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ORIGINAL RESEARCH PAPER
ADULT STEM CELL RELEASED MOLECULES IN COMBINATION WITH
MICRONEEDLING RESTORE HAIR GROWTH
Michael Ryan
Clinics, Dubai & Manchester, UK
Greg Maguire*
NeoGenesis Inc San Diego, CA 92121 USA. *Corresponding Author
INTRODUCTION
Multiple factors contribute to hair loss, including aging, heredity,
hormones, environmental exposure, stress, medications, and nutrition.
Hair growth is a highly complicated stem cell-based process and serves
as a model for general stem cell function where stem cells in the follicle
interact with stem cells and broblasts in the surrounding skin
(Schmidt-Ullrich R and Paus, 2005; Weir and Garza, 2020). The
dermal papilla (DP), a cluster of specialized broblasts in the follicle,
secretes diffusible proteins packaged into exosomes that regulate the
growth and activity of the various cells in the follicle, thereby playing a
key role within the follicle in the regulation of hair cycling and growth
(Zhou et al, 2018). Extracellular vesicles (EVs), including exosomes,
are the carriers for the distribution of morphogens and growth and
differentiation factors (Riazifar et al, 2017), and can pack thousands of
proteins into one exosome, delivering that collective cargo to one
target in the same time and space (Maguire, 2016). The DP is
encapsulated by an overlying matrix of epithelial cells, and growth
factors from DP are believed to cause epithelial cells to proliferate and
differentiate to produce hair shafts during the anagen phase. In addition
to hair loss, Nishimura et al (2005) have demonstrated that hair graying
is caused by defective self-maintenance of melanocyte stem cells in the
follicle. Despite the efforts of scientists in seeking effective therapeutic
agents, only a few marginally effective Food and Drug Administration
(FDA)-approved medications are available for alopecia patients. The
mostly widely prescribed drugs for alopecia are Finasteride and
minoxidil, either as monotherapy or in combination. Although
nasteride has been found to enhance hair growth, oral nasteride
often causes reduced libido, impotence, and sexual dysfunction (Fertig
et al, 2017). Further, this treatment is only applicable to male patients
with AGA given th e highly teratogenic e ffects of nasteride
(Kawashima et al, 2004). Topical minoxidil (2%) is the only treatment
for female patients with AGA, and this has lower efcacy than the 5%
minoxidil preparation that is available for male patients (Olsen et al,
2002), thus resulting in disappointing outcomes. Furthermore,
minoxidil can affect the heart and blood pressure if absorbed
excessively through the skin (Goren and Naccarato, 2018).
While adipose derived mesenchymal stem cells (ADSC) molecules
have been found to regrow hair in a randomized, double blind, placebo
controlled clinical trial (Tak et al, 2020), the effectiveness of these
molecules in a real-world clinical setting has not been reported. Here
we report in a clinical setting the use of stem cell released molecules
combined with micro needling is a safe and efcacious means to
regrow hair in those patients with alopecia.
Case Report
Subjects
A total of twelve subjects were enrolled in our case stud ies.
Participants ranged in age. All subjects were recruited for the study and
treated by Dr. Michael Ryan, who is a practicing trichologist at the
Dubai Hair Clinic. Men and women eligible for inclusion in the trial
were in good general health with no evidence of systemic illnesses
(e.g., cardiac, psychiatric, or scalp disease). Patients known to be
hypersensitive to minoxidil were excluded, as were patients who
concomitantly used hair restorers or systemic drugs (steroids,
cy toto xic agen ts, va so dila t or s, ant i hy pert ens ive ag en ts,
anticonvulsant drugs, β-adrenergic receptor blockers, diuretics, or any
of the fol lowing spe cic ag ents: s pironolact one , ci met idine,
diazoxide, cyclosporine, or ketoconazole).
Procedure
This was a 12 - week trial conducted at 1 investigative site in the UAE.
The protocol and informed consent form were approved by the Dubai
Health Authority guidelines, and written informed consent was
obtained from each patient before enrollment in the trial. The test
solution with a dose of 1mL of assigned solution was applied to the
frontoparietal and vertex areas of the scalp for 18 weeks, daily both
morning and evening. In addition, on visits to the trichologist occurred
bi-weekly, at which time the test solution was applied and then the
scalp was microneedle. Microneedling was performed using an
Eclipse Micropen Elite, and the micro needling procedure included
multi directional passes (lateral, horizontal and 45 degrees to the
parting) of the device over the treated area at a depth of 0.75mm –
1.5mm, prior to application of the test solution. Variable depths of the
micropen were required in order to produce erythema at the target site,
some pin point bleeding was noticeable. Upon erythema being
noticeable, 5mLtest solution was then applied via a no-needle syringe
to the target area and the micro pen procedure was repeated with a
targeted approach. After the baseline visit (week 1), patients returned
to the clinic for efcacy and safety evaluations every 6 weeks through
week 12, then every 6 weeks through the end of the 18 -week trial.
Hair counts were obtained by using a Firey Pro 330T derma scope
using, a 1 cm2 target evaluation area in the thinning vertex scalp,
dened by an area consistently measured at 8cm and 20cm from the
center of the subject's eyebrows to the center of the vertex. Images
were captured at baseline and weeks 1, 6, 12, and 18, were calculated
by a computer program (TrichosciencePro V 1.7SE microscopic
evaluation software). This analysis was performed by a trained
trichologist. The resulting hair counts and hair thicknesses per square
centimeter were used to calculate mean change from baseline.
Product Ingredients
Human Stem Cell Conditioned Media, Human Fibroblast Conditioned
Media, Water, Glycerin, Larix Europaea Wood Extract, Camellia
Sinensis Leaf Extract, Santalum Acuminatum Fruit Extract, Citrus
Glauca Fruit Extract, Acacia Victoriae Fruit Extract, Trifolium
Prate ns e ( Cl over) Flow er Ex tract, Zinc Chlor id e, Gl ycine ,
Hydroxyethylcellulose, Dehydroacetic Acid, Benzyl Alcohol, Lactic
Acid, Sodium Hydroxide, Sodium Metabisulte.
Choice of Stem Cell Types for Deriving the Stem Cell Released
Molecules
Not all adult stem cells are alike. Elly Tanaka's lab (Kragl et al, 2009)
found that adult stem cells are tissue specic, meaning that adult stem
cells from a specic tissue are optimal for repairing and regenerating
that specic tissue. In these studies, we used adult stem cells to make
and release the molecules used in the test product that are derived from
the hair follicle and the skin surrounding the hair follicle. Cell types
used for therapeutic development must be chosen carefully for the sake
INTERNATIONAL JOURNAL OF SCIENTIFIC RESEARCH
Clinical Research
International Journal of Scientific Research
1
Volume - 11 | Issue - 04 | April - 2022 | PRINT ISSN No. 2277 - 8179 | DOI : 10.36106/ijsr
ABSTRACT
Introduction: Alopecia is a chronic dermatological disorder affecting millions of people, in which people lose some or all of the hair on their head.
Although alopecia has many forms, all are characterized as a chronic inammatory disease that affects the hair follicles. Alopecia often has
psychological consequences, including high levels of anxiety and depression.
Case presentation: We report hair regrowth in 12 patients with alopecia treated with adult stem cell released molecules in combination with micro
needling.
Conclusion: Adult stem cell released molecules in combination with micro needling is an efcacious, safe, and affordable treatment for alopecia.
KEYWORDS
Alopecia; Micro needling; Stem Cell Released Molecules; Hair Growth.
Volume - 11 | Issue - 04 | April - 2022
2
International Journal of Scientific Research
of safety and efcacy. Maguire (2019; 2021) and Conese et al (2020)
have reviewed why ADSCs are the preferred stem cell type for
developing therapeutics, especially for skin conditions. They review
why other stem cells types, notably bone marrow mesenchymal stem
cells (BMSCs), are a poor choice for many therapeutic applications.
For example, many lines of evidence from in vitro and in vivo studies,
including those in human BMSC transplants, nd that BMSCs and
their exosomes are carcinogenic and metastatic (Qi et a;, 2017; Luo et
al, 2020; Huang et al, 2020), owing to a number of factors, including
the fusion of BMSCs with cancer cells (Terada et al, 2002) or by the
education of BMSCs by cancer cells that yields a cancerous phenotype
in the BMSC (Nakata et al, 2017; Sai et al, 2019). Indeed, BMSCs
transplants in humans routinely increase the risk of cancer and death in
the recipient (Maguire, 2019B; Zhao et al, 2019). Further, exosomal
transfer of miRNAs from the bone marrow mesenchymal stem cells
may promote breast cancer cell dormancy, such that a state of breast
cancer may lay in wait for an appropriate signal (Ono et al, 2014).
These oncogenic factors have not been found in ADSCs or their
exosomes (Maguire, 2019), including the ADSCs and their exosomes
from cancer patients (García-Contreras et al, 2014).
RESULTS
Signicant hair growth was observed in all but one patients treated.
Although there were small changes to pigmentation color in some
candidate, pigment changes were unmeasurable in this study.
Terminal Hair Counts
In 11 of 13 patients signicant hair growth was achieved by the
combined use of microneedling and topical application of stem cell
released molecules (see Table 1).
Table 1. Individual counts of terminal hair units per given area in
the scalp of patients who were treated with a combination of micro
needling and stem cell released molecules. Values are presented for
before treatment and after treatment.
Figure 1. Example of hair count measurements, A. before treatment
and, B. after treatment. Green colored areas are computer
determined, and pseudo-color coded terminal hairs as distinguished
from vellus hairs.
Thickness (Diameter) of Individual Terminal Hairs
In 9 of 12 patients signicant hair thickness (diameter) was achieved
by the combined use of microneedling and topical application of stem
cell released molecules (see Table 2).
Table 2. Individual counts of terminal hair unit thickness (diameter)
in the scalp of patients who were treated with a combination of micro
needling and stem cell released molecules. Values are presented for
before treatment and after treatment.
Figure 2. Example photo used to measure hair unit thickness, A.
Before and B. After treatment.
DISCUSSION
The secretome derived from ADSCs and broblasts include PDGF,
FGFs, HGF, VEGF, IGF binding protein precursors, and bronectin (An
et al, 2021). PDGF has been found to induce and maintain the anagen
phase in the hair cycle in a mouse model, and HGF facilitates hair follicle
elongation (Lee et al, 2001; Yomita et al, 2006). VEGF increases hair
growth and size of the follicle by vascularization of the follicle (Yano et
al, 2001), and IGF-1 improves the migration, survival, and proliferation
of hair follicle cells (Gentile and Garcovich, 2019). FGFs secreted from
broblasts also promote hair growth by inducing the anagen phase in
resting hair follicles (Lin et al, 2015). Apoptosis is a normal process in
the hair cycle, and mesenchymal stem cells help to clear apoptotic cells,
following which they release PGE2 that decreases inammation (Zhang
et al, 2019). Thus, EVs from ADSCs, which have been involved in
apoptotic clearance, may contain PBE2 and reduce inammation in the
hair follicle, thus augmenting hair growth. Recent studies in a mouse
model of hair loss found that epithelial stem cells in the follicle escape
from their stem cell niche into the dermis in aged mice, thus contributing
to the miniaturization of the hair follicle (Zhang et al, 2021). Because
stem cells secrete factors that maintain the stem cell niche (Lee et al,
2016), a plausible partial explanation for how the secretome of ADSCs
facilitates hair growth is that the niche is maintained and inhibits stem cell
escape from the follicle. In general, during the cyclic quiescence and
activation of hair growth, hair follicle stem cells in the resting phase
constantly “sum” the input of activators and inhibitors, and when total
activators become dominant, the follicle enters growth phase (Lei and
Chuong, 2016). The molecules used in this study help to drive the sum of
signaling to that of activation.
In conclusion, we've provided evidence in a real-world, clinical setting
that a combination of micro needling and stem cell released molecules
(S2RM) induces a signicant increase in terminal hair counts and hair
thickness that is objectively measured using state of the art clinical
imaging, and that is subjectively observed by the clinician and patient
alike as signicant hair growth.
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PRINT ISSN No. 2277 - 8179 | DOI : 10.36106/ijsr
Subject
Sex
Age
Duration
Of Treatment
Weeks
Hair Density
Before (sq-cm)
Hair Density
After (sq-cm)
Ali
M
41
12
140
177
Nida
F
32
24
120
161
Aswain
M
39
12
107
140
Sal
M
45
12
102
248
Has
M
39
12
107
96
San
M
34
12
178
200
Jo
M
28
24
96
126
Sand
M
42
12
113
140
Nak
M
48
24
133
196
Son
M
49
24
42
70
Naz
F
44
12
81
111
Rav
F
62
24
117
285
Man
M
64
24
78
93
Ali
M
41
12
>30 µm): 87%
(64.2 per sq.cm)
>30 µm): 82% (75.3 per
sq.cm)
Nida
F
32
18
(>35 µm): 76%
(81.5 per sq.cm)
(>35 µm): 88% (68.5 per
sq.cm)
Aswain
M
39
12
(>30 µm): 85%
(85.2 per sq.cm)
(>30 µm): 79% (96.3 per
sq.cm)
Sal
M
45
12
(>30 µm): 82%
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sq.cm)
Has
M
39
12
(>30 µm): 85%
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(>30 µm): 92% (85.2 per
sq.cm)
San
M
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(107.4 per sq.cm)
(>30 µm): 86% (88.9 per
sq.cm)
Jo
M
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18
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(>35 µm): 84% (100.0
per sq.cm)
Sand
M
42
12
(>30 µm): 95%
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(>30 µm): 87% (79.3 per
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Nak
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48
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Son
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Naz
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per sq.cm)
Rav
F
62
18
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(25.9 per sq.cm)
(>35 µm): 71% (31.5 per
sq.cm)
Man
M
64
18
(>30 µm): 92%
(44.4 per sq.cm)
(>30 µm): 88% (77.8 per
sq.cm)
Subject
Sex
Age
Duration
Of Treatment
Weeks
Hair Thickness
Before (sq-cm)
Hair Thickness
After (sq-cm)
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