Materials Today Communications 32 (2022) 104109

Contents lists available at ScienceDirect

Materials Today Communications

journal homepage: www.elsevier.com/locate/mtcomm

A review of preparation methods of porous skin tissue engineering scaffolds

Zefei Zhang, Yihua Feng *, Li Wang, Dongxue Liu, Changcai Qin, Yanbin Shi
Qilu University of Technology (Shandong Academy of Sciences), School of Mechanical and Automotive Engineering, Jinan 250353, China

A R T I C L E I N F O A B S T R A C T

Keywords: In recent years, preparation technology for tissue-engineered scaffolds has continuously improved; researchers
Skin tissue engineering can select suitable preparation methods according to the scaffold requirements. A skin-tissue scaffold must have a
Scaffold porous structure; the structure affects wound healing, hydrophilicity, the degradation rate, and the mechanical
Porous structure
properties of the scaffold, and acts as a nutrient and metabolites transportation channel for cell adhesion and
Preparation method
growth, new tissue regeneration providing the space environment. Technologies including solvent casting/par­
ticle leaching, freeze-drying, electrospinning, 3-D bioprinting, and gas foaming have been used to prepare porous
skin scaffolds. Preparation using these five methods is examined in this review. The optimization design and
application of these methods are introduced; the advantages and disadvantages, and the future prospects of these
methods are summarized.

1. Introduction shapes by accurately controlling the printing path [9], as shown in

Skin is the first barrier of protection and the organ most vulnerable to
injury. The skin includes the epidermis, dermis, and subcutaneous tis­
sues[1], as shown in Fig. 1. Skin tissue engineering scaffolds are used to
guide skin repair when the skin is unable to repair itself. The main goal
of tissue engineering scaffolds is to repair and replicate damaged tissue
[2]. A suitable skin tissue-engineering scaffold must be able to support
the growth of new tissue and provide the host cells with a living space
suitable for cell migration and growth.
Studies have shown that an engineered scaffold with a structure
similar to that of the natural dermis is more suitable for wound healing
[3,4]. The overall structure and properties of porous materials largely
depend on the preparation method; pore size, porosity, and pore inter­
connection play an indispensable role in tissue regeneration. A
tissue-engineered scaffold should have a customized porous structure to
better adapt to cell proliferation and growth; the pore size affects the
growth of cells. The optimum pore size required for cell permeation
varies by cell type, the optimal pore size for adult mammal skin regen­
eration is 20–125 µm [5]. Larger pore sizes reduce intercellular in­
teractions and thus cell proliferation [6,7]. Inadequate pore size results
in poor cell permeability, impeding the transport of nutrients and
metabolic waste and preventing the formation of blood vessels [8]. To
evaluate the effect of pore size on the growth rate of cells, Xie et al.
prepared several heterogeneous scaffolds with different pore sizes and
Fig. 2. Cell alignment exhibited 4 different orientations in a single
scaffold with 4 regions, separately. The proliferation speed of the cells in
small pores is 3 times that of the cells in large pores. Some studies have
shown that scaffolds with 60–90% porosity are suitable for wound
healing applications as they are capable of providing sufficient space for
cell activity, oxygen and nutrient exchange, and the production of a new
extracellular matrix (ECM)[10]. Both low- and high-porosity scaffolds
support the early attachment, diffusion, and proliferation of primary
dermal fibroblasts, but only high-porosity scaffolds support the migra­
tion and infiltration of active cells [11]. An increase in the porosity of
the scaffold has a direct effect on the reduction in mechanical properties,
therefore the balance between the porosity of the scaffold and the me­
chanical properties is crucial. Khajeh et al. prepared skin scaffolds with
different porosity by varying the polymer concentration, the lower the
porosity, the better the mechanical properties, as shown in Fig. 3 AC,
and the cell viability under different pores is shown in Fig. 3B [12].
Meng et al. designed a scaffold with adjustable mechanical properties by
controlling the porous structure of the scaffold [13]. For this purpose,
the skin scaffold should have mechanical properties similar to those
found in native tissue. In this regard, the values of the tensile strengths,
the Young’s modulus, and the elongation-to-break are the most impor­
tant parameters for assessing the suitability of the mechanical properties
of the scaffold [14]. Although these values vary depending on the region
of the native tissue, according to the literature, the tensile strength

* Corresponding author.
E-mail addresses: 10431200071@stu.qlu.edu.cn (Z. Zhang), fyh@qlu.edu.cn (Y. Feng), wli@qlu.edu.cn (L. Wang), 10431200093@stu.qlu.edu.cn (D. Liu),
10431200044@stu.qlu.edu.cn (C. Qin), syb@qlu.edu.cn (Y. Shi).

https://doi.org/10.1016/j.mtcomm.2022.104109
Received 5 April 2022; Received in revised form 4 July 2022; Accepted 25 July 2022
Available online 26 July 2022
2352-4928/© 2022 Elsevier Ltd. All rights reserved.
Z. Zhang et al. Materials Today Communications 32 (2022) 104109

Fig. 1. Skin structure.
between 5 and 40 MPa, the Young’s modulus between 4.5 and 25 MPa,
and the elongation-to-break between 35% and 120% seem to be
appropriate for skin scaffold [15]. In practice, tissue-engineered
scaffolds with higher tensile strength and Young’s modulus will be more
stable in animal experiments. Researchers can change the porosity of the
scaffold to meet the mechanical property requirements [16]. Controlling
the porosity distribution over complex structures that produce func­
tional gradients, researchers can design optimal interconnect channels
based on functional gradients to enhance permeability and improve
fluid transport through the structure [17]. The size, geometry, direction,
interconnectivity, and surface chemistry of the channel determine the
nature of the nutrient flow. In addition, the interconnection of channels
assists in cell migration and tissue transport for endogenous growth.
Good connectivity allows fluid to be transported to deeper sides,
allowing biomaterials and trapped biomolecules to be released by
diffusion through these regions [18]. The porous structure of customized
scaffolds is important in building better human tissue engineering
scaffolds.
Wound healing has four overlapping stages: hemostasis, inflamma­
tion, proliferation and remodeling. The process of skin wound healing is
associated with a range of cellular responses, including coagulation,
platelet activation, macrophage infiltration, re-epithelialization and the
formation of granulation tissue (composed of fibroblasts and blood
vessels)[19]. Tissue engineering biomaterials often cause an

Fig. 2. Characteristics of scaffolds with different pore sizes. (A) Rectangular heterostructure with fiber spacing of 100 µm, 200 µm, and 500 µm, and crossover angle
of 90◦ ; (B) Heterostructure of parallelogram with fiber spacing of 100 µm and 200 µm, and crossover angle of 45◦ ; (C) Triangular heterostructure with fiber spacing of
100 µm and 200 µm, and intersection angle of 60◦ ; (D) (i) Diagram of cell growth induced by different pore sizes; (ii) Human umbilical vein endothelial cells in
scaffolds 100 µm and 200 µm in diameter on day 1; (iii) Human umbilical vein endothelial cells in scaffold with diameters of 100 µm and 200 µm on day 7; the pore
area is filled, and the macropore area remains empty. All stands are printed in five layers. (iv) Cell proliferation in different aperture regions, counting cells in four
600 µm x 600-micron regions
Reproduced with permission from [9].

2
Z. Zhang et al.
inflammatory response known as a foreign body response (FBR), medi­
ated by macrophages. Studies show that macrophages play a key role in
the success of tissue-engineered scaffold grafts[8]. Deonarine et al.
pointed out that macrophages in the early stages of skin wound healing
are of a mixed M1 and M2 phenotype while those that are M2 dominate
in the later stages[20]. This transformation not only avoids tissue
damage caused by chronic inflammation, but also promotes the recon­
struction of the surrounding tissue. Garg et al. obtained electrospinning
scaffolds with different pore sizes and showed that macrophage infil­
tration and the M2 phenotype can be significantly enhanced by culture
on a macroporous scaffold with a thick fiber diameter[21], suggesting
Fig. 4. Appearance (A) and SEM photomicrographs (B–K) of cross-sections of four types of collagen porous scaffolds prepared with ice particulates with diameter
ranges of 150–250 µm (B, G), 250–355 µm (C, H), 355–425 µm (D, I), and 425–500 µm (E, J), and a control collagen porous scaffold prepared without ice particulates
(F, K) at low (B, C, D, E, F) and high (G, H, I, J, K) magnifications.
Reproduced with permission from [33].
3
Materials Today Communications 32 (2022) 104109
Fig. 3. (A)The porosity of Polyurethane/Poly­
acrylic acid (PU/PAA), and PU/PAA nanofiber
containing different concentrations of Polox­
amer (POLO) (5, 10, and15 wt%). (B)Cell
viability of Polyurethane/Polyacrylic acid (PU/
PAA), and PU/PAA nanofiber containing
different concentrations of Poloxamer (POLO)
(5, 10,and 15 wt%). (C)The mechanical prop­
erties of Polyurethane/Polyacrylic acid (PU/
PAA), and PU/PAA nanofiber containing
different concentrations of Poloxamer (POLO)
(5, 10,and 15 wt%).
Reproduced with permission from [12].
that pore size plays an important role in tissue engineering scaffold
design. Fibroblasts are the main cellular component of the dermis and
the skin scaffold should promote adhesion, growth and migration of fi­
broblasts, thereby promoting regeneration of the dermis and epidermis.
Zhu et al. successfully prepared a microporous electrospinning scaffold.
In vitro experiments showed a significant increase in human dermal fi­
broblasts (HDF) on the macroporous scaffold, with HDFs growing into
the scaffold to a depth of more than 100 µm[22]. This microporous
scaffold promotes infiltration and proliferation of HDFs and is suitable
for skin tissue engineering. RnjakKovacina et al. obtained low and high
porosity synthetic human elastin scaffolds during electrospinning. The
Z. Zhang et al.
Fig. 5. Geometric accumulation of salt particles in polymer solutions of (A) conventional SCPL (no centrifugation) and (B) enhanced SCPL (centrifugation). Scanning
electron microscopy (SEM) showed the morphology of the polyurethane scaffold prepared using the enhanced SCPL method. Different solvent evaporation methods
and solvents were used. The salt grain size was 212–295 µm: (C) dimethylformamide/THF (50:50) air-drying; (D) dimethylformamide/THF (50:50) freeze-drying; (E)
dioxane–air-drying; (F) dioxane–freeze-drying
Reproduced with permission from [37].
scaffolds persisted for at least 6 weeks in a mouse subcutaneous im­
plantation study with fibroblasts on the exterior and infiltrating, evi­
dence of scaffold remodeling including de novo collagen synthesis and
early-stage angiogenesis[11]. For skin scaffolds with high porosity,
intra scaffold activity, collagen deposition, cell migration and infiltra­
tion within the scaffold can be achieved, leading to skin regeneration.
Therefore, designing a tissue-engineered scaffold that can use its porous
structure to modify the immune response and promote skin regeneration
is a promising strategy.
The effectiveness of tissue engineering scaffolds in tissue regenera­
tion depends mainly on the microstructure and morphology of the
porous structures (including porosity, pore size, pore shape, and in­
terconnections between pores) and the mechanical properties of the
scaffolds. After selection of appropriate materials, the first stage in tissue
engineering is the manufacture of porous three-dimensional scaffolds
[23]. Skin scaffolds include a porous membrane, a spongy body, and a
non-woven fiber net; scaffolds with different morphologies have
different microstructures. Skin tissue-engineered scaffolds can be pre­
pared using different methods depending on their geometry, material,
and microstructure requirements. Skin tissue engineering scaffolds with
porous structures have been constructed through solvent cas­
ting/particle leaching [24,25], freeze-drying [26], gas foaming [27],
electrostatic spinning[28], and 3-D bioprinting [29]. The following
sections describe how these five tissue-engineered scaffolds were pre­
pared and how the researchers optimized the scaffold’s porous structure.
2. Solvent casting/particle leaching
The technique of preparing porous scaffolds by dissolving particles of
different sizes and particle numbers is solvent casting/particle leaching.
Different materials have been used as pore-forming agents including
sodium chloride particles, sugar particles [6], ice particles [30], and
paraffin microspheres [31].
Traditional solvent casting/particle leaching is a popular research
method that controls the pore size and porosity of porous structures by
dissolving different sizes and numbers of particles [32]. Zhang et al.
accurately controlled the pore structure by changing the ice particle size
and freezing temperature, preparing four types of scaffolds with
different pore sizes [33], as shown in Fig. 4. In vitro experiments have
shown that scaffolds with 150–250 µm pore size are more densely
4
Materials Today Communications 32 (2022) 104109
packed with cells and more suitable for cell proliferation. Solvent cas­
ting/particle leaching is a simple process that does not require special­
ized equipment. Hu et al. used a particle leaching method with salt
particles and magnetic sugar particles as pore-forming agents to
manufacture sheet scaffolds with a controllable size range; they reported
that the scaffolds had potential for pore-forming agent arrangement,
pattern, and self-assembly [6]. High-porosity materials have difficulty
maintaining good mechanical properties; researchers have improved
membrane structure quality using different solvent systems or by adding
useful compounds. For example, deacetylchitosaccharides dissolved in
lactic acid are softer and more transparent than membranes dissolved in
glacial acetic acid [34]. Addition of plasticizers such as glycerin, poly­
ethylene glycol, and sorbitol further improve the tensile strength and
flexibility of the membrane, making it suitable for skin tissue engi­
neering applications[35].
The thickness limits the application of porous scaffolds. Mikos et al.
showed that solvent casting/particle leaching could only prepare ma­
terials with a thickness of 3 mm [36]. Large scaffolds require multilayer
superposition, which is difficult to prepare directly. Particle leaching is
often difficult with thick or large volume materials. Considering the
disconnection of internal pores and the presence of residual pores pro­
ducing particles inside the scaffold, Sin et al. developed an enhanced
solvent casting/particle leaching method for preparing
three-dimensional porous polyurethane (PU) scaffolds for cardiac tissue
engineering. The enhanced method combined the conventional solvent
casting/particle leaching method with centrifugal procedures [37], as
shown in Fig. 5. Centrifugation was used to improve the pore uniformity
and pore interconnection of the scaffolds. Porous structures in skin
tissue-engineered scaffolds can also be improved in this way. Chen et al.
used PLA/polyvinyl pyrrolidone (PVP) as raw materials for melt
blending, changing the blend ratio to improve the pore connectivity.
PVP is water-soluble with little organic solvent residue; a small amount
of residual polymer in ethylene carbonate annealing treatment is not
harmful to the body and helps to coarsen the behavior and increase the
aperture [38]. Flaibani combined a solvent casting/salting method with
anti-solvent precipitation. Micro- and nanoscale pores were synthesized
as polymer scaffolds, contributing to transport between cells and
allowing control of the macropore size by changing the sodium chloride
crystal size, and allowing gas processing by changing the polymer pre­
cipitation around the grain to improve the porosity [39]. Results from
Z. Zhang et al.
Fig. 6. (A) Percentage wound photographs and (B) wound size for control group, 2D mats and TNSs at various day intervals 0, 3, 5, 7, 10, 14 days. Data were
expressed as a percentage of the control value (mean ± SD).
Reproduced with permission from [45].
tests with mouse muscle cells show that the micro-nano porous scaffold
promotes cell adhesion to the polymer wall and enhances cell migration
and growth inside the scaffold.
Solvent casting/particle leaching technology controls the pore size
and porosity of the structure by dissolving different sizes and numbers of
particles. However, it is difficult to select a particle size for high-porosity
materials, and it is difficult to maintain good mechanical properties.
Moreover, the scaffold thickness is limited, the pores in the scaffold are
not connected, and there are residual particles in the scaffold that pro­
duce pores that must be addressed by other means. For example, adding
centrifugal steps can improve the uniformity and interconnection of the
bracket, and annealing is helpful to increase the pore size.
3. Freeze-drying
Freeze-drying method can avoid the disadvantages of limited thick­
ness and unconnected pores of solvent casting/particle leaching. The
freeze-drying method is commonly used in the preparation of scaffolds
and porous membranes in tissue engineering. Porous membranes pre­
pared using this method include collagen sponges, chitosan porous
scaffolds, and collagen–chitosan scaffolds, with high porosity and
adjustable pore size. Catanzano et al. produced highly porous inter­
connected alginate sponges with an average pore size of 300 µm through
freeze-drying [40].
The pore structure of freeze-dried scaffolds can be designed and
customized by controlling the formation of ice crystals during cooling.
The size, distribution, shape, and density of ice crystals can be controlled
by changing the freezing parameters, which means that the pore size,
gradient, shape, and porosity of the freeze-dried scaffolds can be
customized [41]. Pawelec et al. set different solution heights and limited
the temperature gradient to the axis of the solution to control the
5
Materials Today Communications 32 (2022) 104109
aperture and gradient distribution [42]. Frank et al. applied a magnetic
field to the solution system during the freezing process to induce the
growth direction of ice crystals to prepare directional porous scaffolds
[43]. Ghafari et al. prepared a double-layer skin scaffold with different
pores by controlling the polymer concentration to change the porosity
[44]. A lower polymer concentration results in higher porosity and
larger pore size of the underlayer, which is similar to the dermis of the
skin. A higher polymer concentration in the upper layer of the scaffold
results in lower porosity and smaller pore size, similar to the epidermal
layer of the skin. Yu et al. used dispersive freeze-drying to prepare
graded porous nano-scaffolds, obtaining large pores up to 40–150 µm in
diameter, and in vivo experiments showed that the skin scaffolds
significantly promoted fibroblast migration and infiltration and faster
wound recovery, as shown in Fig. 6 [45].
During the freezing process of water-soluble polymers, the size of ice
crystals is closely related to the freezing temperature and freezing rate;
thus, the freezing process determines the characteristics of the pores
after freeze-drying[46]. Lower temperature and faster freezing rate
result in smaller pore sizes of scaffolds[47,48]. By changing the blending
ratio and freezing temperature, Wang et al. showed a thin layer structure
with many slender pores inside the scaffold at − 20 ℃. At lower tem­
peratures, the scaffold exhibited a more regular and uniform pore
morphology, and more significant cell growth and proliferation[49].
In addition to adjusting the freezing rate and solution parameters,
crosslinking agents can also be used to enhance the material properties.
Fernandes-Silva prepared a sponge scaffold using marine collagen,
which was freeze-dried, modified with Genipin, and foamed with
hydrogel. The resulting scaffold had a highly porous and interconnected
structure [50]. Yang et al. successfully prepared a porous mixed scaffold
using a freeze-drying method with mixed porous silk fibroin protein, silk
fibroin protein, and hyaluronic acid, reporting that the prepared scaffold
Z. Zhang et al.
Fig. 7. (A) Design a of gel/Chs sponge scaffold and SEM images of porous sponge scaffolds with different solution ratios (B) formulation Chs; (C) formulation Gel/Chs
37; (D) formulation Gel/Chs 55; (E) formulation Gel/Chs 73; (F) formulation Gel.
Reproduced with permission from [54].
had enhanced mechanical properties, an appropriate swelling ratio and
supported cell proliferation and adhesion conducive to wound healing
[51]. Ran et al. synthesized a porous chitosan sponge and impregnated it
with porous aureus polysaccharides and silver nanoparticles. The in­
ternal pores were between 50 and 100 µm, with good expansion and
water retention to provide a moist environment for the wound. In the
wound healing test, the treatment group using sponge biomaterials
showed better wound contraction and tissue growth than the control
group, and good biocompatibility [52].
6
Materials Today Communications 32 (2022) 104109
Freeze-drying is also an important method for preparing porous
sponge scaffolds. Yuan et al. loaded lithium chloride (LiCl) into a chi­
tosan (CHI) hydrogel and prepared a sterile biocompatible sponge
scaffold by freeze-drying [53]. Fig. 7 shows that porous sponge scaffolds
prepared by freeze-drying with different solution ratios all have uniform
pore size, high porosity, high water absorption and retention capacity
and suitable degradation rates. The sponge scaffold can maintain its
pore structure during the whole process, providing effective cell support
and attachment for skin wound healing [54].
Fig. 8. (A) Average pore diameter and porosity
as a function of depressurization rate in super­
critical conditions of 40 ◦ C and 125 bar; (B)
Effect of pressure and temperature on pore size
with constant depressurization rate of 2.5 bar/s
and soaking time of 4 h; (C) Effect of pressure
and temperature on porosity with constant
depressurization rate of 2.5 bar/s and soaking
time of 4 h; (D) Microstructure of cross-
sectioned PPC–starch–bioglass disk foamed at
30 ◦ C and 75 bar depressurized at 2.5 bar/s
after 4 h of soaking.
Reproduced with permission from [62].
Z. Zhang et al.
Fig. 9. Diagram and experimental configuration of electrospinning. (A) Comparison of polymer melt and polymer solution processes; (B) MEW device: (1) pneumatic
assisted feeding system; (2) electric heating system; (3) high-voltage power supply; (4) computer-aided removable collecting plate; (5) syringe with molten polymer
and tip with electrode; (C) Address code motion path diagram and deposition process of fiber structure; (D) Fibrous submicron fibers deposited alternately in 0◦ and
90◦ directions.
Reproduced with permission from [75].
Freeze-drying preparation of the scaffolding allows customization of
the aperture, gradient, shape, and porosity; The porous structure of the
scaffold can be adjusted by changing the freeze-drying temperature,
freezing speed, and polymer concentration. By designing the tempera­
ture gradient of the cooling model, the vertical gradient distribution of
the pores can be made. The growth direction of ice crystals can be
induced by applying a magnetic field. The porous membrane porosity is
high, the pore size can be adjusted, and highly porous and inter­
connected scaffolds are possible. The disadvantages of limited thickness
and unconnected pores of solvent casting/particle leaching are avoided.
However, the pore structure is irregular; the mold design can be used to
control the heat transfer in the freeze-drying process to control the
framework.
4. Gas foaming
Gas foaming is a green, efficient, and environmentally friendly
method for preparation of porous materials; CO2 and N2 are inexpensive
and readily available, with no residue after foaming[55]. This is a
popular method for producing 3-D porous scaffolds[56–58]. However,
special defects are produced in CO2 foaming. Polymer materials such as
semi-crystalline or amorphous polymers always show a dense
non-foaming layer structure after preparation with microcellular
foaming technology [59,60]; it is difficult to meet the high porosity
requirements of scaffolds.
Pressure, temperature, decompression rate, and immersion time can
be changed to control the aperture and distribution of the gas foam to
optimize the 3-D scaffold. Horta et al. controlled the internal porosity by
regulating the pressure and decompression time to form interconnected
pores[61]. Manavitehrani et al. studied the application of a propylene
7
Materials Today Communications 32 (2022) 104109
carbonate starch-based scaffold in tissue engineering, optimizing the gas
foaming technique by adjusting parameters including pressure,
decompression rate, and temperature, effectively producing a porous
structure with high pore interconnection and cell and tissue infiltration
within the porous structure[62], as shown in Fig. 8. Ren et al. used
physical foaming technology with carbon dioxide as the foaming agent
and adjusted the initial film thickness, saturation pressure, and foaming
temperature to adjust the cellular morphology of polylactic acid porous
membranes. In mild conditions, polylactic acid porous films with highly
ordered straight channels and a porosity of approximately 72% were
obtained[63]. Chen et al. used gas foaming technology to improve the
porosity and loosely stacked fibrous structure of an electrowoven scaf­
fold with a larger pore size and a higher porosity and improved the
deposition of extracellular matrix components and the regeneration of
new blood vessels[64]. Zhang et al. developed a 3-D lamellar nanofiber
sponge. In the gas foaming process, the thickness and porosity of the
lamellar nanofiber structure can be finely controlled[58]. Hajiabbas
et al. prepared a double-layer nanocomposite scaffold with reasonable
porosity and pore size by combining electrospinning, in situ gas foam­
ing, in situ crosslinking, and freeze-drying[65]. Jing et al. used CO2
foaming technology combined with wet electrospinning to produce
three-dimensional electrospinning scaffold. The extracellular matrix
was completely simulated, in the porosity and surface area, and in the
surface nanomorphology. 3T3 fibroblast cell culture results confirmed
that the cells interacted strongly with the 3D fluffy nanofibers and
migrated into the inner area of the scaffolds rapidly. Moreover, human
fibroblast cell culture results confirmed further enhancement in cell
attachment and proliferation on the fluffy shish-kebab structured
nanofibrous scaffolds[66], which indicates scaffold with similar tissue
structure better promotes 3D tissue regeneration.
Z. Zhang et al. Materials Today Communications 32 (2022) 104109

Fig. 10. Schematic illustrates the design of the different pore structured PCL scaffolds: homogeneous pore size constructs of 250, 500, 750 µm;offset 30/70; offset
50/50; gradient pore structure. and SEM images of scaffolds with different porous structures (A) uniform pore sizes of 250 µm, (B)500 µm, and (C)750 µm; (D)Offset
30/70 with 150/30-micron pores; (E)Offset 50/50 with 250/250-micron pores; (F) Gradient pore structure (bottom: 250 µm, middle: 500 µm, top: 750 µm).
Reproduced with permission from [80].

8
Z. Zhang et al. Materials Today Communications 32 (2022) 104109

Fig. 11. Confocal microscope images and cross-section analysis of cells cultured (deposited) for 1 day on mesh sizes of (A, B) 100 µm; (C, D) 200 µm; (E, F) 300 µm
Reproduced with permission from [81].

9
Z. Zhang et al.
Gas foaming is a green, efficient, and environmentally friendly
porous material preparation method, which does not produce residual
pore-causing particles like solvent casting/particle leaching. Gas foam­
ing method can adjust the porous structure by adjusting the pressure,
decompression rate, temperature, and other parameters. But produces
special defects in the foaming of polymer samples, which makes it
difficult to achieve high porosity. However, the combination of wet
electrostatic spinning can achieve the desired pore structure. Therefore,
This technology should be used in combination with other methods or as
an auxiliary method.
5. Electrospinning
Electrospinning is a widely used technique in many research fields,
especially in the biomedical field[67]; it can produce fibers from milli­
meters to nanometers in diameter. Electrospun nanofibers have a high
surface area to volume ratio and a swelling property[68], good for
wound healing.
Electrospinning uses a strong electric field to process polymer solu­
tions into thin and continuous polymer fibers similar in shape to natural
extracellular matrix fibers to promote cell interactions and facilitate new
tissue formation[69]. In solution electrospinning, nanofiber structures
are randomly arranged on the receiver. The electrospinning results are
closely related to a number of factors including polymer solution
properties (viscosity/concentration, conductivity, surface tension, and
solvent), processing variables (electric field strength, solution feed rate,
distance between the spinning pore and the collecting device), and
environmental conditions (temperature and humidity)[70]. Zhang et al.
found that there was a direct relationship between the fiber morphology
and the concentration change, and prepared a superfine gelatin (Gt)
fiber[71]. The porous structure of the scaffold was a key factor affecting
cell attachment. The electrospinning mesh showed excellent in vitro
compatibility, but its relatively small pore size did not allow cell
migration in the scaffold (the cell diameter (5–20 µm))[72]; other
methods were required to open the pore. Kim et al. prepared bimodal
scaffolds with small and large diameter elements through solution
electrospinning and melt electrospinning. The larger pore size was
randomly mixed in the composite structure, allowing free migration of
cells [72]. Huang et al. prepared skin scaffolds with a gradient pore size
by combining multi-step electrospinning with a low-temperature
collection. They adjusted the gradient pore size by controlling the fila­
ment concentration, electric field, flow rate, needle pitch, and collector
temperature, and prepared skin scaffolds with a gradient pore structure
composed of small, medium, and large pores[73]. The optimum relative
humidity of the large pore size fraction is 50%, which significantly im­
proves the infiltration of cells into the scaffold. The medium pore size
fraction is more favorable to cell proliferation than the large pore frac­
tion. The small pore size fraction has higher mechanical properties and
inhibits surface cells.
Replacing a polymer solution with a highly viscous and non-
conductive polymer melt is a means of greatly reducing the surface
charge density to suppress any bending instability experienced by the
melt jet [74] (Fig. 11A). The application range is narrower than solution
electrospinning; the requirements for spinning devices are greater and
the prepared fibers are coarse. Melt electrospinning is mainly used to
meet medical requirements and standards such as the absence of solvent
toxicity.
Near-field direct writing technology addresses some of the limita­
tions of electrospinning[75–77]. Conventional electrospinning can only
produce a disordered fiber mass, and cannot obtain the scaffold of a
specific structure, as shown in Fig. 9. When the nozzle is close to the
receiving device, the Taylor cone is suppressed, and a straight fiber can
be obtained. With the controllable motion of the three-dimensional
moving platform, an orderly 3-D structure can be obtained [78]. Scaf­
fold products produced using this technique can have larger pore sizes
and are more suitable for cell invasion and growth. Farrugia et al.
10
Materials Today Communications 32 (2022) 104109
combined melt electrospinning with a programmable X–Y platform, The
scaffolds obtained have a highly porous three-dimensional structure
[72]. The features of the scaffolds including shape, pore size, porosity,
and mechanical properties can be controlled by the input parameters
[79]. Hochleitner et al. used the melt electrospinning writing (MEW)
device (Fig. 9B) to optimize the flow rate, spinneret diameter, voltage,
collector distance, and other parameters, and designed the print path
(Fig. 9C) to directly prepare the coherent scaffold of the ultrafine [75].
Abbasi et al. constructed scaffolds with different porous structures,
as shown in Fig. 10, and examined the effects of pore size, scaffold
migration, and gradient structure on the attachment, migration, and
proliferation of human osteoblasts. Among these scaffolds, the fibrous
offset scaffolds promoted osteoblast separation as a result of their large
surface area to volume ratio. Scaffolds with uniform 250-micron pores
showed the greatest attachment of osteoblasts, confirming the effect of
porous tissue on the behavior of cells in the scaffold[80]. These effects
persist on skin cells.
However, the fiber scaffold prepared by fused electrospinning
exhibited obvious drooping behavior. Nguyen et al. proposed an
experimental method known as the "beam bridge test" to determine the
drooping behavior of fused electrospinning microfibers. In fused elec­
trospinning conditions, the maximum pitch size corresponding to the
mesh size without drooping in the mesh pattern can be easily identified
using the beam bridge test[81]. Droop information can be used to
fabricate the lattice structure of microfibers with well-defined transverse
pores, and the prepared microfiber mesh structure can be successfully
used as a matrix (or scaffold) for cell culture. Drooping of microfibers
and the mesh size of the grid pattern significantly affected cell growth,
as shown in Fig. 11; the crisscross area indicates excellent cell attach­
ment. These results showed that microfibers without droop exhibited
excellent cell attachment.
The precision with which a fused electrospun scaffold can be man­
ufactured is greatly influenced by the deposited pattern (including the
spacing between the fibers), the velocity of translation, and the accu­
mulation of residual charges in the fibers. Greater understanding of how
the viscous charged melt electrospinning jet interacts with the collector
surface improves the accuracy and repeatability of fiber deposition [82].
Molten electrospinning has the potential to greatly advance the field of
tissue engineering. However, the complexity of the machine/software
and the amount of time required to manufacture a melt-write scaffold
prevent widespread use[83].
Electrostatic spinning technology prepares fibers from millimeters to
nanometers in diameter, changing the polymer concentration, voltage
intensity, solution flow rate and so on can control the fiber diameter;
however, the pore size is too small and the pore structure is random.
With melt electrostatic spinning, there is no solution solvent toxicity or
toxic accumulation, but the scope of application is narrow, there are
high requirements for the spinning device, and the fiber is thick. Melt
direct writing technology can be used to obtain scaffolds with specific
structures, allowing for a larger pore size more suitable for cell invasion
and growth. Further research should improve the resolution of melt
electrospinning to replicate the "fine" morphology of natural tissues.
6. 3-D bioprinting
3-D printing technology has high intelligence, fast processing spe­
Compared with traditional material preparation methods, ed, fine
structure, and good controllability[84–86]. 3-D printing of porous
structures requires a CAD model to be designed in advance; the shape
characteristics, size, and porosity can be designed according to the re­
quirements of the designing software. 3-D bioprinting is an important
branch of 3-D printing technology, but involves additional complexities
including bio-inks, cell types, selection of bioactive molecules, and
technical challenges associated with the sensitivity of living cells and
complex tissue and organ structures[87]. The main types of bio-printing
are inkjet, micro-extrusion, and laser-assisted bio-printing[88], as
Z. Zhang et al.
Fig. 12. 3-D bio-printing technologies. (A)Laser auxiliary printer; (B)Inkjet bioprinter; (C)Micro-extrusion printer. In all three printers, precise control of the X-, Y-,
and Z-coordinates is maintained to create the required 3-D biological structures
Reproduced with permission from [89].
Fig. 13. 3-D bioprinting operation.
Reproduced with permission from [92].
shown in Fig. 12. 3-D bio-printing is a new technology that delivers cells
from polymer gels to full-thickness skin wounds.
The printing parameters must be strictly controlled during the
printing process to obtain the best product. 3-D bioprinting parameters
include extrusion pressure, nozzle diameter, dispensing speed, bio­
printing speed, and bio-fabrication time. Bio-fabrication time de­
termines the embedding of cells in biological printing according to
exposure time. The print speed strongly influences the structure of cell
viability. The biological ink viscosity, the nozzle diameter and faster
printing speed combination that can be made in a shorter period of time
for load cell scaffolds by high resolution 3-D printing[90]. Bio­
printingoffers the ability to deliver cells in a fast, off-the-shelf manner,
helping to quickly cover and close wounds [91]. The printing process is
illustrated in Fig. 13.
Bioprinting of skin tissue depends on the bio-ink design, which
should support cellular compatibility and cellular function, and promote
production of new extracellular matrix components. The structure of the
skin tissue depends on the sequence of fibroblasts and keratinocytes
embedded in the bio-ink in the layered structure. Bio-ink is a solution
containing nutrients, matrix components, and cells. When developing
bio-inks for 3-D bioprinting, a number of factors should be considered to
balance printability and biocompatibility. As with biomaterial inks, the
11
Materials Today Communications 32 (2022) 104109
printing suitability of bio-inks is determined by the rheological prop­
erties, cross-linking mechanism, and 3-D printing parameters[93]. The
deposition of bio-inks is controlled with micron-scale precision to form
different tissue ultrastructures; 3-D bioprinting allows for an ideal
spatial distribution of cell-loaded scaffolds[94].
Scaffolds prepared with 3-D bioprinting are similar in complexity
and heterogeneity to those found in natural tissues, and can be arranged
into 3-D structures with cells and soluble growth factors in bio-inks,
providing new opportunities for implant designs with tissue regenera­
tion functions[92]. Cubo et al. used extruded bioprinting to print
bio-inks containing human plasma, human fibroblasts, and keratino­
cytes. In vitro and in vivo studies have shown that 3-D-printed skin
tissue is similar to human skin tissue[95]. 3-D bioprinting can accurately
print matrix materials and a variety of skin cells to the design position
through accurate positioning and transportation of living matter units
such as cells and extracellular matrix materials. The pipe network
structure can realize the self-supply of nutrients and waste metabolism
and transportation of skin[95]. Biological 3-D printing technology pro­
vides new options for precise bionic repair of skin defects[96,97]. Skin is
a tissue composed of two layers, the dermis and epidermis. The dermis is
a thicker, lower layer with higher flexibility and porosity. The epidermis
is a thin upper layer with low porosity, forming a protective barrier
Z. Zhang et al. Materials Today Communications 32 (2022) 104109

Fig. 14. (A, B) CAD images of native skin structure: the upper layer is dense and resembles the epidermis, and the lower porous layer resembles the dermis. (C) DLP-
based 3-D printing product showing the complex structure of upper and lower layers. The side view shows two layers of organization
Reproduced with permission from [98].

Fig. 15. (A) Skin defects divided into four groups: control group (untreated), hydrogel group, scaffold group, and FLS group; (B) Macro assessment was performed at
0 d, 4 d, 7 d, 14 d, and 21 d postoperatively; (C) Percentage of wound closure on day 4, 7, 14, and 21 post-operation. [98]
Reproduced with permission from [98].

against pathogenic microorganisms in the environment[44]. Functional
living skin (FLS) designed by Zhou et al. was composed of two layers
(Fig. 14): a dense upper layer mimicking the epidermal layer and a
porous lower layer mimicking the dermis, according to the physiological
structure of natural skin. A new method was developed to print FLS; it
was demonstrated that FLS has interconnected microchannels and
excellent performance in promoting dermal regeneration[98]. In addi­
tion, a highly porous and thick layer of scaffolding that mimics the
dermis absorbs wound exudates, reducing the risk of infection. The FLS
promotes skin regeneration by mimicking the physiological structure of
natural skin(Fig. 15). Niu et al. constructed a porous gradient whole skin
scaffold using bioprinting technology and tested the results using a
porcine model, with controlled results demonstrating that the scaffold
group repaired without fibrosis and with complete regeneration of the
epithelial layer[99].
There is growing research on the combination of 3-D printing with
other manufacturing methods. Researchers have recently combined 3-D
printing with electrospinning to produce tissue-engineered scaffolds.
Mori et al. produced composite hydrogels for tissue engineering with 3-
D printing and electrospinning[100]. The 3-D structure simulates the
structure and properties of the natural skin extracellular matrix in detail,
increasing cell proliferation.
3-D printing, relative to all other techniques, has the obvious
advantage that products can be designed with precise detail in terms of
the pore shape, arrangement, size, and structure, as the final pore
structure is not dependent on an emulsion, fluid mechanics, thermody­
namics (the matrix pore forming agent distribution), nor is it a random
event (gas foaming and electrostatic spinning)[101]. Studies have
shown that the 3-D printing process has few adverse effects on cell
survival[102]. CAD can only design regular microporous structures,
whereas human tissue is irregular, with different porous structures in
different locations[103]. Due to large solid particles tend to clog printer
nozzles, large aperture 3D bioprint brackets are challenging in practice
[104]. The field of 3-D printing is expanding; 3-D-printed scaffolds have

12
Z. Zhang et al. Materials Today Communications 32 (2022) 104109

Table 1
The influence of each parameter on pore structure.
The Parameter Influence Advantage Disadvantages Reference
preparation
method

Solvent Particle size Pore size is positively correlated The pore size and porosity of porous The internal pores of the scaffolds are [32,33,38,
casting/ with particle diameter structures are controlled by dissolving not connected and contain residual 108]
particle Number of The porosity is positively particles of different sizes and the pore-forming particles
leaching particles correlated with the number of number of particles
particles
Freeze-drying The freezing Pore size is positively correlated The pore size, gradient, shape, and The pore structure is irregular [44,46–48,
temperature with temperature, the lower the porosity in the scaffold can be adjusted 51,107]
temperature, the more uniform the as needed
pore
Freezing rate Pore diameter is negatively
correlated with freezing rate
Polymer Polymer concentration is
concentration negatively correlated with pore
size and porosity
Gas foaming Pressure Pore size is positively correlated The porous structure can be changed by There is a dense non-foaming cortex [55,59,60,
with pressure controlling the pressure and structure， pore formation is random 62]
Decompression Pore size and porosity are decompression rate, increasing the
rate negatively correlated with temperature to obtain a larger pore size.
decompression rate. CO2 has low cost, abundant source, and
temperature Pore size is positively correlated no residue after foaming
with temperature
Electrostatic polymer At too low a concentration， Electrospinning is a technique for The pore diameter of traditional [70–75,109]
spinning concentration microbeads or beaded nanofibers preparing millimeter-diameter to nano- electrospinning is too small and the
form. Too high a concentration scale fibers, preferably at a minimum porous structure is random, the
will block the flow of the solution flow rate to maintain a balance between application range of melt
through the tip the polymer solution and the new electrospinning is narrow, and the
Applied voltage Fiber diameter is negatively solution during jet formation. The spinning device is highly required.
correlated with voltage scaffolds prepared by melt
Solution flow rate Too low flow rate will lead to the electrospinning allow a larger aperture,
increase of pore size and fiber and the fibers prepared are thicker,
diameter, too high will lead to the which are more suitable for cell
formation of microbeads invasion and growth.
Needle to The diameter of nanofibers is
collector distance negatively correlated with
distance
Ambient relative The pore size increased with
humidity（RH） the improvement of RH within a
suitable range
3-D bioprinting Nozzle diameter The print resolution is negatively High degree of intelligence, fast CAD can only design regular micropore [90,91,93,
correlated with nozzle diameter processing speed, fine structure, good structure, can not ensure that the pore 94,96,97,
The speedy The speedy gelatinization is control, allowing the printing of structure and real skin are completely 101–103]
gelatinization conducive to the rapid molding of biological materials in an automated consistent
the desired 3D scaffolds with high and reproducible manner with high
resolution resolution and accuracy
Bioprinting speed Fiber diameter is negatively
correlated with velocity
Bio-fabrication The exposure time of embedded
time cells under bio-printing conditions
is determined
Solution Too low, the structure tends to
concentration collapse; Too high, the printing
process will produce sliding
behavior

been used in clinical studies with varying degrees of success[105,106]. 7. Discussion

3-D bioprinting technology offers high intelligence, fast processing
speed, fine structure, and good controllability, allowing the printing of
biological materials in an automatic and reproducible manner with high
resolution and accuracy, including complex 3-D structures with
biomechanical heterogeneity. However, the development of functional
3-D tissue constructs remains challenging, including the precise locali­
zation of different types of cells, growth factors, and biomaterials to
simulate the microenvironment in vivo. Moreover, CAD can only design
regular microporous structures, and the porosity cannot be guaranteed
to match the porosity of the cells. The next step of research should focus
on optimizing the printing path and improving its accuracy to prepare
scaffolds with complex textures.
The number of cases of tissue or organ dysfunction or loss caused by
trauma and chronic diseases is increasing worldwide, and tissue engi­
neering has unique advantages in repairing and treating such defects.
Skin scaffolds grafting is a new skin repair technology. In the aspect of
tissue engineering, tissue engineering scaffold is one of the focuses of
tissue engineering research. It provides space and effective physical
support for cell adhesion and growth and determines the structure and
shape of the target tissue or organ. At present, how to construct a three-
dimensional scaffold for promoting cell invasion and skin tissue repair is
still a hot issue in the field of tissue engineering.
Scaffolds must have sufficient porosity, average pore size, and
interconnectivity[107]. The porosity allows cells to penetrate the 3-D
structure and promote tissue regeneration. Porous interconnections

13
Z. Zhang et al.
Fig. 16. Summary of optimization design of various preparation methods.
ensure adequate diffusion of nutrients into the building body, and
diffuse metabolic wastes out of the scaffold. The manufacturing pa­
rameters are important in the preparation of tissue-engineered scaffolds.
The influence of each parameter on pore structure is shown in Table 1.
Porous skin scaffolds are in a rapid development stage. Many im­
provements have been made in the preparation of porous skin scaffolds,
encouraging the rapid development of tissue engineering and regener­
ative medicine, as shown in Fig. 16. Porous skin scafsfold are also facing
many challenges. First, the traditional manufacturing of functionally
graded porous scaffolds has many problems, such as difficult control­
lability of pores and mechanical properties, low precision, difficult
repetition, and so on. The mechanical properties of skin scaffolds are
usually much higher than those of real skin tissue, which may lead to
mechanical biological mismatch. Due to scaffold degradation and long-
term deformation in vivo, scaffolds may lose their mechanical properties
before tissue regeneration and maturation, affecting wound healing
outcomes. Therefore, future research should also focus on systematically
investigating the dynamic relationship between scaffold degradation
and mechanical properties to determine the optimal initial strength and
further design the optimal porous structure. Secondly, it is difficult to
compare those dressings from the aspect of wound recovery. The main
reason is that those dressings with different structures could not meet
the requirements of different wounds at the same time, which requires
researchers to investigate the wound healing effect of dressings more
specifically, instead of one wound model for all wounds[110]. Thirdly, it
is still a challenge to regenerate skin with normal appearance and
function[111]. Researchers should find a suitable preparation method to
adjust the porous structure of skin scaffolds to practical applications and
combine bioactive molecules and biomaterials that promote tissue
regeneration to produce skin scaffolds that better meet expectations.
To facilitate the translation of these novel porous skin scaffolds from
the laboratory to the clinic, their safety and efficacy need to be further
validated in a variety of large animal models (such as pig or monkey
14
Materials Today Communications 32 (2022) 104109
models) before they can enter clinical trials. At the same time, their
physicochemical and biological properties such as in vivo degradability,
immunogenicity and pro-angiogenic activity need to be critically eval­
uated to meet the requirements for clinical application. With the prog­
ress in the field of biomaterials and tissue engineering and
multidisciplinary collaborative efforts, the preparation of porous skin
scaffolds will continue to develop and innovate in the future. The con­
struction of products with more complex structures and bionic skin
functions will further facilitate disease treatment and tissue regenera­
tion, and then promote the clinical medical practice.
Cellular infiltration is the key to replacing scaffolds with new tissues,
and as human physiological microenvironment is regulated by many
factors, the traditional preparation methods are not reproduce the dy­
namic microenvironment of natural tissue. The author predicts that the
preparation methods based on electrospinning and 3D printing tech­
nology will be more popular in the future. The use of electrostatic
spinning and 3-D bioprinting technologies allows for the synthesis of
nanostructured scaffolds at both the molecular and atomic level, while
allowing for precise control of the scaffold’s porous structure, facili­
tating the field of tissue engineering. In addition, multi-material bio­
printing is another promising solution for recreating the complex
composition, structure, and function of natural tissues. Integrating 3D
printing technology with electrostatic spinning to prepare multi-
material and multi-scale structures in a hybrid manner is an effective
way to prepare porous skin scaffolds. At the same time, skin tissue en­
gineering scaffolds are modified using chemical cross-linking methods to
improve their properties in terms of morphology, mechanical strength,
surface wettability, compatibility, and biocompatibility.
8. Conclusion
Tissue engineering and regenerative medicine are emerging research
fields, including the use of different synthetic techniques and
Z. Zhang et al.
biocompatible components to design different types of porous skin
scaffolds for the migration and growth of skin cells. The effectiveness of
tissue-engineered scaffolds in assisting tissue regeneration depends
mainly on the microstructure and morphology of the porous structure
(including porosity, pore size, pore shape, the interconnection between
pores) and the mechanical properties of the scaffold. This paper focuses
on five preparation methods for skin tissue engineering scaffolds,
including solvent casting/particle leaching, freeze-drying, gas foaming,
electrostatic spinning, and 3-D bioprinting. The optimal design and
application of various techniques for scaffold porous structures are
presented, and the advantages and disadvantages of each technique and
future perspectives are summarised.
This review highlights the importance of porous structures for skin
scaffolds, where high porosity facilitates skin tissue cell adhesion/pro­
liferation/differentiation and low porosity ensures adequate mechanical
properties are required in the bearing zone, summarises the influence of
preparation parameters on scaffold pore structure and biological effects,
gives the authors’ expectations for the development of preparation
techniques, and the paper anticipates that hybrid preparation of multi-
materials based on electrostatic spinning and 3-D printing and multi­
scale structures based on electrostatic spinning and 3-D printing are
expected to be more interesting. For large-scale production, there is also
a need for biomaterials that are readily available, easy to manufacture,
and relatively inexpensive. Biomaterials with enhanced physiological
relevance are produced using biochemical mechanisms. Developments
in preparation technology have led to significant performance im­
provements in one of the three elements of tissue engineering,
’materials’.
In summary, research at the intersection of materials science,
structural design, fabrication, and tissue engineering has provided
methods for the preparation and use of skin tissue engineering scaffolds.
Substantial progress has been made in the fabrication of more complex
and accurate skin structures. However, the successful preparation of skin
scaffolds that achieve skin tissue-like appearance and function remains a
challenge. The development of innovative material designs and
advanced preparation processes to prepare skin scaffolds with structures
resembling real skin and excellent mechanical properties remains a
focus for future research.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This work is supported by Key Research and Development Plan of
Shandong Province [2018GGX103006], the National Natural Science
Foundation of China [51675289].
Author statement
No conflict of interest exits in the submission of this manuscript, and
the manuscript is approved by all authors for publication. I would like to
declare on beinal study that has not been published previously, and it is
not under consideration for publication elsewhere, in whole or in part.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data Availability
Data will be made available on request.
15
Materials Today Communications 32 (2022) 104109
References
[1] H.E. Abaci, Z. Guo, Y. Doucet, J. Jackow, A. Christiano, Next generation human
skin constructs as advanced tools for drug development, Exp. Biol. Med
(Maywood) 242 (2017) 1657–1668. 〈https://doi.10.1177/1535370217712690〉.
[2] Z. Pedram Rad, J. Mokhtari, M. Abbasi, Fabrication and characterization of PCL/
zein/gum arabic electrospun nanocomposite scaffold for skin tissue engineering,
Mater. Sci. Eng. C. Mater. Biol. Appl. 93 (2018) 356–366, https://doi.10.1016/j.
msec.2018.08.010.
[3] Y. Wang, R. Xu, G. Luo, Q. Lei, Q. Shu, Z. Yao, H. Li, J. Zhou, J. Tan, S. Yang,
R. Zhan, W. He, J. Wu, Biomimetic fibroblast-loaded artificial dermis with
“sandwich” structure and designed gradient pore sizes promotes wound healing
by favoring granulation tissue formation and wound re-epithelialization, Acta
Biomater. 30 (2016) 246–257, https://doi.10.1016/j.actbio.2015.11.035.
[4] S. Haldar, A. Sharma, S. Gupta, S. Chauhan, P. Roy, D. Lahiri, Bioengineered
smart trilayer skin tissue substitute for efficient deep wound healing, Mater. Sci.
Eng. C. Mater. Biol. Appl. 105 (2019), 110140 https://doi.10.1016/j.
msec.2019.110140.
[5] I.V. Yannas, E. Lee, D.P. Orgill, E.M. Skrabut, G.F. Murphy, Synthesis and
characterization of a model extracellular matrix that induces partial regeneration
of adult mammalian skin, Natl. Acad. Sci. 86 (1989) 933–937, https://
doi.10.1073/pnas.86.3.933.
[6] C. Hu, C. Tercero, S. Ikeda, M. Nakajima, H. Tajima, Y. Shen, T. Fukuda, F. Arai,
Biodegradable porous sheet-like scaffolds for soft-tissue engineering using a
combined particulate leaching of salt particles and magnetic sugar particles,
J. Biosci. Bioeng. 116 (2013) 126–131, https://doi.10.1016/j.
jbiosc.2013.01.011.
[7] J. An, J.E.M. Teoh, R. Suntornnond, C.K. Chua, Design and 3D printing of
scaffolds and tissues, Engineering 1 (2015) 261–268. 〈https://doi.10.15302/j-en
g-2015061〉.
[8] Y. Zhang, M. Zhang, D. Cheng, S. Xu, C. Du, L. Xie, W. Zhao, Applications of
electrospun scaffolds with enlarged pores in tissue engineering, Biomater. Sci. 10
(2022) 1423–1447. 〈https://doi.10.1039/d1bm01651b〉.
[9] C. Xie, Q. Gao, P. Wang, L. Shao, H. Yuan, J. Fu, W. Chen, Y. He, Structure-
induced cell growth by 3D printing of heterogeneous scaffolds with ultrafine
fibers, Mater. Des. 181 (2019), 108092 https://doi.10.1016/j.
matdes.2019.108092.
[10] T.M. Freyman, I.V. Yannas, L.J. Gibson, Cellular materials as porous scaffolds for
tissue engineering, Prog. Mater. Sci. 46 (2001) 273–282, https://doi.10.1016/
S0079-6425(00)00018-9.
[11] J. Rnjak-Kovacina, S.G. Wise, Z. Li, P.K. Maitz, C.J. Young, Y. Wang, A.S. Weiss,
Tailoring the porosity and pore size of electrospun synthetic human elastin
scaffolds for dermal tissue engineering, Biomaterials 32 (2011) 6729–6736,
https://doi.10.1016/j.biomaterials.2011.05.065.
[12] H. Gharib Khajeh, M. Sabzi, S. Ramezani, A.A. Jalili, M. Ghorbani, Fabrication of
a wound dressing mat based on Polyurethane/Polyacrylic acid containing
Poloxamer for skin tissue engineering, Colloids Surf. A: Physicochem. Eng. Asp.
633 (2022), 127891 https://doi.10.1016/j.colsurfa.2021.127891.
[13] Z. Meng, J. He, Z. Cai, F. Wang, J. Zhang, L. Wang, R. Ling, D. Li, Design and
additive manufacturing of flexible polycaprolactone scaffolds with highly-tunable
mechanical properties for soft tissue engineering, Mater. Des. 189 (2020), 108508
https://doi.10.1016/j.matdes.2020.108508.
[14] H.U. Zaman, J.M. Islam, M.A. Khan, R.A. Khan, Physico-mechanical properties of
wound dressing material and its biomedical application, J. Mech. Behav. Biomed.
Mater. 4 (2011) 1369–1375, https://doi.10.1016/j.jmbbm.2011.05.007.
[15] H. Nosrati, R. Aramideh Khouy, A. Nosrati, M. Khodaei, M. Banitalebi-Dehkordi,
K. Ashrafi-Dehkordi, S. Sanami, Z. Alizadeh, Nanocomposite scaffolds for
accelerating chronic wound healing by enhancing angiogenesis,
J. Nanobiotechnol. 19 (2021) 1. 〈https://doi.10.1186/s12951-020-00755-7〉.
[16] A. Bignon, J. Chouteau, J. Chevalier, G. Fantozzi, J.-P. Carret, P.P. Chavassiux,
G. Boivin, M. Melin, D. Hartmann, Effect of micro- and macroporosity of bone
substitutes on their mechanical properties and cellular response, J. Mater. Sci.
Mater. Med. 14 (2003) 1089–1097. 〈https://doi.10.1023/B:JMSM.000000400
6.90399.b4〉.
[17] K.F. Leong, C.K. Chua, N. Sudarmadji, W.Y. Yeong, Engineering functionally
graded tissue engineering scaffolds, J. Mech. Behav. Biomed. Mater. 1 (2008)
140–152, https://doi.10.1016/j.jmbbm.2007.11.002.
[18] I.T. Ozbolat, A.K.M.B. Khoda, M. Marchany, J.A. Gardella, B. Koc, Hybrid tissue
scaffolds for controlled release applications, Virtual Phys. Prototyp. 7 (2012)
37–47. 〈https://doi.10.1080/17452759.2012.668700〉.
[19] R.F. Diegelmann, M.C. Evans, Wound healing: an overview of acute, fibrotic and
delayed healing, Front. Biosci. 9 (2004) 283–289. 〈https://doi.10.2741/1184〉.
[20] K. Deonarine, M.C. Panelli, M.E. Stashower, P. Jin, K. Smith, H.B. Slade,
C. Norwood, E. Wang, F.M. Marincola, D.F. Stroncek, Gene expression profiling of
cutaneous wound healing, J. Transl. Med 5 (2007) 11. 〈https://doi.10.1186/1
479-5876-5-11〉.
[21] K. Garg, N.A. Pullen, C.A. Oskeritzian, J.J. Ryan, G.L. Bowlin, Macrophage
functional polarization (M1/M2) in response to varying fiber and pore
dimensions of electrospun scaffolds, Biomaterials 34 (2013) 4439–4451.
〈https://doi.10.1016/j.biomaterials.2013.02.065〉.
[22] X. Zhu, W. Cui, X. L, Y. Jin, Electrospun fibrous mats with high porosity as
potential scaffolds for skin tissue engineering, Biomacromolecules 9 (2008)
1795–1801. 〈https://doi.10.1021/bm800476u〉.
[23] J. Reignier, M.A. Huneault, Preparation of interconnected poly(ε-caprolactone)
porous scaffolds by a combination of polymer and salt particulate leaching,
Polymer 47 (2006) 4703–4717, https://doi.10.1016/j.polymer.2006.04.029.
Z. Zhang et al.
[24] A.G. Mikos, J.S. Temenoff, Formation of highly porous biodegradable scaffolds
for tissue engineering, Electron. J. Biotechnol. 3 (2000) 23–24. 〈https://doi.10.40
67/S0717-34582000000200003〉.
[25] S. Mahboudi, M. Pezeshki-Modaress, K.A. Noghabi, The study of fibroblast cell
growth on the porous scaffold of gelatin–starch blend using the salt-leaching and
lyophilization method, Int. J. Polym. Mater. Polym. Biomater. 64 (2015)
653–659. 〈https://doi.10.1080/00914037.2014.1002095〉.
[26] H. Amir Afshar, A. Ghaee, Preparation of aminated chitosan/alginate scaffold
containing halloysite nanotubes with improved cell attachment, Carbohydr.
Polym. 151 (2016) 1120–1131, https://doi.10.1016/j.carbpol.2016.06.063.
[27] S.A. Poursamar, J. Hatami, A.N. Lehner, C.L. da Silva, F.C. Ferreira, A.P.
M. Antunes, Potential application of gelatin scaffolds prepared throughin situgas
foaming in skin tissue engineering, Int. J. Polym. Mater. Polym. Biomater. 65
(2016) 315–322. 〈https://doi.10.1080/00914037.2015.1119688〉.
[28] Y.R. Park, H.W. Ju, J.M. Lee, D.K. Kim, O.J. Lee, B.M. Moon, H.J. Park, J.
Y. Jeong, Y.K. Yeon, C.H. Park, Three-dimensional electrospun silk-fibroin
nanofiber for skin tissue engineering, Int J. Biol. Macromol. 93 (2016)
1567–1574, https://doi.10.1016/j.ijbiomac.2016.07.047.
[29] B.S. Kim, Y.W. Kwon, J.S. Kong, G.T. Park, G. Gao, W. Han, M.B. Kim, H. Lee, J.
H. Kim, D.W. Cho, 3D cell printing of in vitro stabilized skin model and in vivo
pre-vascularized skin patch using tissue-specific extracellular matrix bioink: a
step towards advanced skin tissue engineering, Biomaterials 168 (2018) 38–53,
https://doi.10.1016/j.biomaterials.2018.03.040.
[30] N.R. Eviana Putri, X. Wang, Y. Chen, X. Li, N. Kawazoe, G. Chen, Preparation of
PLGA-collagen hybrid scaffolds with controlled pore structures for cartilage tissue
engineering, Prog. Nat. Sci. Mater. Int. 30 (2020) 642–650, https://doi.10.1016/
j.pnsc.2020.07.003.
[31] J. Zhang, H. Zhang, L. Wu, J. Ding, Fabrication of three dimensional polymeric
scaffolds with spherical pores, J. Mater. Sci. 41 (2006) 1725–1731. 〈https://do
i.10.1007/s10853-006-2873-7〉.
[32] Q. Hou, D.W. Grijpma, J. Feijen, Porous polymeric structures for tissue
engineering prepared by a coagulation, compression moulding and salt leaching
technique, Biomaterials 24 (2003) 1937–1947, https://doi.10.1016/s0142-9612
(02)00562-8.
[33] Q. Zhang, H. Lu, N. Kawazoe, G. Chen, Pore size effect of collagen scaffolds on
cartilage regeneration, Acta Biomater. 10 (2014) 2005–2013, https://
doi.10.1016/j.actbio.2013.12.042.
[34] A. Madni, R. Khan, M. Ikram, S.S. Naz, T. Khan, F. Wahid, Fabrication and
characterization of chitosan–vitamin C–lactic acid composite membrane for
potential skin tissue engineering, Int. J. Polym. Sci. 2019 (2019) 1–8. 〈https
://doi.10.1155/2019/4362395〉.
[35] M.G.N. Campos, L.H.I. Mei, A.R. Santos Jr., Sorbitol-plasticized and neutralized
chitosan membranes as skin substitutes, Mater. Res. 18 (2015) 781–790.
〈https://doi.10.1590/1516-1439.025015〉.
[36] A.G. Mikos, A.J. Thorsen, L.A. Czerwonka, Y. Bao, R. Langer, Preparation and
characterization of poly(L-lactic acid) foams, Polymer 35 (1994) 1068–1077,
https://doi.10.1016/0032-3861(94)90953-9.
[37] D. Sin, X. Miao, G. Liu, F. Wei, G. Chadwick, C. Yan, T. Friis, Polyurethane (PU)
scaffolds prepared by solvent casting/particulate leaching (SCPL) combined with
centrifugation, Mater. Sci. Eng.: C. 30 (2010) 78–85, https://doi.10.1016/j.
msec.2009.09.002.
[38] J. Chen, J. Ye, X. Liao, S. Li, W. Xiao, Q. Yang, G. Li, Organic solvent free
preparation of porous scaffolds based on the phase morphology control using
supercritical CO2, J. Supercrit. Fluids 149 (2019) 88–96, https://doi.10.1016/j.
supflu.2019.03.021.
[39] M. Flaibani, N. Elvassore, Gas anti-solvent precipitation assisted salt leaching for
generation of micro- and nano-porous wall in bio-polymeric 3D scaffolds, Mater.
Sci. Eng. C. Mater. Biol. Appl. 32 (2012) 1632–1639, https://doi.10.1016/j.
msec.2012.04.054.
[40] O. Catanzano, V. D’Esposito, P. Formisano, J.S. Boateng, F. Quaglia, Composite
alginate-hyaluronan sponges for the delivery of tranexamic acid in postextractive
alveolar wounds, J. Pharm. Sci. 107 (2018) 654–661, https://doi.10.1016/j.
xphs.2017.09.026.
[41] S. Deville, E. Saiz, R.K. Nalla, A.P. Tomsia, Freezing as a path to build complex
composites, Science 311 (2006) 515–518, https://doi.10.1126/science.1120937.
[42] K.M. Pawelec, A. Husmann, S.M. Best, R.E. Cameron, Understanding anisotropy
and architecture in ice-templated biopolymer scaffolds, Mater. Sci. Eng. C. Mater.
Biol. Appl. 37 (2014) 141–147. 〈https://doi.10.1016/j.msec.2014.01.009〉.
[43] M.B. Frank, S.E. Naleway, T. Haroush, C.H. Liu, S.H. Siu, J. Ng, I. Torres,
A. Ismail, K. Karandikar, M.M. Porter, O.A. Graeve, J. McKittrick, Stiff, porous
scaffolds from magnetized alumina particles aligned by magnetic freeze casting,
Mater. Sci. Eng. C. Mater. Biol. Appl. 77 (2017) 484–492, https://doi.10.1016/j.
msec.2017.03.246.
[44] R. Ghafari, M. Jonoobi, L.M. Amirabad, K. Oksman, A.R. Taheri, Fabrication and
characterization of novel bilayer scaffold from nanocellulose based aerogel for
skin tissue engineering applications, Int J. Biol. Macromol. 136 (2019) 796–803,
https://doi.10.1016/j.ijbiomac.2019.06.104.
[45] H. Yu, X. Chen, J. Cai, D. Ye, Y. Wu, L. Fan, P. Liu, Novel porous three-
dimensional nanofibrous scaffolds for accelerating wound healing, Chem. Eng. J.
369 (2019) 253–262, https://doi.10.1016/j.cej.2019.03.091.
[46] L.S. Wray, J. Rnjak-Kovacina, B.B. Mandal, D.F. Schmidt, E.S. Gil, D.L. Kaplan,
A silk-based scaffold platform with tunable architecture for engineering critically-
sized tissue constructs, Biomaterials 33 (2012) 9214–9224, https://doi.10.1016/
j.biomaterials.2012.09.017.
[47] C. Lloyd, J. Besse, S. Boyce, Controlled-rate freezing to regulate the structure of
collagen-glycosaminoglycan scaffolds in engineered skin substitutes, J. Biomed.
16
Materials Today Communications 32 (2022) 104109
Mater. Res B Appl. Biomater. 103 (2015) 832–840. 〈https://doi.10.1002/jbm.b.
33253〉.
[48] M.W. Pot, K.A. Faraj, A. Adawy, W.J. van Enckevort, H.T. van Moerkerk, E. Vlieg,
W.F. Daamen, T.H. van Kuppevelt, Versatile wedge-based system for the
construction of unidirectional collagen scaffolds by directional freezing: practical
and theoretical considerations, ACS Appl. Mater. Interfaces 7 (2015) 8495–8505.
〈https://doi.10.1021/acsami.5b00169〉.
[49] Y. Wang, X. Wang, J. Shi, R. Zhu, J. Zhang, Z. Zhang, D. Ma, Y. Hou, F. Lin,
J. Yang, M. Mizuno, A biomimetic silk fibroin/sodium alginate composite scaffold
for soft tissue engineering, Sci. Rep. 6 (2016) 39477. 〈https://doi.10.1038/sr
ep39477〉.
[50] S. Fernandes-Silva, J. Moreira-Silva, T.H. Silva, R.I. Perez-Martin, C.G. Sotelo, J.
F. Mano, A.R. Duarte, R.L. Reis, Porous hydrogels from shark skin collagen
crosslinked under dense carbon dioxide atmosphere, Macromol. Biosci. 13 (2013)
1621–1631. 〈https://doi.10.1002/mabi.201300228〉.
[51] W. Yang, H. Xu, Y. Lan, Q. Zhu, Y. Liu, S. Huang, S. Shi, A. Hancharou, B. Tang,
R. Guo, Preparation and characterisation of a novel silk fibroin/hyaluronic acid/
sodium alginate scaffold for skin repair, Int J. Biol. Macromol. 130 (2019) 58–67,
https://doi.10.1016/j.ijbiomac.2019.02.120.
[52] L. Ran, Y. Zou, J. Cheng, F. Lu, Silver nanoparticles in situ synthesized by
polysaccharides from Sanghuangporus sanghuang and composites with chitosan
to prepare scaffolds for the regeneration of infected full-thickness skin defects, Int
J. Biol. Macromol. 125 (2018) 392–403, https://doi.10.1016/j.
ijbiomac.2018.12.052.
[53] J. Yuan, Q. Hou, D. Chen, L. Zhong, X. Dai, Z. Zhu, M. Li, X. Fu, Chitosan/LiCl
composite scaffolds promote skin regeneration in full-thickness loss, Sci. China
Life Sci. 63 (2019) 552–562. 〈https://doi.10.1007/s11427-018-9389-6〉.
[54] F. Han, Y. Dong, Z. Su, R. Yin, A. Song, S. Li, Preparation, characteristics and
assessment of a novel gelatin-chitosan sponge scaffold as skin tissue engineering
material, Int J. Pharm. 476 (2014) 124–133, https://doi.10.1016/j.
ijpharm.2014.09.036.
[55] C. Okolieocha, D. Raps, K. Subramaniam, V. Altstädt, Microcellular to
nanocellular polymer foams: Progress (2004–2015) and future directions – a
review, Eur. Polym. J. 73 (2015) 500–519, https://doi.10.1016/j.
eurpolymj.2015.11.001.
[56] J. Jiang, M.A. Carlson, M.J. Teusink, H. Wang, M.R. MacEwan, J. Xie, Expanding
two-dimensional electrospun nanofiber membranes in the third dimension by a
modified gas-foaming technique, ACS Biomater. Sci. Eng. 1 (2015) 991–1001.
〈https://doi.10.1021/acsbiomaterials.5b00238〉.
[57] J. Jiang, S. Chen, H. Wang, M.A. Carlson, A.F. Gombart, J. Xie, CO2-expanded
nanofiber scaffolds maintain activity of encapsulated bioactive materials and
promote cellular infiltration and positive host response, Acta Biomater. 68 (2018)
237–248, https://doi.10.1016/j.actbio.2017.12.018.
[58] K. Zhang, X. Bai, Z. Yuan, X. Cao, X. Jiao, Y. Li, Y. Qin, Y. Wen, X. Zhang, Layered
nanofiber sponge with an improved capacity for promoting blood coagulation
and wound healing, Biomaterials 204 (2019) 70–79, https://doi.10.1016/j.
biomaterials.2019.03.008.
[59] S.K. Goel, E.J. Beckmn, Generation of microcellular polymeric foams using
supercritical carbon dioxide. II Cell growth and skin formation, Polym. Eng. Sci.
34 (1994) 1148–1156. 〈https://doi.10.1002/pen.760341408〉.
[60] B. Krause, H. Sijbesma, P. Münüklü, N. Vegt, M. Wessling, Bicontinuous
nanoporous polymers by carbon dioxide foaming, Macromolecules 34 (2001)
8792–8801. 〈https://doi.10.1021/ma010854j〉.
[61] R. Sanz-Horta, E. Martinez-Campos, C. García, H. Reinecke, A. Gallardo,
J. Rodriguez-Hernandez, C. Elvira, Breath figures makes porous the “so-called”
skin layer obtained in polymer foams prepared by supercritical CO2 treatments,
J. Supercrit. Fluids 167 (2021), 105051 https://doi.10.1016/j.
supflu.2020.105051.
[62] I. Manavitehrani, T.Y.L. Le, S. Daly, Y. Wang, P.K. Maitz, A. Schindeler,
F. Dehghani, Formation of porous biodegradable scaffolds based on poly
(propylene carbonate) using gas foaming technology, Mater. Sci. Eng. C Mater.
Biol. Appl. 96 (2019) 824–830, https://doi.10.1016/j.msec.2018.11.088.
[63] Q. Ren, M. Wu, W. Li, X. Zhu, Y. Zhao, L. Wang, W. Zheng, A green fabrication
method of poly (lactic acid) perforated membrane via tuned crystallization and
gas diffusion process, Int J. Biol. Macromol. 182 (2021) 1037–1046, https://
doi.10.1016/j.ijbiomac.2021.04.105.
[64] Y. Chen, Z. Jia, M. Shafiq, X. Xie, X. Xiao, R. Castro, J. Rodrigues, J. Wu, G. Zhou,
X. Mo, Gas foaming of electrospun poly(L-lactide-co-caprolactone)/silk fibroin
nanofiber scaffolds to promote cellular infiltration and tissue regeneration,
Colloids Surf. B Biointerfaces 201 (2021), 111637 https://doi.10.1016/j.
colsurfb.2021.111637.
[65] M. Hajiabbas, I. Alemzadeh, M. Vossoughi, A porous hydrogel-electrospun
composite scaffold made of oxidized alginate/gelatin/silk fibroin for tissue
engineering application, Carbohydr. Polym. 245 (2020), 116465 https://
doi.10.1016/j.carbpol.2020.116465.
[66] X. Jing, H. Li, H.-Y. Mi, Y.-J. Liu, Y.-M. Tan, Fabrication of fluffy shish-kebab
structured nanofibers by electrospinning, CO2 escaping foaming and controlled
crystallization for biomimetic tissue engineering scaffolds, Chem. Eng. J. 372
(2019) 785–795, https://doi.10.1016/j.cej.2019.04.194.
[67] B. Balusamy, A. Senthamizhan, T. Uyar, Electrospun nanofibrous materials for
wound healing applications, Electro Mater. Tissue Eng. Biomed. Appl. (2017)
147–177.
[68] S.P. Miguel, R.S. Sequeira, A.F. Moreira, C.S.D. Cabral, A.G. Mendonca,
P. Ferreira, I.J. Correia, An overview of electrospun membranes loaded with
bioactive molecules for improving the wound healing process, Eur. J. Pharm.
Biopharm. 139 (2019) 1–22, https://doi.10.1016/j.ejpb.2019.03.010.
Z. Zhang et al.
[69] C.E. Ayres, B.S. Jha, S.A. Sell, G.L. Bowlin, D.G. Simpson, Nanotechnology in the
design of soft tissue scaffolds: innovations in structure and function, WIREs
Nanomed. Nanobiotechnol. 2 (2010) 20–34. 〈https://doi.10.1002/wnan.055〉.
[70] A. Haider, S. Haider, I.-K. Kang, A comprehensive review summarizing the effect
of electrospinning parameters and potential applications of nanofibers in
biomedical and biotechnology, Arab. J. Chem. 11 (2018) 1165–1188, https://
doi.10.1016/j.arabjc.2015.11.015.
[71] Y. Zhang, H. Ouyang, C.T. Lim, S. Ramakrishna, Z.M. Huang, Electrospinning of
gelatin fibers and gelatin/PCL composite fibrous scaffolds, J. Biomed. Mater. Res
B Appl. Biomater. 72 (2005) 156–165. 〈https://doi.10.1002/jbm.b.30128〉.
[72] K. Tuzlakoglu, N. Bolgen, A.J. Salgado, M.E. Gomes, E. Piskin, R.L. Reis, Nano-
and micro-fiber combined scaffolds A new architecture for bone tissue
engineering, J. Mater. Sci. Mater. Med. 16 (2005) 1099–1104. 〈https://do
i.10.1007/s10856-005-4713-8〉.
[73] L. Huang, J. Huang, H. Shao, X. Hu, C. Cao, S. Fan, L. Song, Y. Zhang, Silk
scaffolds with gradient pore structure and improved cell infiltration performance,
Mater. Sci. Eng. C. Mater. Biol. Appl. 94 (2019) 179–189, https://doi.10.1016/j.
msec.2018.09.034.
[74] E. Zhmayev, H. Zhou, Y.L. Joo, Modeling of non-isothermal polymer jets in melt
electrospinning, J. Non Newton. Fluid Mech. 153 (2008) 95–108, https://
doi.10.1016/j.jnnfm.2007.11.011.
[75] G. Hochleitner, T. Jungst, T.D. Brown, K. Hahn, C. Moseke, F. Jakob, P.D. Dalton,
J. Groll, Additive manufacturing of scaffolds with sub-micron filaments via melt
electrospinning writing, Biofabrication 7 (2015), 035002. 〈https://doi.10.1088/1
758-5090/7/3/035002〉.
[76] G. Hochleitner, F. Chen, C. Blum, P.D. Dalton, B. Amsden, J. Groll, Melt
electrowriting below the critical translation speed to fabricate crimped elastomer
scaffolds with non-linear extension behaviour mimicking that of ligaments and
tendons, Acta Biomater. 72 (2018) 110–120, https://doi.10.1016/j.
actbio.2018.03.023.
[77] A. Hrynevich, B.S. Elci, J.N. Haigh, R. McMaster, A. Youssef, C. Blum, T. Blunk,
G. Hochleitner, J. Groll, P.D. Dalton, Dimension-Based Design of Melt
Electrowritten Scaffolds, Small 14 (2018), e1800232. 〈https://doi.10.1002/
smll.201800232〉.
[78] T.D. Brown, P.D. Dalton, D.W. Hutmacher, Melt electrospinning today: an
opportune time for an emerging polymer process, Prog. Polym. Sci. 56 (2016)
116–166, https://doi.10.1016/j.progpolymsci.2016.01.001.
[79] T.D. Brown, A. Slotosch, L. Thibaudeau, A. Taubenberger, D. Loessner,
C. Vaquette, P.D. Dalton, D.W. Hutmacher, Design and fabrication of tubular
scaffolds via direct writing in a melt electrospinning mode, Biointerphases 7
(2012) 13. 〈https://doi.10.1007/s13758-011-0013-7〉.
[80] N. Abbasi, A. Abdal-Hay, S. Hamlet, E. Graham, S. Ivanovski, Effects of Gradient
and Offset Architectures on the Mechanical and Biological Properties of 3-D Melt
Electrowritten (MEW) Scaffolds, ACS Biomater. Sci. Eng. 5 (2019) 3448–3461.
〈https://doi.10.1021/acsbiomaterials.8b01456〉.
[81] N.T. Nguyen, J.H. Kim, Y.H. Jeong, Identification of sagging in melt-
electrospinning of microfiber scaffolds, Mater. Sci. Eng. C Mater. Biol. Appl. 103
(2019), 109785 https://doi.10.1016/j.msec.2019.109785.
[82] T.D. Brown, F. Edin, N. Detta, A.D. Skelton, D.W. Hutmacher, P.D. Dalton, Melt
electrospinning of poly(epsilon-caprolactone) scaffolds: phenomenological
observations associated with collection and direct writing, Mater. Sci. Eng. C
Mater. Biol. Appl. 45 (2014) 698–708, https://doi.10.1016/j.msec.2014.07.034.
[83] P.D. Dalton, Melt electrowriting with additive manufacturing principles, Curr.
Opin. Biomed. Eng. 2 (2017) 49–57, https://doi.10.1016/j.cobme.2017.05.007.
[84] L. Koch, S. Kuhn, H. Sorg, M. Gruene, S. Schlie, R. Gaebel, B. Polchow,
K. Reimers, S. Stoelting, N. Ma, Laser printing of skin cells and human stem cells,
Tissue Eng. Part C Methods 16 (2010) 847–854. 〈https://doi.10.1089/ten.
TEC.2009.0397〉.
[85] L. Koch, A. Deiwick, S. Schlie, S. Michael, M. Gruene, V. Coger, D. Zychlinski,
A. Schambach, K. Reimers, P.M. Vogt, B. Chichkov, Skin tissue generation by laser
cell printing, Biotechnol. Bioeng. 109 (2012) 1855–1863. 〈https://doi.10.
1002/bit.24455〉.
[86] A. Bigham, F. Foroughi, E. Rezvani Ghomi, M. Rafienia, R.E. Neisiany,
S. Ramakrishna, The journey of multifunctional bone scaffolds fabricated from
traditional toward modern techniques, Bio Des. Manuf. 3 (2020) 281–306. 〈https
://doi.10.1007/s42242-020-00094-4〉.
[87] J. Malda, J. Visser, F.P. Melchels, T. Jungst, W.E. Hennink, W.J. Dhert, J. Groll, D.
W. Hutmacher, 25th anniversary article: engineering hydrogels for
biofabrication, Adv. Mater. 25 (2013) 5011–5028, https://doi.10.1002/
adma.201302042.
[88] X. Zhang, Y. Zhang, Tissue engineering applications of three-dimensional
bioprinting, Cell Biochem Biophys. 72 (2015) 777–782, https://doi.10.1007/
s12013-015-0531-x.
[89] B. Zhang, Y. Luo, L. Ma, L. Gao, Y. Li, Q. Xue, H. Yang, Z. Cui, 3D bioprinting an
emerging technology full of opportunities and challenges, Bio Des. Manuf. 1
(2018) 2–13. 〈https://doi.10.1007/s42242-018-0004-3〉.
17
Materials Today Communications 32 (2022) 104109
[90] C. Colosi, S.R. Shin, V. Manoharan, S. Massa, M. Costantini, A. Barbetta, M.
R. Dokmeci, M. Dentini, A. Khademhosseini, Microfluidic bioprinting of
heterogeneous 3D tissue constructs using low-viscosity bioink, Adv. Mater. 28
(2016) 677–684. 〈https://doi.10.1002/adma.201503310〉.
[91] A. Skardal, D. Mack, E. Kapetanovic, A. Atala, J.D. Jackson, J. Yoo, S. Soker,
Bioprinted amniotic fluid-derived stem cells accelerate healing of large skin
wounds, Stem Cells Transl. Med 1 (2012) 792–802. 〈https://doi.10.5966/sctm.20
12-0088〉.
[92] H.W. Kang, S.J. Lee, I.K. Ko, C. Kengla, J.J. Yoo, A. Atala, A 3D bioprinting system
to produce human-scale tissue constructs with structural integrity, Nat.
Biotechnol. 34 (2016) 312–319. 〈https://doi.10.1038/nbt.3413〉.
[93] J. Li, C. Wu, P.K. Chu, M. Gelinsky, 3D printing of hydrogels: rational design
strategies and emerging biomedical applications, Mater. Sci. Eng.: R: Rep. 140
(2020), 100543 https://doi.10.1016/j.mser.2020.100543.
[94] S. Vijayavenkataraman, W.C. Yan, W.F. Lu, C.H. Wang, J.Y.H. Fuh, 3D
bioprinting of tissues and organs for regenerative medicine, Adv. Drug Deliv. Rev.
132 (2018) 296–332, https://doi.10.1016/j.addr.2018.07.004.
[95] N. Cubo, M. Garcia, J.F. Del Canizo, D. Velasco, J.L. Jorcano, 3D bioprinting of
functional human skin: production and in vivo analysis, Biofabrication 9 (2016),
015006. 〈https://doi.10.1088/1758-5090/9/1/015006〉.
[96] J. Fuh, Micro- and bio-rapid prototyping using drop-on-demand 3D printing,
Handb. Manuf. Eng. Technol. (2013) 1–15.
[97] H. Tseng, J.A. Gage, W.L. Haisler, S.K. Neeley, T. Shen, C. Hebel, H.G. Barthlow,
M. Wagoner, G.R. Souza, A high-throughput in vitro ring assay for vasoactivity
using magnetic 3D bioprinting, Sci. Rep. 6 (2016) 30640. 〈https://doi.10.1038/sr
ep30640〉.
[98] F. Zhou, Y. Hong, R. Liang, X. Zhang, Y. Liao, D. Jiang, J. Zhang, Z. Sheng, C. Xie,
Z. Peng, X. Zhuang, V. Bunpetch, Y. Zou, W. Huang, Q. Zhang, E.V. Alakpa,
S. Zhang, H. Ouyang, Rapid printing of bio-inspired 3D tissue constructs for skin
regeneration, Biomaterials 258 (2020), 120287 https://doi.10.1016/j.
biomaterials.2020.120287.
[99] C. Niu, L. Wang, D. Ji, M. Ren, D. Ke, Q. Fu, K. Zhang, X. Yang, Fabrication of SA/
Gel/C scaffold with 3D bioprinting to generate micro-nano porosity structure for
skin wound healing: a detailed animal in vivo study, Cell Regen. 11 (2022) 10.
〈https://doi.10.1186/s13619-022-00113-y〉.
[100] A. De Mori, M. Pena Fernandez, G. Blunn, G. Tozzi, M. Roldo, 3D printing and
electrospinning of composite hydrogels for cartilage and bone tissue engineering,
Polymers 10 (2018) 285. 〈https://doi.10.3390/polym10030285〉.
[101] K.J. De France, F. Xu, T. Hoare, Structured macroporous hydrogels: progress,
challenges, and opportunities, Adv. Health Mater. 7 (2018) 1700927. 〈https://
doi.10.1002/adhm.201700927〉.
[102] X. Liu, M. Hao, Z. Chen, T. Zhang, J. Huang, J. Dai, Z. Zhang, 3D bioprinted
neural tissue constructs for spinal cord injury repair, Biomaterials 272 (2021),
120771 https://doi.10.1016/j.biomaterials.2021.120771.
[103] X. Zhou, Y. Feng, J. Zhang, Y. Shi, L. Wang, Recent advances in additive
manufacturing technology for bone tissue engineering scaffolds, The, Int. J. Adv.
Manuf. Technol. 108 (2020) 3591–3606. 〈https://doi.10.1007/s00170-020-
05444-1〉.
[104] L. Shao, Q. Gao, C. Xie, J. Fu, M. Xiang, Z. Liu, L. Xiang, Y. He, Sacrificial
microgel-laden bioink-enabled 3D bioprinting of mesoscale pore networks, Bio
Des. Manuf. 3 (2020) 30–39. 〈https://doi.10.1007/s42242-020-00062-y〉.
[105] S. Hasan, K. Hamersveld, M. Mheen, B.L. Kaptein, T. Larsen, Migration of a novel
3D-printed cementless versus a cemented total knee arthroplasty: Two-year
results of a randomized controlled trial using radiostereometric analysis, BONE Jt.
J. 102-B (2020) 1016–1024. 〈https://doi.10.1302/0301-620X.102B8〉.
[106] V. Patel, D. Kovalsky, S.C. Meyer, A. Chowdhary, H. Lockstadt, F. Techy,
C. Langel, R. Limoni, P.S. Yuan, A. Kranenburg, D. Cher, G. Tender, T.J. Hillen,
Prospective trial of sacroiliac joint fusion using 3D-printed triangular titanium
implants, Med Devices (Auckl. ) 13 (2020) 173–182. 〈https://doi.10.
2147/MDER.S253741〉.
[107] F.J. O’Brien, B.A. Harley, I.V. Yannas, L.J. Gibson, The effect of pore size on cell
adhesion in collagen-GAG scaffolds, Biomaterials 26 (2005) 433–441, https://
doi.10.1016/j.biomaterials.2004.02.052.
[108] H. Janik, M. Marzec, A review: fabrication of porous polyurethane scaffolds,
Mater. Sci. Eng. C. Mater. Biol. Appl. 48 (2015) 586–591, https://doi.10.1016/j.
msec.2014.12.037.
[109] T.J. Sill, H.A. von Recum, Electrospinning: applications in drug delivery and
tissue engineering, Biomaterials 29 (2008) 1989–2006, https://doi.10.1016/j.
biomaterials.2008.01.011.
[110] Y. Liang, Y. Liang, H. Zhang, B. Guo, Antibacterial biomaterials for skin wound
dressing, Asian J. Pharm. Sci. 17 (2022) 353–384, https://doi.10.1016/j.
ajps.2022.01.001.
[111] A. Olejnik, J.A. Semba, A. Kulpa, A. Danczak-Pazdrowska, J.D. Rybka,
J. Gornowicz-Porowska, 3D bioprinting in skin related research: recent
achievements and application perspectives, ACS Synth. Biol. 11 (2022) 26–38.
〈https://doi.10.1021/acssynbio.1c00547〉.