrier through
temperature modulations, J. Control. Release (2015)https://doi.org/10.1016/j.
jconrel.2015.01.019.
[37] S. Dharadhar, A. Majumdar, S. Dhoble, V. Patravale, Microneedles for transdermal
drug delivery: a systematic review, Drug Dev. Ind. Pharm. (2019)https://doi.org/
10.1080/03639045.2018.1539497.
[38] M. Pearton, V. Saller, S.A. Coulman, C. Gateley, A.V. Anstey, V. Zarnitsyn, J.C. Birchall,
Microneedle delivery of plasmid DNA to living human skin: formulation coating,
skin insertion and gene expression, J. Control. Release 160 (2012) 561–569,
https://doi.org/10.1016/j.jconrel.2012.04.005.
[39] M. Sharma, Transdermal and intravenous nano drug delivery systems, in: Appl,
Elsevier, Target. Nano Drugs Deliv. Syst., 2019 499–550, https://doi.org/10.1016/
b978-0-12-814029-1.00018-1.
[40] E. Larrañeta, R.E.M. Lutton, A.D. Woolfson, R.F. Donnelly, Microneedle arrays as
transdermal and intradermal drug delivery systems: materials science, manufac-
ture and commercial development, Mater. Sci. Eng. R Reports. (2016)https://doi.
org/10.1016/j.mser.2016.03.001.
[41] K.J. Lee, S.S. Jeong, D.H. Roh, D.Y. Kim, H.K. Choi, E.H. Lee, A practical guide to the
development of microneedle systems – in clinical trials or on the market, Int. J.
Pharm. (2020)https://doi.org/10.1016/j.ijpharm.2019.118778.
[42] L. Wei-Ze, H. Mei-Rong, Z. Jian-Ping, Z. Yong-Qiang, H. Bao-Hua, L. Ting, Z. Yong,
Super-short solid silicon microneedles for transdermal drug delivery applications,
Int. J. Pharm. (2010)https://doi.org/10.1016/j.ijpharm.2010.01.024.
[43] S. Pradeep Narayanan, S. Raghavan, Fabrication and characterization of gold-coated
solid silicon microneedles with improved biocompatibility, Int. J. Adv. Manuf.
Technol. (2019)https://doi.org/10.1007/s00170-018-2596-3.
[44] W. Chen, H. Li, D. Shi, Z. Liu, W. Yuan, Microneedles as a delivery system for gene
therapy, Front, Pharmacol, 2016https://doi.org/10.3389/fphar.2016.00137.
[45] S. Doddaballapur, Microneedling with dermaroller, J. Cutan. Aesthet, Surg,
2009https://doi.org/10.4103/0974-2077.58529.
[46] Y. Kapoor, M. Milewski, L. Dick, J. Zhang, J.R. Bothe, M. Gehrt, K. Manser, B. Nissley,
I. Petrescu, P. Johnson, S. Burton, J. Moseman, V. Hua, T. Grunewald, M. Tomai, R.
Smith, Coated microneedles for transdermal delivery of a potent pharmaceutical
peptide, Biomed. Microdevices (2020)https://doi.org/10.1007/s10544-019-0462-
1.
[47] S. Li, W. Li, M. Prausnitz, Individually coated microneedles for co-delivery of multi-
ple compounds with different properties, Drug Deliv. Transl, Res, 2018https://doi.
org/10.1007/s13346-018-0549-x.
[48] J.H. Park, M.G. Allen, M.R. Prausnitz, Polymer microneedles for controlled-release
drug delivery, Pharm. Res. (2006)https://doi.org/10.1007/s11095-006-0028-9.
[49] M. Wang, L. Hu, C. Xu, Recent advances in the design of polymeric microneedles for
transdermal drug delivery and biosensing, Lab Chip, 2017https://doi.org/10.1039/
C7LC00016B.
[50] S. Fakhraei Lahiji, Y. Kim, G. Kang, S. Kim, S. Lee, H. Jung, Tissue interlocking dissolv-
ing microneedles for accurate and efficient transdermal delivery of biomolecules,
Sci. Rep. (2019)https://doi.org/10.1038/s41598-019-44418-6.
[51] S. Lee, S.F. Lahiji, J. Jang, M. Jang, H. Jung, Micro-pillar integrated dissolving
microneedles for enhanced transdermal drug delivery, Pharmaceutics,
2019https://doi.org/10.3390/pharmaceutics11080402.
[52] M.H. Ling, M.C. Chen, Dissolving polymer microneedle patches for rapid and effi-
cient transdermal delivery of insulin to diabetic rats, Acta Biomater. 9 (2013)
8952–8961, https://doi.org/10.1016/j.actbio.2013.06.029.
[53] J.J. Norman, S.O. Choi, N.T. Tong, A.R. Aiyar, S.R. Patel, M.R. Prausnitz, M.G. Allen,
Hollow microneedles for intradermal injection fabricated by sacrificial
micromolding and selective electrodeposition, Biomed. Microdevices (2013)
https://doi.org/10.1007/s10544-012-9717-9.
[54] R. Mishra, T.K. Maiti, T.K. Bhattacharyya, Development of SU-8 hollow
microneedles on a silicon substrate with microfluidic interconnects for transder-
mal drug delivery, J. Micromechanics Microengineering. (2018)https://doi.org/
10.1088/1361-6439/aad301.
[55] X. Hong, L. Wei, F. Wu, Z. Wu, L. Chen, Z. Liu, W. Yuan, Dissolving and biodegrad-
able microneedle technologies for transdermal sustained delivery of drug and vac-
cine, Drug Des. Devel, Ther, 2013https://doi.org/10.2147/DDDT.S44401.
[56] J.H. Park, M.G. Allen, M.R. Prausnitz, Biodegradable polymer microneedles: fabrica-
tion, mechanics and transdermal drug delivery, J. Control. Release (2005)https://
doi.org/10.1016/j.jconrel.2005.02.002.
[57] Q.Y. Li, J.N. Zhang, B.Z. Chen, Q.L. Wang, X.D. Guo, A solid polymer microneedle
patch pretreatment enhances the permeation of drug molecules into the skin,
RSC Adv, 2017https://doi.org/10.1039/c6ra26759a.
[58] S.C. Park, M.J. Kim, S.K. Baek, J.H. Park, S.O. Choi, Spray-formed layered polymer
microneedles for controlled biphasic drug delivery, Polymers (Basel),
2019https://doi.org/10.3390/POLYM11020369.
[59] Z. Luo, W. Sun, J. Fang, K.J. Lee, S. Li, Z. Gu, M.R. Dokmeci, A. Khademhosseini, Bio-
degradable gelatin methacryloyl microneedles for transdermal drug delivery, Adv.
Healthc, Mater, 2019https://doi.org/10.1002/adhm.201801054.
[60] P. Medhi, O. Olatunji, A. Nayak, C.T. Uppuluri, R.T. Olsson, B.N. Nalluri, D.B. Das,
Lidocaine-loaded fish scale-nanocellulose biopolymer composite microneedles,
AAPS PharmSciTech, 2017https://doi.org/10.1208/s12249-017-0758-5.
618
International Journal of Biological Macromolecules 170 (2021) 602–621
[61] Y. Cao, Y. Tao, Y. Zhou, S. Gui, Development of sinomenine hydrochloride-loaded
polyvinylalcohol/maltose microneedle for transdermal delivery, J. Drug Deliv. Sci.
Technol. (2016)https://doi.org/10.1016/j.jddst.2016.06.007.
[62] M.B. Perez Cuevas, M. Kodani, Y. Choi, J. Joyce, S.M. O’Connor, S. Kamili, M.R.
Prausnitz, Hepatitis B vaccination using a dissolvable microneedle patch is immu-
nogenic in mice and rhesus macaques, Bioeng. Transl, Med, 2018https://doi.org/
10.1002/btm2.10098.
[63] Y.C. Kim, F.S. Quan, J.M. Song, A. Vunnava, D.G. Yoo, K.M. Park, R.W. Compans, S.M.
Kang, M.R. Prausnitz, Influenza immunization with trehalose-stabilized virus-like
particle vaccine using microneedles, Procedia Vaccinol, 2010https://doi.org/10.
1016/j.provac.2010.03.004.
[64] G. Cole, A.A. Ali, C.M. McCrudden, J.W. McBride, J. McCaffrey, T. Robson, V.L. Kett,
N.J. Dunne, R.F. Donnelly, H.O. McCarthy, DNA vaccination for cervical cancer: stra-
tegic optimisation of RALA mediated gene delivery from a biodegradable
microneedle system, Eur. J. Pharm. Biopharm. (2018)https://doi.org/10.1016/j.
ejpb.2018.02.029.
[65] M. He, G. Yang, X. Zhao, S. Zhang, Y. Gao, Intradermal implantable PLGA
microneedles for etonogestrel sustained release, J. Pharm. Sci. (2020)https://doi.
org/10.1016/j.xphs.2020.02.009.
[66] M. Battisti, R. Vecchione, C. Casale, F.A. Pennacchio, V. Lettera, R. Jamaledin, M.
Profeta, C. Di Natale, G. Imparato, F. Urciuolo, P.A. Netti, Non-invasive production
of multi-compartmental biodegradable polymer microneedles for controlled intra-
dermal drug release of labile molecules, Front. Bioeng, Biotechnol, 2019https://doi.
org/10.3389/fbioe.2019.00296.
[67] E. Marin, M.I. Briceño, C. Caballero-George, Critical evaluation of biodegradable
polymers used in nanodrugs, Int. J. Nanomedicine 8 (2013) 3071–3091, https://
doi.org/10.2147/IJN.S47186.
[68] N.R. Nair, V.C. Sekhar, K.M. Nampoothiri, A. Pandey, Biodegradation of biopoly-
mers, in: Curr, Elsevier Inc., Dev. Biotechnol. Bioeng. Prod. Isol. Purif. Ind. Prod,
2016 739–755, https://doi.org/10.1016/B978-0-444-63662-1.00032-4.
[69] K.I. Draget, Alginates, in: Handb, Hydrocoll. Second Ed. 2009https://doi.org/10.
1533/9781845695873.807.
[70] A.R. Nesic, S.I. Seslija, The influence of nanofillers on physical–chemical properties
of polysaccharide-based film intended for food packaging, Food Packag,
2017https://doi.org/10.1016/b978-0-12-804302-8.00019-4.
[71] C.H. Goh, P.W.S. Heng, L.W. Chan, Alginates as a useful natural polymer for micro-
encapsulation and therapeutic applications, Carbohydr. Polym. (2012)https://doi.
org/10.1016/j.carbpol.2011.11.012.
[72] R.D. Koyani, Biopolymers for microneedle synthesis: from then to now,
Biomanufacturing Rev, 2019https://doi.org/10.1007/s40898-019-0006-8.
[73] T. Heinze, O.A. El Seoud, A. Koschella, Production and Characteristics of Cellulose
From Different Sources, in, 2018https://doi.org/10.1007/978-3-319-73168-1_1.
[74] J. Shokri, K. Adibki, Application of cellulose and cellulose derivatives in pharmaceu-
tical industries, in: Cellul. - Medical, Pharm. Electron. Appl., 2013. doi:https://doi.
org/10.5772/55178.
[75] J.E. Song, S.H. Jun, S.G. Park, N.G. Kang, A semi-dissolving microneedle patch incor-
porating TEMPO-oxidized bacterial cellulose nanofibers for enhanced transdermal
delivery, Polymers (Basel). 12 (2020) 1873. doi:https://doi.org/10.3390/
POLYM12091873.
[76] D.I. Sánchez-Machado, J. López-Cervantes, M.A. Correa-Murrieta, R.G. Sánchez-
Duarte, P. Cruz-Flores, G.S. la Mora-López, Chitosan, in: Nonvitamin Nonmineral
Nutr, Suppl. 2018https://doi.org/10.1016/B978-0-12-812491-8.00064-3.
[77] F. Shahidi, R. Abuzaytoun, Chitin, chitosan, and co-products : chemistry, produc-
tion, applications, and health effects, 49 (2005).
[78] J. Lizardi-Mendoza, W.M. Argüelles Monal, F.M. Goycoolea Valencia, Chemical
characteristics and functional properties of chitosan, in: Chitosan Preserv, Agric.
Commod. 2016https://doi.org/10.1016/B978-0-12-802735-6.00001-X.
[79] J. Chi, X. Zhang, C. Chen, C. Shao, Y. Zhao, Y. Wang, Antibacterial and angiogenic chi-
tosan microneedle array patch for promoting wound healing, Bioact. Mater. 5
(2020) 253–259, https://doi.org/10.1016/j.bioactmat.2020.02.004.
[80] M. Nurunnabi, V. Revuri, K.M. Huh, Y. kyu Lee, Polysaccharide based nano/
microformulation: an effective and versatile oral drug delivery system, in: Nano-
structures Oral Med., 2017. doi:https://doi.org/10.1016/B978-0-323-47720-8.
00015-8.
[81] F.G. Huffman, Uronic acids, in: Encycl, Food Sci. Nutr. 2003https://doi.org/10.1016/
b0-12-227055-x/01221-9.
[82] A. Schieber, D. Lopes-Lutz, Analytical methods - functional foods and dietary sup-
plements, in: Compr, Biotechnol. Second Ed. 2011https://doi.org/10.1016/B978-
0-08-088504-9.00320-2.
[83] C.G. Boeriu, J. Springer, F.K. Kooy, L.A.M. van den Broek, G. Eggink, Production
methods for hyaluronan, Int. J. Carbohydr, Chem, 2013https://doi.org/10.1155/
2013/624967.
[84] J. Zhu, X. Tang, Y. Jia, C.T. Ho, Q. Huang, Applications and delivery mechanisms of
hyaluronic acid used for topical/transdermal delivery – a review, Int. J. Pharm.
578 (2020) 119127, https://doi.org/10.1016/j.ijpharm.2020.119127.
[85] J.U. Shin, J.D. Kim, H.K. Kim, H.K. Kang, C. Joo, J.H. Lee, D.H. Jeong, S. Song, H. Chu, J.S.
Lee, H. Lee, K.H. Lee, The use of biodegradable microneedle patches to increase
penetration of topical steroid for prurigo nodularis, Eur. J, Dermatology,
2018https://doi.org/10.1684/ejd.2017.3164.
[86] Y. Kim, S.A. Bhattaccharjee, M. Beck-Broichsitter, A.K. Banga, Fabrication and char-
acterization of hyaluronic acid microneedles to enhance delivery of magnesium
ascorbyl phosphate into skin, Biomed. Microdevices (2019)https://doi.org/10.
1007/s10544-019-0455-0.
[87] G. Bonfante, H. Lee, L. Bao, J. Park, N. Takama, B. Kim, Comparison of polymers to
enhance mechanical properties of microneedles for bio-medical applications,
Micro Nano Syst. Lett. 8 (2020) 13, https://doi.org/10.1186/s40486-020-00113-0.
N. Dabholkar, S. Gorantla, T. Waghule et al.
[88] M. Zaitoun, M. Ghanem, S. Harphoush, Sugars : types and their functional proper-
ties in food and human health, Int. J. Public Heal. Res. 6 (4) (2019) 93–99.
[89] P. Serrano-Castañeda, J.J. Escobar-Chávez, I.M. Rodríguez-Cruz, L.M. Melgoza-
Contreras, J. Martínez-Hernández, Microneedles as enhancer of drug absorption
through the skin and applications in medicine and cosmetology, J. Pharm.
Pharm. Sci. (2018). doi:10.18433/jpps29610.
[90] R.F. Donnelly, D.I.J. Morrow, T.R.R. Singh, K. Migalska, P.A. McCarron, C. O’Mahony,
A.D. Woolfson, Processing difficulties and instability of carbohydrate microneedle
arrays, Drug Dev. Ind. Pharm. (2009)https://doi.org/10.1080/
03639040902882280.
[91] E.P. Yalcintas, D.S. Ackerman, E. Korkmaz, C.A. Telmer, J.W. Jarvik, P.G. Campbell,
M.P. Bruchez, O.B. Ozdoganlar, Analysis of in vitro cytotoxicity of carbohydrate-
based materials used for dissolvable microneedle arrays, Pharm. Res. (2020)
https://doi.org/10.1007/s11095-019-2748-7.
[92] E. Larrañeta, R.E.M. Lutton, A.D. Woolfson, R.F. Donnelly, Microneedle arrays as
transdermal and intradermal drug delivery systems: materials science, manufac-
ture and commercial development, Mater. Sci. Eng. R Reports. 104 (2016) 1–32,
https://doi.org/10.1016/j.mser.2016.03.001.
[93] G. Singhvi, N. Hans, N. Shiva, S. Kumar Dubey, Xanthan gum in drug delivery appli-
cations, in: Nat, Elsevier, Polysaccharides Drug Deliv. Biomed. Appl., 2019 121–144,
https://doi.org/10.1016/b978-0-12-817055-7.00005-4.
[94] G. Sworn, Xanthan gum, in: Handb, Hydrocoll. Second Ed. 2009https://doi.org/10.
1533/9781845695873.186.
[95] R.S.J. Ingrole, H.S. Gill, Microneedle coating methods: a review with a perspective, J.
Pharmacol. Exp. Ther. (2019)https://doi.org/10.1124/jpet.119.258707.
[96] Y.K. Demir, Z. Akan, O. Kerimoglu, Characterization of polymeric microneedle ar-
rays for transdermal drug delivery, PLoS One, 2013https://doi.org/10.1371/
journal.pone.0077289.
[97] Y.K. Demir, Z. Akan, O. Kerimoglu, Sodium alginate microneedle arrays mediate the
transdermal delivery of bovine serum albumin, PLoS One, 2013https://doi.org/10.
1371/journal.pone.0063819.
[98] Y. Zhang, G. Jiang, W. Yu, D. Liu, B. Xu, Microneedles fabricated from alginate and
maltose for transdermal delivery of insulin on diabetic rats, Mater. Sci. Eng. C.
(2018)https://doi.org/10.1016/j.msec.2017.12.006.
[99] F. Dias, C. Duarte, Cellulose and its derivatives use in the pharmaceutical
compounding practice, Cellul. - Medical, Pharm. Electron. Appl. (2013). doi:
https://doi.org/10.5772/56637.
[100] R. Ergun, J. Guo, B. Huebner-Keese, Cellulose, in: Encycl, Food Heal. 2015https://
doi.org/10.1016/B978-0-12-384947-2.00127-6.
[101] Y.H. Park, S.K. Ha, I. Choi, K.S. Kim, J. Park, N. Choi, B. Kim, J.H. Sung, Fabrication of
degradable carboxymethyl cellulose (CMC) microneedle with laser writing and
replica molding process for enhancement of transdermal drug delivery, Biotechnol.
Bioprocess Eng. (2016)https://doi.org/10.1007/s12257-015-0634-7.
[102] L.E. González García, M.N. MacGregor, R.M. Visalakshan, N. Ninan, A.A. Cavallaro,
A.D. Trinidad, Y. Zhao, A.J.D. Hayball, K. Vasilev, Self-sterilizing antibacterial
silver-loaded microneedles, Chem. Commun. (2019)https://doi.org/10.1039/
c8cc06035e.
[103] M.C. Chen, S.F. Huang, K.Y. Lai, M.H. Ling, Fully embeddable chitosan microneedles
as a sustained release depot for intradermal vaccination, Biomaterials, 2013https://
doi.org/10.1016/j.biomaterials.2012.12.041.
[104] Z. Ahmad, M.I. Khan, M.I. Siddique, H.S. Sarwar, G. Shahnaz, S.Z. Hussain, N.I.
Bukhari, I. Hussain, M.F. Sohail, Fabrication and characterization of thiolated chito-
san microneedle patch for transdermal delivery of tacrolimus, AAPS PharmSciTech,
2020https://doi.org/10.1208/s12249-019-1611-9.
[105] N. ‘Izzah Ibrahim, S.K. Wong, I.N. Mohamed, N. Mohamed, K.Y. Chin, S. Ima-
Nirwana, A.N. Shuid, Wound healing properties of selected natural products, Int.
J. Environ. Res. Public Health. 15 (2018). doi:https://doi.org/10.3390/
ijerph15112360.
[106] M.C. Chen, M.H. Ling, K.Y. Lai, E. Pramudityo, Chitosan microneedle patches for
sustained transdermal delivery of macromolecules, Biomacromolecules,
2012https://doi.org/10.1021/bm301293d.
[107] A. Sadeqi, H.R. Nejad, G. Kiaee, S. Sonkusale, Cost-effective fabrication of chitosan
microneedles for transdermal drug delivery, Conf. Proc. ... Annu. Int. Conf. IEEE
Eng. Med. Biol. Soc. IEEE Eng. Med. Biol. Soc. Annu. Conf. (2018). doi:https://doi.
org/10.1109/EMBC.2018.8513691.
[108] A. Chandrasekharan, Y.J. Hwang, K.Y. Seong, S. Park, S. Kim, S.Y. Yang, Acid-treated
water-soluble chitosan suitable for microneedle-assisted intracutaneous drug de-
livery, Pharmaceutics, 2019https://doi.org/10.3390/pharmaceutics11050209.
[109] J. Chi, X. Zhang, C. Chen, C. Shao, Y. Zhao, Y. Wang, Antibacterial and angiogenic chi-
tosan microneedle array patch for promoting wound healing, Bioact, Mater,
2020https://doi.org/10.1016/j.bioactmat.2020.02.004.
[110] Y. gang Shi, Y. cheng Meng, J. rong Li, J. Chen, Y. hua Liu, X. Bai, Chondroitin sulfate:
extraction, purification, microbial and chemical synthesis, J. Chem. Technol.
Biotechnol. (2014). doi:https://doi.org/10.1002/jctb.4454.
[111] A. Aravamudhan, D.M. Ramos, A.A. Nada, S.G. Kumbar, Natural polymers, in: Nat,
Synth. Biomed. Polym. 2014https://doi.org/10.1016/b978-0-12-396983-5.00004-
1.
[112] D. Poirier, F. Renaud, V. Dewar, L. Strodiot, F. Wauters, J. Janimak, T. Shimada, T.
Nomura, K. Kabata, K. Kuruma, T. Kusano, M. Sakai, H. Nagasaki, T. Oyamada, Hep-
atitis B surface antigen incorporated in dissolvable microneedle array patch is an-
tigenic and thermostable, Biomaterials, 2017https://doi.org/10.1016/j.
biomaterials.2017.08.038.
[113] S. Liu, S. Zhang, Y. Duan, Y. Niu, H. Gu, Z. Zhao, S. Zhang, Y. Yang, X. Wang, Y. Gao, P.
Yang, Transcutaneous immunization of recombinant Staphylococcal enterotoxin B
protein using a dissolving microneedle provides potent protection against lethal
619
International Journal of Biological Macromolecules 170 (2021) 602–621
enterotoxin challenge, Vaccine, 2019https://doi.org/10.1016/j.vaccine.2019.05.
055.
[114] P. Snetkov, K. Zakharova, S. Morozkina, R. Olekhnovich, M. Uspenskaya, Hyaluronic
acid: the influence of molecular weight on structural, physical, physico-chemical,
and degradable properties of biopolymer, Polymers (Basel). 12 (2020). doi:
https://doi.org/10.3390/polym12081800.
[115] S. Vasvani, P. Kulkarni, D. Rawtani, Hyaluronic acid: a review on its biology, aspects
of drug delivery, route of administrations and a special emphasis on its approved
marketed products and recent clinical studies, Int. J. Biol. Macromol. (2019)
https://doi.org/10.1016/j.ijbiomac.2019.11.066.
[116] J.R.E. Fraser, T.C. Laurent, U.B.G. Laurent, Hyaluronan: its nature, distribution, func-
tions and turnover, in: J, Intern. Med. (1997)https://doi.org/10.1046/j.1365-2796.
1997.00170.x.
[117] J.H. Sze, J.C. Brownlie, C.A. Love, Biotechnological production of hyaluronic acid: a
mini review, 3 Biotech, 2016https://doi.org/10.1007/s13205-016-0379-9.
[118] S.G. Lee, J.H. Jeong, K.M. Lee, K.H. Jeong, H. Yang, M. Kim, H. Jung, S. Lee, Y.W. Choi,
Nanostructured lipid carrier-loaded hyaluronic acid microneedles for controlled
dermal delivery of a lipophilic molecule, Int. J, Nanomedicine, 2013https://doi.
org/10.2147/IJN.S54529.
[119] S.Y. Choi, H.J. Kwon, G.R. Ahn, E.J. Ko, K.H. Yoo, B.J. Kim, C. Lee, D. Kim, Hyaluronic
acid microneedle patch for the improvement of crow’s feet wrinkles, Dermatol.
Ther. (2017)https://doi.org/10.1111/dth.12546.
[120] Y. Hao, Y. Chen, M. Lei, T. Zhang, Y. Cao, J. Peng, L. Chen, Z. Qian, Near-infrared re-
sponsive PEGylated gold nanorod and doxorubicin loaded dissolvable hyaluronic
acid microneedles for human epidermoid cancer therapy, Adv. Ther. (2018)
https://doi.org/10.1002/adtp.201800008.
[121] H. Yang, X. Wu, Z. Zhou, X. Chen, M. Kong, Enhanced transdermal lymphatic deliv-
ery of doxorubicin via hyaluronic acid based transfersomes/microneedle complex
for tumor metastasis therapy, Int. J. Biol. Macromol. 125 (2019) 9–16, https://doi.
org/10.1016/j.ijbiomac.2018.11.230.
[122] S. Selwyn, J. Durodie, The antimicrobial activity of sugar against pathogens of
wounds and other infections of man, in: Prop, Water Foods, Springer
Netherlands, 1985 293–308, https://doi.org/10.1007/978-94-009-5103-7_18.
[123] M. Milewski, Y. Kapoor, Z. Ding, J. Zhang, E. Ghartey-Tagoe, K. Manser, B. Nissley, I.
Petrescu, L. Xu, B. Duffield, G. Chen, Q. Yang, P. Singh, R. Smith, Stabilization and
transdermal delivery of an investigational peptide using MicroCor® solid-state dis-
solving microstructure arrays, J. Pharm. Sci. (2020)https://doi.org/10.1016/j.xphs.
2019.11.006.
[124] T. Ilić, S. Savić, B. Batinić, B. Marković, M. Schmidberger, D. Lunter, M. Savić, S. Savić,
Combined use of biocompatible nanoemulsions and solid microneedles to improve
transport of a model NSAID across the skin: in vitro and in vivo studies, Eur. J.
Pharm. Sci. (2018)https://doi.org/10.1016/j.ejps.2018.09.023.
[125] S. Gopi, A. Amalraj, N.P. Sukumaran, J.T. Haponiuk, S. Thomas, Biopolymers and
their composites for drug delivery: a brief review, Macromol. Symp. (2018)
https://doi.org/10.1002/masy.201800114.
[126] H.S. Gill, M.R. Prausnitz, Coating formulations for microneedles, Pharm. Res. (2007)
https://doi.org/10.1007/s11095-007-9286-4.
[127] Y.C. Kim, F.S. Quan, R.W. Compans, S.M. Kang, M.R. Prausnitz, Formulation of
microneedles coated with influenza virus-like particle vaccine, AAPS
PharmSciTech, 2010https://doi.org/10.1208/s12249-010-9471-3.
[128] H.J. Choi, J.M. Song, B.J. Bondy, R.W. Compans, S.M. Kang, M.R. Prausnitz, Effect of
osmotic pressure on the stability of whole inactivated influenza vaccine for coating
on microneedles, PLoS One, 2015https://doi.org/10.1371/journal.pone.0134431.
[129] D.F.S. Fonseca, P.C. Costa, I.F. Almeida, P. Dias-Pereira, I. Correia-Sá, V. Bastos, H.
Oliveira, M. Duarte-Araújo, M. Morato, C. Vilela, A.J.D. Silvestre, C.S.R. Freire,
Pullulan microneedle patches for the efficient transdermal administration of insu-
lin envisioning diabetes treatment, Carbohydr. Polym. (2020)https://doi.org/10.
1016/j.carbpol.2020.116314.
[130] L.K. Vora, A.J. Courtenay, I.A. Tekko, E. Larrañeta, R.F. Donnelly, Pullulan-based dis-
solving microneedle arrays for enhanced transdermal delivery of small and large
biomolecules, Int. J. Biol. Macromol. (2020)https://doi.org/10.1016/j.ijbiomac.
2019.12.184.
[131] S. Luo, S. Song, C. Zheng, Y. Wang, X. Xia, B. Liang, G. Feng, Biocompatibility of
Bletilla striata Microspheres as a Novel Embolic Agent (2015)https://doi.org/10.
1155/2015/840896.
[132] L. Hu, Z. Liao, Q. Hu, K.G. Maffucci, Y. Qu, Novel Bletilla striata polysaccharide
microneedles: fabrication, characterization, and in vitro transcutaneous drug de-
livery, Int. J. Biol. Macromol. (2018)https://doi.org/10.1016/j.ijbiomac.2018.05.
097.
[133] W. Yu, G. Jiang, Y. Zhang, D. Liu, B. Xu, J. Zhou, Polymer microneedles fabricated
from alginate and hyaluronate for transdermal delivery of insulin, Mater. Sci.
Eng. C. (2017)https://doi.org/10.1016/j.msec.2017.05.143.
[134] S. Kim, J. Eum, H. Yang, H. Jung, Transdermal finasteride delivery via powder-
carrying microneedles with a diffusion enhancer to treat androgenetic alopecia,
J. Control. Release (2019)https://doi.org/10.1016/j.jconrel.2019.11.002.
[135] S. Fakhraei Lahiji, S.H. Seo, S. Kim, M. Dangol, J. Shim, C.G. Li, Y. Ma, C. Lee, G. Kang,
H. Yang, K.Y. Choi, H. Jung, Transcutaneous implantation of valproic acid-
encapsulated dissolving microneedles induces hair regrowth, Biomaterials,
2018https://doi.org/10.1016/j.biomaterials.2018.03.019.
[136] H.R. Jeong, J.Y. Kim, S.N. Kim, J.H. Park, Local dermal delivery of cyclosporin a, a hy-
drophobic and high molecular weight drug, using dissolving microneedles, Eur. J.
Pharm. Biopharm. (2018)https://doi.org/10.1016/j.ejpb.2018.02.014.
[137] J.Y. Kim, M.R. Han, Y.H. Kim, S.W. Shin, S.Y. Nam, J.H. Park, Tip-loaded dissolving
microneedles for transdermal delivery of donepezil hydrochloride for treatment
of Alzheimer’s disease, Eur. J. Pharm. Biopharm. 105 (2016) 148–155, https://doi.
org/10.1016/j.ejpb.2016.06.006.
N. Dabholkar, S. Gorantla, T. Waghule et al.
[138] K. Siddhapura, H. Harde, S. Jain, Immunostimulatory effect of tetanus toxoid loaded
chitosan nanoparticles following microneedles assisted immunization,
Nanomedicine Nanotechnology, Biol. Med, 2016https://doi.org/10.1016/j.nano.
2015.10.009.
[139] Y.-S. Tsai, M.-Y. Chen, S.-K. Lan, H.-T. Tsai, M.-C. Chen, T.-S. Tzai, Transdermal deliv-
ery of leuprolide acetate with chitosan microneedles: a promising tool for andro-
gen deprivation therapy, Eur. Urol. Suppl. (2017)https://doi.org/10.1016/s1569-
9056(17)30808-4.
[140] P. Schipper, K. van der Maaden, V. Groeneveld, M. Ruigrok, S. Romeijn, S. Uleman,
C. Oomens, G. Kersten, W. Jiskoot, J. Bouwstra, Diphtheria toxoid and N-trimethyl
chitosan layer-by-layer coated pH-sensitive microneedles induce potent immune
responses upon dermal vaccination in mice, J. Control. Release (2017)https://doi.
org/10.1016/j.jconrel.2017.07.017.
[141] M.C. Chen, K.Y. Lai, M.H. Ling, C.W. Lin, Enhancing immunogenicity of antigens
through sustained intradermal delivery using chitosan microneedles with a
patch-dissolvable design, Acta Biomater, 2018https://doi.org/10.1016/j.actbio.
2017.11.004.
[142] T. Tomono, A new way to control the internal structure of microneedles: a case of
chitosan lactate, Mater, Today Chem, 2019https://doi.org/10.1016/j.mtchem.2019.
04.009.
[143] A.F. Moreira, C.F. Rodrigues, T.A. Jacinto, S.P. Miguel, E.C. Costa, I.J. Correia, Poly
(vinyl alcohol)/chitosan layer-by-layer microneedles for cancer chemo-
photothermal therapy, Int. J. Pharm. (2020)https://doi.org/10.1016/j.ijpharm.
2019.118907.
[144] A. Nakatsukasa, K. Kuruma, M. Okamatsu, T. Hiono, M. Suzuki, K. Matsuno, H. Kida,
T. Oyamada, Y. Sakoda, Potency of whole virus particle and split virion vaccines
using dissolving microneedle against challenges of H1N1 and H5N1 influenza vi-
ruses in mice, Vaccine, 2017https://doi.org/10.1016/j.vaccine.2017.04.009.
[145] W. Yao, C. Tao, J. Zou, H. Zheng, J. Zhu, Z. Zhu, J. Zhu, L. Liu, F. Li, X. Song, Flexible
two-layer dissolving and safing microneedle transdermal of neurotoxin: a
biocomfortable attempt to treat rheumatoid arthritis, Int. J. Pharm. 563 (2019)
91–100, https://doi.org/10.1016/j.ijpharm.2019.03.033.
[146] S. Naito, Y. Ito, T. Kiyohara, M. Kataoka, M. Ochiai, K. Takada, Antigen-loaded dis-
solving microneedle array as a novel tool for percutaneous vaccination, Vaccine,
2012https://doi.org/10.1016/j.vaccine.2011.11.111.
[147] K. Fukushima, A. Ise, H. Morita, R. Hasegawa, Y. Ito, N. Sugioka, K. Takada, Two-
layered dissolving microneedles for percutaneous delivery of peptide/protein
drugs in rats, Pharm. Res. (2011)https://doi.org/10.1007/s11095-010-0097-7.
[148] Z. Gui, X. Wu, S. Wang, Y. Cao, J. Wan, Q. Shan, Z. Yang, J. Zhang, S. Gui, Dissolving
microneedles integrated with liquid crystals facilitate transdermal delivery of
sinomenine hydrochloride, J. Pharm. Sci. (2017)https://doi.org/10.1016/j.xphs.
2017.07.027.
[149] Y. Ito, M. Taniguchi, A. Hayashi, M. Anai, S. Morita, E. Ko, N. Yoshimoto, Y. Yoshii, S.
Kobuchi, T. Sakaeda, K. Takada, Application of dissolving microneedles to glucose
monitoring through dermal interstitial fluid, Biol. Pharm. Bull. (2014)https://doi.
org/10.1248/bpb.b14-00406.
[150] H. Kim, K.Y. Seong, J.H. Lee, W. Park, S.Y. Yang, S.K. Hahn, Biodegradable
microneedle patch delivering antigenic peptide-hyaluronate conjugate for cancer
immunotherapy, ACS Biomater. Sci, Eng, 2019https://doi.org/10.1021/
acsbiomaterials.9b00961.
[151] Y. Qiu, C. Li, S. Zhang, G. Yang, M. He, Y. Gao, Systemic delivery of artemether by
dissolving microneedles, Int. J. Pharm. (2016)https://doi.org/10.1016/j.ijpharm.
2016.05.006.
[152] Z. Chen, B. Han, L. Liao, X. Hu, Q. Hu, Y. Gao, Y. Qiu, Enhanced transdermal delivery
of polydatin via a combination of inclusion complexes and dissolving microneedles
for treatment of acute gout arthritis, J. Drug Deliv. Sci. Technol. (2020)https://doi.
org/10.1016/j.jddst.2019.101487.
[153] J. Zhu, L. Dong, H. Du, J. Mao, Y. Xie, H. Wang, J. Lan, Y. Lou, Y. Fu, J. Wen, B. Jiang, Y.
Li, J. Zhu, J. Tao, 5-Aminolevulinic acid-loaded hyaluronic acid dissolving
microneedles for effective photodynamic therapy of superficial tumors with en-
hanced long-term stability, Adv. Healthc, Mater, 2019https://doi.org/10.1002/
adhm.201900896.
[154] H. Du, P. Liu, J. Zhu, J. Lan, Y. Li, L. Zhang, J. Zhu, J. Tao, Hyaluronic acid-based dis-
solving microneedle patch loaded with methotrexate for improved treatment of
psoriasis, ACS Appl. Mater. Interfaces (2019)https://doi.org/10.1021/acsami.
9b15668.
[155] G. Li, A. Badkar, S. Nema, C.S. Kolli, A.K. Banga, In vitro transdermal delivery of ther-
apeutic antibodies using maltose microneedles, Int. J. Pharm. (2009)https://doi.
org/10.1016/j.ijpharm.2008.10.008.
[156] C. Dillon, H. Hughes, N.J. O’Reilly, C.J. Allender, D.A. Barrow, P. McLoughlin, Dissolv-
ing microneedle based transdermal delivery of therapeutic peptide analogues, Int.
J. Pharm. (2019)https://doi.org/10.1016/j.ijpharm.2019.04.075.
[157] A. Donadei, H. Kraan, O. Ophorst, O. Flynn, C. O’Mahony, P.C. Soema, A.C. Moore,
Skin delivery of trivalent sabin inactivated poliovirus vaccine using dissolvable
microneedle patches induces neutralizing antibodies, J. Control. Release (2019)
https://doi.org/10.1016/j.jconrel.2019.08.039.
[158] S. Bhatnagar, S.R. Chawla, O.P. Kulkarni, V.V.K. Venuganti, Zein Microneedles for
Transcutaneous Vaccine Delivery: Fabrication, Characterization, and In Vivo Evalu-
ation Using Ovalbumin as the Model Antigen, ACS Omega, 2017https://doi.org/10.
1021/acsomega.7b00343.
[159] A. Aditya, B. Kim, R.D. Koyani, B. Oropeza, M. Furth, J. Kim, N.P. Kim, Kinetics of col-
lagen microneedle drug delivery system, J. Drug Deliv. Sci. Technol. (2019)https://
doi.org/10.1016/j.jddst.2019.03.007.
[160] W. Yu, G. Jiang, D. Liu, L. Li, Z. Tong, J. Yao, X. Kong, Transdermal delivery of insulin
with bioceramic composite microneedles fabricated by gelatin and hydroxyapatite,
Mater. Sci. Eng. C. (2017)https://doi.org/10.1016/j.msec.2016.12.111.
620
International Journal of Biological Macromolecules 170 (2021) 602–621
[161] W. Yu, G. Jiang, D. Liu, L. Li, H. Chen, Y. Liu, Q. Huang, Z. Tong, J. Yao, X. Kong, Fab-
rication of biodegradable composite microneedles based on calcium sulfate and
gelatin for transdermal delivery of insulin, Mater. Sci. Eng. C. (2017)https://doi.
org/10.1016/j.msec.2016.10.063.
[162] O. Olatunji, C.C. Igwe, A.S. Ahmed, D.O.A. Alhassan, G.O. Asieba, B. Das Diganta,
Microneedles from fish scale biopolymer, J. Appl. Polym. Sci. 131 (2014) 1–10,
https://doi.org/10.1002/app.40377.
[163] O. Olatunji, A. Denloye, Production of hydrogel microneedles from fish scale bio-
polymer, J. Polym. Environ. (2019)https://doi.org/10.1007/s10924-019-01426-x.
[164] O. Olatunji, R.T. Olsson, Microneedles from fishscale-nanocellulose blends using
low temperature mechanical press method, Pharmaceutics, 2015https://doi.org/
10.3390/pharmaceutics7040363.
[165] X. You, J.H. Chang, B.K. Ju, J.J. Pak, Rapidly dissolving fibroin microneedles for trans-
dermal drug delivery, Mater. Sci. Eng. C. (2011)https://doi.org/10.1016/j.msec.
2011.06.010.
[166] S. Wang, M. Zhu, L. Zhao, D. Kuang, S.C. Kundu, S. Lu, Insulin-loaded silk fibroin
microneedles as sustained release system, ACS Biomater. Sci, Eng, 2019https://
doi.org/10.1021/acsbiomaterials.9b00229.
[167] Z. Yin, D. Kuang, S. Wang, Z. Zheng, V.K. Yadavalli, S. Lu, Swellable silk fibroin
microneedles for transdermal drug delivery, Int. J. Biol. Macromol. (2018)https://
doi.org/10.1016/j.ijbiomac.2017.07.178.
[168] J.Y. Lee, S.H. Park, I.H. Seo, K.J. Lee, W.H. Ryu, Rapid and repeatable fabrication of
high A/R silk fibroin microneedles using thermally-drawn micromolds, Eur. J.
Pharm. Biopharm. (2015)https://doi.org/10.1016/j.ejpb.2015.04.024.
[169] N. Elharfaoui, M. Djabourov, W. Babel, Molecular weight influence on gelatin gels:
structure, enthalpy and rheology, Macromol. Symp. 256 (2007) 149–157, https://
doi.org/10.1002/masy.200751017.
[170] Q. Xing, K. Yates, C. Vogt, Z. Qian, M.C. Frost, F. Zhao, Increasing mechanical
strength of gelatin hydrogels by divalent metal ion removal, Sci. Rep. 4 (2014)
1–10, https://doi.org/10.1038/srep04706.
[171] C. Vepari, D.L. Kaplan, Silk as a biomaterial, Prog. Polym. Sci. 32 (2007) 991–1007,
https://doi.org/10.1016/j.progpolymsci.2007.05.013.
[172] P. Song, W. Zhang, C. Xu, H. Guan, Y. Tu, G. Wu, Effects of silkworm variety on the
mechanical and structural properties of silk, J. Chinese Adv. Mater. Soc. 6 (2018)
827–837, https://doi.org/10.1080/22243682.2018.1556334.
[173] D.C. Aduba, H. Yang, Polysaccharide fabrication platforms and biocompatibility as-
sessment as candidate wound dressing materials, Bioengineering. 4 (2017). doi:
https://doi.org/10.3390/bioengineering4010001.
[174] G. Bonfante, H. Lee, L. Bao, J. Park, N. Takama, B. Kim, Comparison of polymers to
enhance mechanical properties of microneedles for bio-medical applications,
Micro Nano Syst. Lett. 8 (2020) 13, https://doi.org/10.1186/s40486-020-00113-0.
[175] Y. Zhang, G. Jiang, W. Yu, D. Liu, B. Xu, Microneedles fabricated from alginate and
maltose for transdermal delivery of insulin on diabetic rats, Mater. Sci. Eng. C. 85
(2018) 18–26, https://doi.org/10.1016/j.msec.2017.12.006.
[176] Z. Cheng, H. Lin, Z. Wang, X. Yang, M. Zhang, X. Liu, B. Wang, Z. Wu, D. Chen, Prep-
aration and characterization of dissolving hyaluronic acid composite microneedles
loaded micelles for delivery of curcumin, Drug Deliv. Transl. Res. 10 (2020)
1520–1530, https://doi.org/10.1007/s13346-020-00735-2.
[177] Y.H. Chiu, M.C. Chen, S.W. Wan, Sodium hyaluronate/chitosan composite
microneedles as a single-dose intradermal immunization system,
Biomacromolecules. 19 (2018) 2278–2285, https://doi.org/10.1021/acs.biomac.
8b00441.
[178] M.H. Ling, M.C. Chen, Dissolving polymer microneedle patches for rapid and effi-
cient transdermal delivery of insulin to diabetic rats, Acta Biomater. 9 (2013)
8952–8961, https://doi.org/10.1016/j.actbio.2013.06.029.
[179] A.N. Cadinoiu, D.M. Rata, L.I. Atanase, Biocompatible injectable polysaccharide ma-
terials for drug delivery, in: Polysacch, Elsevier, Carriers Drug Deliv, 2019 127–154,
https://doi.org/10.1016/B978-0-08-102553-6.00006-4.
[180] E. Papakonstantinou, M. Roth, G. Karakiulakis, Hyaluronic acid: a key molecule in
skin aging, Dermatoendocrinol. 4 (2012) 253, https://doi.org/10.4161/derm.
21923.
[181] G. Kogan, L. Šoltés, R. Stern, J. Schiller, R. Mendichi, Hyaluronic acid: its function
and degradation in in vivo systems, Stud. Nat. Prod. Chem. 34 (2008) 789–882,
https://doi.org/10.1016/S1572-5995(08)80035-X.
[182] G.G. D’Ayala, M. Malinconico, P. Laurienzo, Marine derived polysaccharides for bio-
medical applications: chemical modification approaches, Molecules. 13 (2008)
2069–2106, https://doi.org/10.3390/molecules13092069.
[183] T. Sato, T. Ishii, Y. Okahata, In vitro gene delivery mediated by chitosan. Effect of
pH, serum, and molecular mass of chitosan on the transfection efficiency, Biomate-
rials. 22 (2001) 2075–2080. doi:https://doi.org/10.1016/S0142-9612(00)00385-9.
[184] S. Bhatnagar, P.R. Gadeela, P. Thathireddy, V.V.K. Venuganti, Microneedle-based
drug delivery: materials of construction, J. Chem. Sci. 131 (2019) 1–28, https://
doi.org/10.1007/s12039-019-1666-x.
[185] Jawaid Marichelvam, Asim, corn and rice starch-based bio-plastics as alternative
packaging materials, Fibers. 7 (2019) 32, https://doi.org/10.3390/fib7040032.
[186] Xanthan Gums | Cargill Pharmaceutical | Cargill, (n.d.). https://www.cargill.com/
pharmaceutical/hydrocolloids-for-pharma/xanthan-gums-for-pharma (accessed
December 15, 2020).
[187] D.F.S. Petri, Xanthan gum: a versatile biopolymer for biomedical and technological
applications, J. Appl. Polym. Sci. 132 (2015). doi:https://doi.org/10.1002/app.
42035.
[188] M.-B. Coltelli, S. Danti, K. De Clerck, A. Lazzeri, P. Morganti, Pullulan for advanced
sustainable body- and skin-contact applications, J. Funct. Biomater. 11 (2020) 20,
https://doi.org/10.3390/jfb11010020.
N. Dabholkar, S. Gorantla, T. Waghule et al.
[189] A.K. Lynn, I.V. Yannas, W. Bonfield, Antigenicity and immunogenicity of collagen, J.
Biomed. Mater. Res. - Part B Appl. Biomater. 71 (2004) 343–354, https://doi.org/10.
1002/jbm.b.30096.
[190] W. Yu, G. Jiang, D. Liu, L. Li, H. Chen, Y. Liu, Q. Huang, Z. Tong, J. Yao, X. Kong, Fab-
rication of biodegradable composite microneedles based on calcium sulfate and
gelatin for transdermal delivery of insulin, Mater. Sci. Eng. C. 71 (2017) 725–734,
https://doi.org/10.1016/j.msec.2016.10.063.
[191] W. Yu, G. Jiang, D. Liu, L. Li, Z. Tong, J. Yao, X. Kong, Transdermal delivery of insulin
with bioceramic composite microneedles fabricated by gelatin and hydroxyapatite,
Mater. Sci. Eng. C. 73 (2017) 425–428, https://doi.org/10.1016/j.msec.2016.12.111.
[192] A.A. El-Rashidy, G. Waly, A. Gad, J.A. Roether, J. Hum, Y. Yang, R. Detsch, A.A.
Hashem, I. Sami, W.H. Goldmann, A.R. Boccaccini, Antibacterial activity and bio-
compatibility of zein scaffolds containing silver-doped bioactive glass, Biomed.
Mater. 13 (2018). doi:https://doi.org/10.1088/1748-605X/aad8cf.
[193] J. Dong, Q. Sun, J.Y. Wang, Basic study of corn protein, zein, as a biomaterial in tis-
sue engineering, surface morphology and biocompatibility, Biomaterials. 25
(2004) 4691–4697, https://doi.org/10.1016/j.biomaterials.2003.10.084.
[194] T. Yucel, M.L. Lovett, D.L. Kaplan, Silk-based biomaterials for sustained drug deliv-
ery, J. Control. Release 190 (2014) 381–397, https://doi.org/10.1016/j.jconrel.2014.
05.059.
[195] R.F. Donnelly, of Microneedle-based Products, (2018) 307–322.
[196] Micro needle array-doxorubicin (MNA-D) in patients with cutaneous t-cell lym-
phoma (CTCL) - full text view - ClinicalTrials.gov, (n.d.). https://clinicaltrials.gov/
ct2/show/NCT02192021 (accessed July 7, 2020).
[197] Home - ClinicalTrials.gov, (n.d.). https://clinicaltrials.gov/ (accessed February 8,
2019).
[198] M. Kim, H. Yang, H. Kim, H. Jung, H. Jung, Novel cosmetic patches for wrinkle im-
provement: retinyl retinoate- and ascorbic acid-loaded dissolving microneedles,
Int. J. Cosmet. Sci. (2014)https://doi.org/10.1111/ics.12115.
[199] J.H. Lee, Y.S. Jung, G.M. Kim, J.M. Bae, A hyaluronic acid-based microneedle patch to
treat psoriatic plaques: a pilot open trial, Br. J. Dermatol. (2018)https://doi.org/10.
1111/bjd.15779.
[200] M. Avcil, G. Akman, J. Klokkers, D. Jeong, A. Çelik, Efficacy of bioactive peptides
loaded on hyaluronic acid microneedle patches: a monocentric clinical study, J.
Cosmet. Dermatol. (2020)https://doi.org/10.1111/jocd.13009.
[201] M. Jang, S. Baek, G. Kang, H. Yang, S. Kim, H. Jung, Dissolving microneedle with high
molecular weight hyaluronic acid to improve skin wrinkles, dermal density and
elasticity, Int. J. Cosmet. Sci. 42 (2020) 302–309, https://doi.org/10.1111/ics.12617.
621
International Journal of Biological Macromolecules 170 (2021) 602–621
[202] E. Peschl, K.W. Bauer, E. Peschl, K.W. Bauer, RU2701361C1, Über Nichtlineare Dif-
fer. 2. Ordnung, Die Für Eine Abschätzungsmethode Bei Partiellen Differ. Vom
Elliptischen Typus Bes. Wicht. Sind. 361 (1964) 18–30. doi:https://doi.org/10.
1007/978-3-322-98502-6_4.
[203] Japanese Inventor, JP2019523680A, JP2019523680A, 2017.
[204] R.F. Donnelly, D. Deliv, RU2016144039A, (2016) 1–21.
[205] T. Tepzz, H. Davon, EP2934660B1, 1 (2019) 1–28.
[206] E.M.P. David L. Kaplan, Konstantinos Tsioris, Fiorenzo G. Omenetto, CA2815285C,
CA2815285C, 2020.
[207] M. Bok, Z.J. Zhao, S. Jeon, J.H. Jeong, E. Lim, Ultrasonically and iontophoretically en-
hanced drug-delivery system based on dissolving microneedle patches, Sci. Rep.
(2020)https://doi.org/10.1038/s41598-020-58822-w.
[208] J. Madden, C. O’Mahony, M. Thompson, A. O’Riordan, P. Galvin, Biosensing in der-
mal interstitial fluid using microneedle based electrochemical devices, Sens, Bio-
Sensing Res, 2020https://doi.org/10.1016/j.sbsr.2020.100348.
[209] K.J. Lee, J.Y. Lee, S.G. Lee, S.H. Park, D.S. Yang, J.J. Lee, A. Khademhosseini, J.S. Kim,
W.H. Ryu, Microneedle drug eluting balloon for enhanced drug delivery to vascular
tissue, J. Control. Release (2020)https://doi.org/10.1016/j.jconrel.2020.02.012.
[210] O. Howells, N. Rajendran, S. Mcintyre, S. Amini-Asl, P. Henri, Y. Liu, O. Guy, A.E.G.
Cass, M.C. Morris, S. Sharma, Microneedle array-based platforms for future
theranostic applications, ChemBioChem, 2019https://doi.org/10.1002/cbic.
201900112.
[211] C.P.P. Pere, S.N. Economidou, G. Lall, C. Ziraud, J.S. Boateng, B.D. Alexander, D.A.
Lamprou, D. Douroumis, 3D printed microneedles for insulin skin delivery, Int. J.
Pharm. (2018)https://doi.org/10.1016/j.ijpharm.2018.03.031.
[212] S.N. Economidou, C.P.P. Pere, A. Reid, M.J. Uddin, J.F.C. Windmill, D.A. Lamprou, D.
Douroumis, 3D printed microneedle patches using stereolithography (SLA)for in-
tradermal insulin delivery, Mater. Sci. Eng. C. (2019)https://doi.org/10.1016/j.
msec.2019.04.063.
[213] K. Moussi, A. Bukhamsin, T. Hidalgo, J. Kosel, Biocompatible 3D printed
microneedles for transdermal, intradermal, and percutaneous applications, Adv.
Eng. Mater. (2020)https://doi.org/10.1002/adem.201901358.
[214] D. Han, R.S. Morde, S. Mariani, A.A. La Mattina, E. Vignali, C. Yang, G. Barillaro, H.
Lee, 4D printing of a bioinspired microneedle array with backward-facing barbs
for enhanced tissue adhesion, Adv. Funct. Mater. (2020)https://doi.org/10.1002/
adfm.201909197.