In AA, Fibroblast Wound to Wrinkles:

Growth Factor Emerging treatment

Can Be A Companion options of neuropeptide
of Topical Minoxidil

Pg. 2 Pg. 3

Vol. No. 19 Pune, June 2021

Physiological Changes in Young and Old Woman Scalp

In women, aging leads to reduced hair density and
thinner fibres and can result in femalepattern hair loss

(a, b) Representative images of age-related changes in female scalp

skin showing flattening of the dermal‒epidermal junction and loss
of rete ridges and structural changes in the papillary dermis and the
organization of dermal collagen. (a) Scalp of a female aged 20 years.
Bar ¼ 200 mm. (b) Scalp of a female aged 81 years. Bar = 200 mm.
(c) Quantification of age-related changes in epidermal area in female
scalp skin

Schematic of age-related changes in the female scalp dermal

environment and the potential impact on residing HFs. Structural
and biochemical changes take place in the aging scalp, including
delineation of the papillary dermis, loss of rete ridges, and
organization of collagen.

Low-grade inflammation due to increased ROS and solar elastosis

may initiate the evacuation of DS cells from the HF to restore the
impaired dermis, leading to a reduced capacity for DS contraction
and replenishment of the DP, impeding the hair cycle, and resulting
in smaller HFs. Dermal fibrosis would also impede remodeling of
the lower portion of the HF and reduce its capacity to move down
into the hypodermis. DP, dermal papilla; DS, dermal sheath; dWAT,
dermal white adipose tissue; HF, hair follicle.

In the female scalp, the dermis has to support large, predominately

anagen HFs cycling in an asynchronous manner. Whether the shift in
scalp DF phenotype with aging is due to a loss in cellular identity or
the evacuation of DS cells at the expense of the HF to support the
aging interfollicular dermis remains to be established.

Further studies on aging human DFs from nonhaired body sites may
help to clarify this. In summary, with aging, the female scalp shows
striking structural and biological changes in the HF environment,
including a shift in the DF phenotype, rendering it unable to fully
support hair cycle remodeling and leading to an aged
hair phenotype.

HAS2, hyaluronic acid synthase 2; MMP1, Ref. Journal of Investigative Dermatology (2021) 141, 1041e1051;
doi:10.1016/j.jid.2020.11.009
matrix metalloproteinase-1
Vol. No. 19 Pg. No. 2

In AA, Fibroblast Growth Factor Can Be A Companion of Topical Minoxidil

A study was conducted to compare the safety and efficacy of topical Hair density among groups at baseline &
minoxidil, nano-microneedle-assisted Fibroblast Growth Factor after 16-week treatment
(FGF), and a combination of the two. In this study the participants
suffering from male androgenetic alopecia were divided in 3 groups,
topical minoxidil, nano-microneedle-assisted FGF, and a
combination of the two.
In this study 40 male in the age group of 22 to 50 years diagnosed
with male androgenetic alopecia (MAA) were enrolled. They did
not undergo any treatment affecting the hair cycle within the 6
months before enrollment. Those with other hair disorders or known
systemic diseases were excluded.
Participants were randomized into four groups of 10 each using the
random number table method. Details of the groups are as follows:

Hair density increased in group F but not in group S. This suggests that the mechanical
stimulus of the nano-microneedle does not produce hair growth but it promotes the
penetration of FGF. Hair density was also higher in group MF than that in groups M and F
(both P < 0.05), indicating that combination treatment is superior to individual treatments.

Assessment of patients

No change Better
Therapy was administered for 16 weeks. 12
Saline (group S) and FGF (groups F and MF) were applied once 10
9 10 9
weekly using a nano-microneedle. After spraying 1 mL of saline or
Number of patients

8
FGF on the balding region, the nano-microneedle was moved 8

smoothly and slowly on the scalp. Topical minoxidil (groups M and 6

MF, seven drops per treatment) was applied to the balding region
4
every morning and evening, followed by local massage for 5 to 10 2
minutes. 2 1 1
Patients were evaluated monthly for safety and efficacy. At each 0
0
visit, standardized global photographs were obtained using a digital Group S Group M Group F Group MF
camera.
At the end of the 16-week treatment period, participants and two
dermatologists blinded to the treatment evaluated the treatment Assessment of investigators
efficacy based on the photographs taken at baseline and at 16 weeks.
The assessment was measured on a six-point scale: _1 = worsened, 0 No change Better
= unchanged, 1 = slight improvement, 2 = moderate improvement, 3 9
10
= good improvement, and 4 = excellent improvement.
Number of patients

8
8 7
6
6
4
4 3
2
2 1
0
Group S Group M Group F Group MF

No serious adverse effects were encountered during the treatment

period. Pain during nano-microneedle treatment was well tolerated
by patients in groups S, F, and MF.

In the present study, we found that hair density of patients receiving

Photographs of patients' hair at baseline (left) and after the 16-week treatment (right). (A,
B): Nano-microneedle-assisted normal saline. (C, D) 5% topical minoxidil. (E, F):
Nanomicroneedle- assisted fibroblast growth factor. (G, H): 5% topical minoxidil and
nano-microneedle-assisted fibroblast growth factor. Photographs showing hair diameter at
baseline (left) and after 16-week treatment (right) using a hair microscope. (I, J): Nano-
microneedle-assisted saline. (K, L): 5% topical minoxidil. (M, N): Nano-microneedle
assisted fibroblast growth factor. (O, P): Topical minoxidil and nano-microneedle-assisted
fibroblast growth factor.
minoxidil, nano-microneedle-assisted FGF, or both increased
notably over baseline, with patients treated using combined
treatments achieving the most satisfactory results.
Thus, the combination of nano-microneedle-assisted FGF and
topical minoxidil is safe and reliable for treatment of MAA and
better than monotherapy. Further research is required to evaluate the
long-term efficacy and safety of this combination therapy in a larger
group of subjects.
Ref. Combination therapy with topical minoxidil and nanomicroneedle- assisted fibroblast
growth factor for male androgenetic alopecia: a randomized controlled trial in Chinese
patients. Chin Med J 2021;134:851–853. doi: 10.1097/CM9.0000000000001195
Vol. No. 19 Pg. No. 3

Wound to Wrinkles: Emerging treatment options of neuropeptide

Substance P (SP), a small-sized neuropeptide consisting of 11 amino
acids, is produced in wound areas and exhibits effective skin
regenerative activity. The anti-aging effects of SP are attributed to its
anti-inflammatory activity and effects on collagen synthesis. Studies
have shown that SP increases type I procollagen levels, correlated
with the regulation of matrix metalloproteinase (MMP)-1 as well as
anti-inflammatory activity, thus indicating its potential anti-aging
efficacy in clinical practice. Although the potential anti-aging
activity of SP facilitates its use in cosmeceuticals, its application has
been limited owing to its low stability. In a previous study, a stable
SP formulation known as SP-based hydrogel was developed. To
prevent oxidation of SP and minimize physical damage to SP from
an interfacial interaction caused by SP–surfactant association and/or
coating interfaces, sodium thiosulfate and polysorbate 80 were
added to the SP-based hydrogel. In addition, a gelling agent
(hydroxyethyl cellulose) was used to enhance the viscosity and SP
stability of the hydrogel. The SP-based hydrogel demonstrated
superior in vitro stability and anti-aging efficacy compared to SP
alone. In particular, the SP-based hydrogel was stable in growth
medium containing fetal bovine serum for up to 24 h and at high
temperature (~60 °C) for up to 4 weeks. To evaluate SP stability, we
removed aliquots of samples from the growth medium or at high
temperature and then diluted them with phosphate buffered saline
for analysis of SP content by enzyme-linked immunosorbent assay.
Although previous reports on the in vitro anti-aging effects and
better stability profile of SP-based hydrogel indicate the need for
clinical studies screening the anti-aging effects of SP-based
hydrogel, none have been performed to date.
Effect of substance P (SP) gel on the viability of HDFs and in-vitro reconstructed 3D human
skin, keraskin®-FT. (A) For amaging effects of SP gel formulation on cell membrane, HDFs
Skin aging is generally caused by a decline in the components of the were treated with PBS (Con; control), SP alone, or SP gel (1–10 _g/mL) for 24 h, and cell
extracellular matrix (e.g., collagen and elastin) and due to viability was determined by LDH assay. (B,C) Skin irritability was tested using SP gel (1–10
_g/mL) on in-vitro reconstructed 3D human skin, keraskin®-FT. Tissue viability in the SP gel-
inflammatory phenomena. Many growth factors and peptides with treated group was analysed by MTT assay (B) and histological examination (C), PBS was used
cell-growth and collagen-synthesis activities have shown promise in as a control. Values represent mean ± SD from three independent experiments. Each value was
their application in anti-aging materials. However, the effect of compared with the control using Student's t-test (** p < 0.01). Scale bar = 500 _m. HDF, human
dermal fibroblast; LDH, lactate dehydrogenase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-
collagen production, without anti-inflammatory effect, and skin diphenyl tetrazolium bromide; PBS, phosphate-buttered saline; SDS, sodium dodecyl sulfate.
penetration may not be enough for their use in anti-aging agents.
Previously, we reported a substance P (SP)-based hydrogel (SP gel)
that had potential wound-healing activities via induction of skin cell
regeneration and collagen synthesis. Here, we analyzed the anti-
aging activities and skin absorption effects of SP gel to extend its
characterization. Toxicity tests, performed on human dermal
fibroblasts (HDFs) and on a reconstructed 3D human skin model,
indicated SP gel to be safe for long-term use, without causing
irritation, even at high concentrations. In-vitro analysis revealed that
SP gel elicited stronger collagen production activities than SP alone,
and promoted anti-inflammatory effects with increased skin
absorption properties. Moreover, SP gel did not induce melanin
synthesis in a keratinocyte-melanocyte co-culture system. Together,
the results suggest that SP gel has potential cosmetic effects and
applicability as a novel ingredient in anti-aging products.

Effect of SP gel on the production of collagen

To examine the effect of SP gel on collagen synthesis on the skin,
HDFs were treated with SP gel. Our results demonstrated that SP
gel, containing 1–10 _g/mL of SP, significantly (p < 0.01) increased
type I procollagen production to 128.05 _ 5.19, 145.19 _ 6.80, and
150.68 _ 16.70%, compared to the control treatment (PBS) (Figure
2A). Otherwise, as shown in Figure 2A, no noteworthy difference in
type I collagen production was observed across HDFs treated with 1
_g/mL of SP alone. While SP alone did have some effect on collagen
production at concentrations of 5–10 _g/mL (119.96 _ 3.8 and
124.77 _ 8.87%,), this effect was significantly (p < 0.01) lower than
with SP gel
Collagen-increasing effects of SP gel in HDFs. Effects of SP gel on type 1 collagen (A), MMP-1
(B), and TIMP-1 (C) were measured in HDFs. HDFs were treated with PBS (Con; control), SP
alone, or SP gel (1–10 µg/mL) for 24 h and the expression of type I procollagen, MMP-1, and
TIMP-1 was measured using the Procollagen Type I C-peptide (PIP) EIA kit, human MMP-1
ELISA kit, and TIMP-1 ELISA kit, respectively. PBS was used as a control. Values represent the
mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01 vs. PBS-containing
medium without SP; # p < 0.05, ## p < 0.01 vs. SP alone. HDF, human dermal fibroblast; PBS,
phosphate-buffered saline; MMP-1, matrix metalloproteinase-1; TIMP-1, tissue inhibitors of
metalloproteinase-1; ELISA, enzyme-linked immunosorbent assay.
Vol. No. 19 Pg. No. 4

In general, collagen synthesis occurs in HDFs inside the skin. Therefore, based on the previous findings from several studies, SP
Therefore, SP gel should have high skin penetration capacity in gel may be suggested to be absorbed effectively into the skin layer.
order to achieve its anti-aging effects. We, therefore, sought to
examine whether SP gel is able to penetrate into a reconstructed 3D In conclusion, our in-vitro study demonstrated that SP gel treatment
human skin model. After 1 h of topical application, SP gel crossed could increase type I procollagen levels, correlated with the
the SC and accumulated in the epidermis. Little accumulation in the regulation of MMP-1 and TIMP-1, as well as anti-inflammatory
dermal tissue was also observed, suggesting that SP gel can be effect. Moreover, we demonstrated the increased skin absorption
absorbed into the skin. After 24 h, along with accumulation in the capability of SP gel, without causing skin pigmentation.
epidermis, strong green fluorescence was observed in the dermis as Collectively, our findings suggest SP gel to be possibly used as an
well, indicating the presence of SP gel there. Otherwise, after 1 h, effective and safe cosmetic ingredient with anti-aging functions.
weak green fluorescence for SP-only treatment was observed in the
Ref:
dermis, but not after 6 h. This means that SP was degraded in the 1. Rittie, L.; Fisher, G.J. UV-light-induced signal cascades and skin aging. Ageing Res. Rev.
culture conditions of keraskin®-FT, suggesting that the degraded SP 2002, 1, 705–720. [CrossRef]
2. Makrantonaki, E.; Zouboulis, C.C. Molecular mechanisms of skin aging: State of the art.
rapidly penetrated the keraskin®-FT and escaped to the medium Ann. N. Y. Acad. Sci. 2007, 1119, 40–50. [CrossRef] [PubMed]
within 6 h. Although the mechanism by which SP gel gets 3. Pham, Q.L.; Jang, H.J.; Kim, K.B. Antiwrinkle effect of fermented black ginseng on
internalized into the skin is not yet clear, it seems to be related to its human fibroblasts. Int. J. Mol. Med. 2017, 39, 681–686. [CrossRef] [PubMed]
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characteristic molecular structure. Many amphipathic peptides weapons and strategies. Evid.-Based Complement. Altern. Med. 2013, 2013, 827248.
possess skin absorption abilities, which might mean that the [CrossRef] [PubMed]
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amphipathic characteristics of peptides are the major factors for skin of Methylene Blue for Human Skin Longevity. Sci. Rep. 2017, 7, 2475. [CrossRef]
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taken up by mammalian cells via a non-endocytic mechanism. The 6. Pillai, S.; Oresajo, C.; Hayward, J. Ultraviolet radiation and skin aging: Roles of reactive
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7. Ishitsuka, Y.; Maniwa, F.; Koide, C.; Kato, Y.; Nakamura, Y.; Osawa, T.; Tanioka, M.;
diverse mechanisms that include inverted micelle formation, pore Miyachi, Y. Increased halogenated tyrosine levels are useful markers of human skin
formation, the carpet-like model, and the membrane thinning model. ageing, reflecting proteins denatured by past skin inflammation. Clin. Exp. Derm. 2012,
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8. Kim, S.R.; Jung, Y.R.; An, H.J.; Kim, D.H.; Jang, E.J.; Choi, Y.J.; Moon, K.M.; Park,
bind to G-protein coupled receptors, it can be absorbed into the skin. M.H.; Park, C.H.; Chung, K.W.; et al. Anti-wrinkle and anti-inflammatory effects of active
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