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Remedy Publications LLC.
Annals of Stem Cell Research & Therapy
2021 | Volume 5 | Issue 1 | Article 1041
1
Mesenchymal Stem Cell Conditioned Media Induces Hair
Regeneration in Alopecia Areata: A Case Study
OPEN ACCESS
*Correspondence:
Anand S Srivastava, Department of
Stem Cell Therapy, Global Institute
of Stem Cell Therapy and Research,
4460 La Jolla Village Drive, San Diego,
92122 CA, USA, Tel: +1 (858) 344-
2492;
E-mail: anand@giostar.com
Received Date: 24 Feb 2020
Accepted Date: 25 Mar 2021
Published Date: 29 Mar 2021
Citation:
Mahmood A, Seetharaman R, Kshatriya
P, Patel D, Mishra R, Srivastava AS.
Mesenchymal Stem Cell Conditioned
Media Induces Hair Regeneration in
Alopecia Areata: A Case Study. Ann
Stem Cell Res Ther. 2021; 5(1): 1041.
Copyright © 2021 Anand S
Srivastava. This is an open access
article distributed under the Creative
Commons Attribution License, which
permits unrestricted use, distribution,
and reproduction in any medium,
provided the original work is properly
cited.
Case Report
Published: 29 Mar, 2021
Introduction
Alopecia is characterized by hair loss in one or more regions of scalp or body. e prevalence
rate is 2.1% in USA with global incidence risk rate of 2%. Several factors including stress, hereditary,
nutritional disorders, and thyroid functional abnormalities, psychological, genetic, and immune
disorders have been held responsible for triggering of disease. ough, the disease is not life
threatening, it is associated with aesthetic values and self-dignity. e commencement of disease in
early age can cause psychological concerns like low self-esteem, depression, or anxiety, aecting the
quality of life. Alopecia Areata (AA), specically, is an autoimmune disorder caused due to targeting
of immune-privileged Hair Follicles (HF) by T cells. AA is characterized by patchy scalp hair loss
which may progress to complete scalp hair loss (alopecia totalis) or complete body hair loss (alopecia
universalis). e incidence of AA is associated with co-occurrence of other autoimmune disorders
like psoriasis, vitiligo, systemic lupus erythematosus, rheumatoid arthritis, and inammatory
bowel disease. Further, thyroid related abnormalities are frequently found in AA patients. Diabetes
mellitus co-occurrence with AA has been reported in patients with increased insulin resistance
[1]. e current available interventions include topical minoxidil, topical anthralin, intralesional
corticosteroids injections; topical corticosteroids contact sensitizers, and Psoralen plus Ultraviolet
an Irradiation (PUVA) therapy. ese therapeutic approaches are either partially successful or are
associated with signicant side eects. HF transplant is mediated by advanced surgical procedures
involving implantation of hair follicles to region without hair. e success of procedure is limited
due to poor availability of donors, low survival of transplants, painful procedures, and expensive
treatment. Further, the results achieved are oen temporary [2].
Strategies based on stem cells are under investigation for their ability to regenerate the lost
hair follicle and stimulate hair growth. Mesenchymal Stem Cells (MSCs) particularly have drawn
attention of researchers and clinicians owing to ease of availability, immune-privileged nature and
regenerative capacity. MSCs are multipotent cells present in multiple tissues including adipose,
umbilical cord, bone marrow, hair follicle, dental pulp and liver. ese are characterized by
high proliferative capacity, plastic adherence, and spindle shape morphology. Some studies have
demonstrated the therapeutic ecacy MSCs in HF regeneration [3,4]. However, the regenerative
potential of MSCs has been attributed to the growth factors they secrete. It has been reported that up
Abstract
Alopecia Areata (AA), a hair loss condition caused primarily by autoimmune disorder, aects
signicant population globally. e disease hampers quality of life causing anxiety and low self-
esteem in patients. is case reports discuss about condition of an alopecia areata patient and her
subsequent treatment with Mesenchymal Stem Cell derived Conditioned Media (MSC-CM). e
severity of disease was assessed by calculating Severity of Alopecia Tool (SALT) score which was
6%. e topical treatment for two months resulted in visible hair growth in vertex scalp area with
SALT score approaching 0%. MSCs secrete number of growth factors, cytokines, lipid vesicles,
and paracrine molecules in surrounding media which may regulate growth of hair follicles,
vascularization, and morphogenesis. is report indicates that topical treatment of MSC-CM
can alleviate alopecia condition and promote hair growth. is may be mediated through passive
diusion of paracrine factors and exosomes.
Keywords: Mesenchymal stem cells; Conditioned media; Aopecia areata; Hair regeneration;
Hair regrowth
Anjum Mahmood1, Rajasekar Seetharaman1, Prashant Kshatriya1, Divyang Patel1, Rangnath
Mishra2 and Anand S Srivastava2*
1Department of Stem Cell Therapy, Global Institute of Stem Cell Therapy and Research Pvt. Ltd, India
2Department of Stem Cell Therapy, Global Institute of Stem Cell Therapy and Research, USA
Anand S Srivastava, et al., Annals of Stem Cell Research & Therapy
Remedy Publications LLC. 2021 | Volume 5 | Issue 1 | Article 1041
2
to 80% of stem cell mediated regeneration is contributed by secretory
factors of their secretome [5]. In this report, we have investigated the
therapeutic eect of MSC derived Conditioned Media (CM), in a
patient with alopecia areata, on observational basis.
Case Presentation
Patient
A 45-year-old female patient suering from alopecia areata for 5
months approached our center. e preliminary examination of scalp
region showed presence of small bare non scarring patch in vertex
area. e severity of disease was assessed by calculating Severity of
Alopecia Tool (SALT) score. e SALT score was assessed to be 6%.
e SALT score calculation involves the percentage of hair loss in
eected area in particular scalp region [6]. e patient was under
mild anxiety and nervousness due to the persistent disease.
Preparation of MSC-conditioned media
Before procedure informed consent of patient was collected. e
detailed procedure has been discussed previously [7]. Briey, 100
ml of adipose tissue was collected from the patient by lipoaspiration
from waist area by a trained plastic surgeon under the aseptic
environment. e cells were processed in an ISO 9001: 2015 certied
and DSIR (Department of Scientic and Industrial Research)
recognized laboratory. e lipoaspirate was processed in a biosafety
laminar airow chamber. MSCs were isolated from adipose tissue by
mechanical agitation including vertexing. Aer centrifugation, the
resulting pellet was cultured in DMEM medium (Invitrogen, Paisley
UK). Supplemented with 10% fetal bovine serum (FBS) and 1%
penicillin/streptomycin, at 37ºC in humidied atmosphere containing
5% CO2. e media were changed aer every 3 days. Approximately
5 × 106 MSCs of passage 1 were seeded in each T175 culture ask
(n=5) containing 30 ml of DMEM medium supplemented with
10% FBS. MSCs were analyzed for spindle shaped morphology and
any contamination using a phase-contrast microscope. When cells
reached 70% conuence at passage 2, the culture media was replaced
with serum-free DMEM. Post 72 h of incubation, the culture media
was separated and collected. e culture media was then centrifuged
at 2000 rpm for 5 min to remove the cell debris, ltered through 0.22
m lter, and then concentrated by ultra ltration using centrifugal
ltering units with a cut-o value of 3 kDa (Amicon Ultra-15;
Millipore, MA), according to the manufacturer’s instructions. e
concentrated 30% MSC-CM in cream base was prepared for topical
application. e cream was applied on aected area of scalp once
daily for a period of two months. e patient was monitored for
development of any adverse events or severity of disease. During this
period, patient did not receive other treatment for alopecia.
MSC-CM reinstates hair growth
e patient was evaluated at 1 month and 3 months post 1st
application of MSC-CM (Figure 1). No adverse events associated
with MSC-CM application were recorded. Before treatment single
bare hairless oval region was observed at vertex of scalp. At 1month
follow-up, the hair growth was observed with marked reduction in
circumference of hairless patch. Post 3 months, signicant dierence
was detected with visible increased hair count and density in eected
scalp region. e SALT score reduced to approximately 0% with
continued improvement and follow up to 6 months. Aer hair
growth restored, patient was out of alopecia related anxiety, improved
condence level and led enhanced quality of life.
Discussion
Alopecia areata is marked by patchy hair loss caused primarily
due to autoimmune disorder. e overactive immune cells abolish
hair follicles causing non-scarred hair loss. Dermal papilla cells are
the key regulator of initiation and regulation of hair growth. ey
control surrounding niche and mediate cross-talks with stem cells.
Hair growth is primarily regulated by Erk activation and Wnt
signaling. e stimulation from dermal papilla cells, activate hair
follicles through activation of Wnt-β catenin pathway [8]. In patchy
Alopecia, though progenitor cells are lost, hair follicle stem cells are
preserved. is kind of Alopecia can be reversed through application
of multipotent stem cells [9]. MSCs are the most investigated
multipotent stem cells, owing to immune-privileged character, for
their role in tissue regeneration and wound healing. MSCs promote
angiogenesis, inhibit cell apoptosis, immunomodulate host immune
system and demonstrate antibrotic activity. e functional roles of
MSCs are facilitated through secretory cytokines and growth factors.
In the case discussed here, adipose derived MSCs were used to generate
conditioned media (MSC-CM) comprising of secretory molecules.
MSC-CM consists of cell free system enriched with growth factors,
chemokines, cytokines and exosomes. e interventions subside
requirement of immune-compatibility matching and exclude the
possibility of teratoma formation. Moreover, the treatment procedure
involved topical application of MSC-CM mixed with cream base.
e patient had received several treatments, without any noticeable
change, before considering MSC-CM based intervention. Signicant
Figure 1: Effect of MSC-CM on Alopecia areata. (a) Vertex area of scalp of patient showing hairless patch; (b) Partial recovery of hair on scalp post one month of
follow-up; (c) Nearly complete covering of hairless patch with hairs post three months of follow-up of treatment.
Anand S Srivastava, et al., Annals of Stem Cell Research & Therapy
Remedy Publications LLC. 2021 | Volume 5 | Issue 1 | Article 1041
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improvement was observed in hair growth, aer one month of MSC-
CM topical application. e size of hairless patch was reduced, with
visible increase in hair count. ree months of treatment resulted in
eective resolution of alopecia. e SALT score reduced to 0% from
initial 6%. e notable recovery was continued for 6 months follow up
duration. Based on previous reports, it can be hypothesized that scalp
hair augmentation might have mediated through paracrine factors
secreted in MSC-CM. Vascular Endothelial Growth Factor (VEGF)
reported in MSC-CM, augments hair regeneration by promoting
perifollicular vascularization. Similarly, Hepatocyte Growth Factor
(HGF) induces follicular growth by stimulating β catenin expression.
e secretion of Insulin like Growth Factor -1 (IGF-1), has been co-
related with migration, survival, proliferation, tissue remodeling and
growth cycle of HF. IL-6 and M-CSF were reported to mediate hair
growth in wounds. Another key secretory protein, Platelet-Derived
Growth Factor (PDGF), trigger growth by promoting hair cycle to
enter anagen phase [10,11]. Overall, growth factors through complex
mechanism, in interdependent manner, induce proliferation of
dermal papillae cells by modulating cell cycle via up regulation of
cyclin D1 and CDK2. e factors activate Erk and Akt signaling
pathway [12].
ough, pre-clinical and clinical studies have investigated eect
of MSC-CM on hair growth and demonstrated its positive eect;
mainly intradermal injections were used, which is invasive in nature
[13]. Shin et al. used micro needle roller prior to topical application
of conditioned medium of adipose tissue derived stem cells [14].
Frequent usage of intradermal injections is challenging for patients
as it requires regular clinic visits, cause pricking discomfort, and
impose fear of infection. is report highlights simple approach of
topical application of MSC-CM for alopecia patients in self applicable
mode without distress and possibility of infection. e microltration
procedure used to lter MSC-CM make peptides size even, remove
aggregates and improves diusibility across the scalp [15]. Further,
MSC-CM contains lipid bound exosomes which are enriched in
growth related proteins, DNA and RNA. ese exosomes can integrate
with lipid membrane of hair scalp cells and facilitate the transfer of
liposomal materials. e exosomal content play signicant role on
regulating key signal pathways aecting hair growth. Exosomes
carry Wnt proteins on their surface which can induce activation of
β-catenin signaling pathway involved in hair growth [16,17].
Conclusion
e report discussed here, demonstrate induction of hair
growth in alopecia patient through topical treatment of MSC-
CM. ough, experimental investigations have indicated towards
success of intradermal applications of MSC-CM, inadequate data
is available to provide conclusive evidence on topical application.
Route of administration signicantly inuences ecacy of biological
molecules. is report provides an early clue on possible passive
diusion of signaling molecules through scalp. Detailed investigation
involving randomized placebo control studies are required to provide
insight on ecacy and tolerability of treatment. Besides, other
variables including source of MSCs, culture conditions (hypoxic or
normal/3D or 2D culture) and formulations, can potentially alters the
ecacy and require detailed analysis.
Funding
is study is funded by the Global Institute of Stem Cell erapy
and Research, San Diego, California, USA.
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