papilla proliferation = new hair follicles?

"researchers in Orlando have come a step closer to a natural treatment after successfully growing new hair using human stem cells.
The breakthrough was achieved after coaxing stem cells to become dermal papilla cells – a special type of cell which is vital to follicle formation.
Dermal papilla cells make up the top two layers of skin and they cause surrounding cells to form hair follicles. Hair loss occurs when papillae stop working."
http://www.dailymail.co.uk/sciencetech/article-2928737/A-cure-hair-loss-Scientists-grow-hair-rats-using-stem-cells-say-treatment-work-humans-too.html

Above was reported earlier this year. My question is this I have read over the years that there are several topical herbal extracts that induce dermal papilla proliferation eg Amla however these ointments have not been successful in creating new hair. Why then above are they claiming that coaxing stem cells to become dermal papilla would result in the genesis of new hair follicles. Also why would you need to grow these papilla outside the body and then inject them into the scalp when there are topical which are said to induce papilla proliferation in the scalp?

E. officinalis Linn and/or amla improves utilization of iron (metabolism) and helps with oxygenation via red blood
cells. It works by inhibiting 5-AR.

A lot of the research on "waking up" using stem cells using "technology." Although there are some internal stragties that help on some of the mechanisms discussed. Here's a look at some of the strategies:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4444612/

CausticSymmetry wrote:E. officinalis Linn and/or amla improves utilization of iron (metabolism) and helps with oxygenation via red blood
cells. It works by inhibiting 5-AR.

A lot of the research on "waking up" using stem cells using "technology." Although there are some internal stragties that help on some of the mechanisms discussed. Here's a look at some of the strategies:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4444612/

It also promotes DP proliferation, but several things do eg argan however if you can cause the DP to proliferate in the scalp by applying topical then why are they growing them outside and then injecting them into the scalp?

"Emblica (Phyllanthus emblica Linn.) Fruit Extract Promotes Proliferation in Dermal Papilla Cells of Human Hair Follicle
S. Luanpitpong, U. Nimmannit, V. Pongrakhananon and P. Chanvorachote

Abstract: Emblica (Phyllanthus emblica Linn.) has been used to promote the growth of hair in traditional medicine, however less is known regarding its pharmacological activities in hair follicles. To investigate the effects of emblica fruit extracts in terms of hair growth stimulation, the proliferative effects of extract in HaCaT keratinocytes and Dermal Papilla (DP) cells of human hair follicles were determined by MTT assay and cell counts. The results show that emblica extract stimulated proliferation of DP cells in a concentration-dependent manner, whereas it showed minimal effect on keratinocytes, suggesting that the extract might promote hair growth by prolonging the anagen phase through the proliferative effect on DP cells."

http://scialert.net/abstract/?doi=rjmp.2011.95.100

This is taken from the full study for the details on that:

Emblica (Phyllanthus emblica Linn.) 

Res. J. Med. Plant, 5 (1): 95-100, 2011

attributable to its high vitamin C and polyphenolic content (Scartezzini at al., 2006; Liu et al.,
2008). lmportantly, emblica fruits have been long used as a hair growth nourishment in traditional
Tibetan and Ayurvedic medicine (Dweck and Mitchell. 2005). Recent in vivo studies suggested
emblica as one of the herbs that acclaimed with hair growth promoting activity as it i composed
in the herbal formulations that effectively enlarge size and prolong the anagen phase of hair
follicles (Purwal at al., 2008; J adhav at al., 2009). However, its biological effects on follicular cells
are still largely unknown.

In the present study, we performed in vitro experiments using cultured human DP cells of
healthy hair follicles and HaCaT keratinocytes to investigate the effect of emblica fruit extract.

MATERIALS AND METHODS

Plant materials: Fresh emblica fruits were collected from Srakaew Province of Thailand, where
the climate is tropical. The samples were picked at commercial harvest time (May 2007) and selected
for the uniformity of shape and maturity.

Extraction and sample preparation: Fresh emblica fruits were cut into small pieces. The
materials (1 kg) were macerated and agitated in distilled water (5 L) and then filtered. The filtrate
was converted to powder form by spray drying. Yield of the extract was approximately 35%. The
extract contained 308-320 mg g“ of total phenoli calculated as Gallic Acid Equivalent (GAE)
using Folin-Ciocalteu’s reaction. Various samples of emblica extract were prepared by dissolving
the dry emblica extract powder in deionized water to the indicated concentrations (10-500 pg mL“).

Chemical reagents: Vitamin C (L-ascorbic acid) was obtained from Ajax Finechem (Auckland,
New Zealand). Ketoconazole was obtained from Crosschem Intern ational Company, Derb and Co.
(Lugano, Switzerland). 3-(-1,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was
obtained from Sigma Chemical, Inc. (St. Louis, MO, USA).

Cells and cell culture: Human follicle dermal papilla cells (DP cells) were obtained from
PromoCe1l (Heidelberg, Germany). Cells were cultured in DP cell growth medium (PromoCell)
containing 100 units mL" of penicillin and 100 pg mL" of streptomycin (Gibco, Gaithersburg, MA,
USA) in a 5% CO, environment at 37°C.

Human immortalized keratinocyte HaCaT cells were obtained from Cell Lines Service
(Heidelberg, Germany) and maintained in Dulbecco’s modified Eagle's medium (DMEM; Gibco,
Gaithersburg, MA, USA) supplemented with 2 mmol L" L-glutamine, 10% fetal bovine serum,
100 units mL" of penicillin and 100 pg mL“ of treptomycin in a 5% CO, at 37°C.

Cell proliferation: Cell proliferation was determined by MTT colorimetric assay (Mosmann, 1983)
and cell counts. For MTT asay, cells at a density of 5X10’ cells per well (3 replicate per treatment)
were seeded in a 96-well plate and incubated with emblica fruit extracts (0-500 pg mL") for 24 h.
Alter that cells were incubated with 500 pg mL" of MTT for 4 h at 37°C. The upematant was then
removed and dimethylsulfoxide (DMSO) was added to dissolve the formazan product. The intensity
was spectrophotometrically measured at 550 nm using an ELISA reader (Anthros, Durham, NC,
USA). All analyses were performed in three independent replicate cultures. The intensity readings
were normalized to a non-treated control and represented in the terms of relative cell proliferation.

For cell counts, cells at a density of 2X10‘ cells well“ were plated in 24—well plates. Cells were
cultured in a control medium or emblica extract-containing medium. At each time point (0, 24 and
48 h), the number of DP cells was counted using hemocytometer under light microscope.

Statistical analysis: Statistical analysis is performed using the Student's t-test. A p-value of less
than 0.05 would be considered as statistically significant.

RESULTS

Effects of emblica fruit extract on cell proliferation: The cell growth promoting effect of
emblica fruit extrad. was investigated in immortalized keratinocytes (HaCaT) and primary human
hair DP cells using MTT assay. Cells were incubated with various concentrations (10-500 pg mL")
of emblica enract and cell proliferation was determined after 24 h. The results indicated that
emblica extract caused a dose-dependent increase in DP cell proliferation (l.ll:h0.l8-fold,
1.l9:|:0.19-fold, 1.88:E0.05-fold, 2.31:H).l l-fold and 2.67:|'.-0.39-fold at 10, 50, 100, 250 and

500 pg mL" of extract, respectively). whereas. it had minimal proliferative effect on HaCaT
keratinocytes (Fig. Is). At various concentrations ranging from 100-500 pg mL", the extract
significantly increased DP cell growth with the maximal effect by approximately 3-fold compared
to the non-treated control. Notably, at the treatment dose of 500 pg mL", the proliferative effect
of emblica extract was only slightly increased compared to that of 250 ug mL".

To confirm the proliferative effect of emblica extract on DP cells, DP cell numbers were counted

after cultivation in emblica extract (100 pg mL“)-containing medium at various times. Consistent
with the MTT assay, the number of DP cells markedly increased in the emblica-treated group over

a non-treated control (Fig. lb).

Fig. 1: Emblica fruit extract increases DP cell proliferation. (a) I-IaCaT and DP cells were treated
with various concentrations of emblica extract (10-500 pg mL") for 24 h and cell
proliferation was determined by MTT assay and (b) DP cells were cultured in emblica
extract (100 pg mL")-containing medium. Cell numbers were counted using
hemocytometer at 24 and 48 h. Values are mean of triplicate samplesi=SD. *p<0.05 versus
non-treated control


Fig. 2: Proliferative effects of emblica extract, vitamin C and ketoconazole on DP cells. Cells
were treated with emblica extract. (100 pg mL“), vitamin C (50 pg mL") or ketoconazole
(50 pg mL“) for 24 h and cell pmlifesration was determined by MTT assay. Values are mean
of triplicate samples=|:SD. *p<0.05 versus non -treated control

Proliferative effecis of emblica fruit extract and hair growth stimulators on DP cells:
Previous study has shown that vitamin C derivative can induce hair gowth in vivo and in vitro.
This action of vitamin C derivative was hown to be caused by its bility to induce DP cell growth,
while having a minimal effect on other follicular cells (Sung et al., 2006). Moreover, clinical
evidence indicated that anti-fungal ketoconazole possesses an ability to promote hair growth
(Pierard-Franchimont at at, 1998; Ellis nd Sinclair, 2008). Therefore, we compared the
proliferative effect of emblica extract with vitamin C and katoconazole.

Figure 2 shows that emblica extract, vitamin C (L-ascorbic acid) and ketoconazole exhibited
significant proliferative effects on DP cells. The proliferate effects of emblica extract at its minimal
effective concentration (100 pg mL“) was stronger than those of vitamin C and ketoconazole at the
concentration of 50 pg mL" (2.03:h0.21-fold vs. 1.25:t0.l0-fold and 1.15eh0.09-fold, respectively)
(Fig. 2). Notably, higher concentrations (>50 pg mL") of vitamin C and ketoconazole could not be
used in the present study due to their cytotoxic effect.

Now that my inflammation has been totally eradicated, and hairs are thickening elsewhere, I'm hoping that matrix cells in my bald areas will begin to proliferate. If this doesn't happen sometime soon, then I am pretty much fucked because I don't know how to trigger these growth factors in said cells. Yet, the reason why I am optimistic, is down to the fact that alopecia areata sufferers regrow hair after many years of being bald. AA and MPB share a similar pathophysiology in regards to inflammation destroying DP matrix cells, therefore if healing and regrowth can take place in AA sufferers, then the same principle should apply to MPB. It just depends how long this will take to happen (if it does).

Additionally, we know that the follicles have not been destroyed, they are just in the resting phase, hence micro-vellus hairs being visible all over the bald areas under a certain angle of light. This suggests to me, that the only component - required for terminal hair growth - is these DP matrix cells.

Perhaps something like this:

Facial Plast Surg. 2014 Apr;30(2):219-24. doi: 10.1055/s-0034-1371896. Epub 2014 May 8.
Platelet-rich fibrin matrix (PRFM) for androgenetic alopecia.

The objective of this study was to determine the effect of platelet-rich fibrin matrix (PRFM) treatment on androgenetic alopecia. Prospective cohort study of 15 (9 male and 6 female) subjects with androgenetic alopecia for at least 1 year who were treated with intradermal injections of autologous PRFM three times on a monthly basis. Hair density indices were measured in triplicate in the same area of the scalp before the treatment and 1, 2, 3 and 6 months after initial treatment. Hair density index (HDI) measurements were obtained and compared with pretreatment values for each subject. After a series of three intradermal PRFM injections, hair density indices increased significantly at 2 (47.4 ± 22.7%, p = 0.0031) and 3 (106.4 ± 56.9%, p = 0.0277, paired t-test) months after the initial treatment, and approached statistical significance at 6 months (75.1 ± 46.82%, p = 0.0606) after the initial treatment. Patients who achieved greater than 25% increase in HDI by 2 months after the initial treatment were more likely to have greater than 25% improvement at 6 months after the initial treatment (100 vs. 16.7%, p = 0.0476). Androgenetic alopecia affects a significant number of both men and women. A series of intradermal injections of autologous PRFM increased the HDI in patients with androgenetic alopecia at 2 and 3 months after initial treatment; this improvement approached statistical significance at 6 months after initiating treatment. Autologous PRFM injections may be a valuable treatment for androgenetic alopecia, particularly in cases with mild hair loss. The level of evidence is level 2.

Xenon wrote: the reason why I am optimistic, is down to the fact that alopecia areata sufferers regrow hair after many years of being bald. AA and MPB share a similar pathophysiology in regards to inflammation destroying DP matrix cells, therefore if healing and regrowth can take place in AA sufferers, then the same principle should apply to MPB.

MPB is called scaring alopecia because micro fibrosis (scaring) results in a shiny scalp, not so with AA. Once a hair follicle has been dormant for more then 2.5 years it dies and it cant be resurrected so best you can do is return your hair to how it was 3 years ago.

They have also experimented with injecting a cocktail of blood plasma mixed with herbal extracts known to increase DP proliferation and have reported successful results. Perhaps these herb extracts cant penetrate the scalp in which case a dermal roller is going to be needed.

H4ir wrote:

Xenon wrote: the reason why I am optimistic, is down to the fact that alopecia areata sufferers regrow hair after many years of being bald. AA and MPB share a similar pathophysiology in regards to inflammation destroying DP matrix cells, therefore if healing and regrowth can take place in AA sufferers, then the same principle should apply to MPB.

MPB is called scaring alopecia because micro fibrosis (scaring) results in a shiny scalp, not so with AA. Once a hair follicle has been dormant for more then 2.5 years it dies and it cant be resurrected so best you can do is return your hair to how it was 3 years ago.

From my knowledge and observations:

1. The shiny parts of my scalp are shiny due to increased sebum production. When I wipe this skin with a tissue it becomes matt and dry (normal skin texture).

2. AA sufferer's DP cells experience auto-immune attack in a similar way MPB sufferers do; the end result is inflammation and miniaturization of the follicle. These cells are constantly renewed so long as the hair bulge is intact and signaling pathways are active. I read a study in which the papilla itself was surgically removed, but new follicles were formed because the bulge was left in place.

3. The bulge is also responsible for growth and maintenance of the sebaceous / sweat glands. The fact that we produce so much sebum and sweat from the balding areas tells us that the hair bulge is intact and has not been replaced with scar tissue.

4. Under a certain angle of light I can see what was my hairline, now in a vellus state. This means that the papilla is still existent and still producing hairs, and has not been replaced with fibrotic scar tissue - it is just in the telogen phase.

So, if the papilla and hair bulge still exists, then there is hope that the follicle can re-enter the anagen phase.

ETA: my uncle's girlfriend has suffered alopecia totalis throughout her life. She was totally bald for a decade and then her hair started to regrow. Therefore, this is further evidence that healing and regrowth can take place, despite cells being subject to auto-immune attack.

The scalp on the vertex develops micro scaring which causes shininess and tight skin in the same way as if you got cut by a knife the skin around the cut will be shiny and tight in comparison to normal skin. When a scar develops around a hair follicle it chokes it eventually burrying it in the skin, once that happens its dead. The pore, hole the hair follicle would normally grow out of is closed off by the scar tissue. If you were to shave your head and examine with a magnifying glass the non balding areas with the balding areas you will see less pores/holes where hairs grow out of because they have been closed off.

The follicle isn't dead. The papilla is still producing vellus hairs. So how can it be dead? I can see every single hair in micro-vellus form. This means it is merely in the telogen phase.

If the pore hole was closed off by scar tissue, then the bald areas would produce no sebum. You know why? Because the sebaceous gland is located inside the follicle. This means that the follicle must be open if sebum is being secreted onto the scalp surface. Sebum must pass through the follicle in order for it to reach the scalp surface.

The scalp is shinier than elsewhere, not because of scar tissue, but because a) increased sebum production b) the tissue is pulled a little tighter across the cranium than elsewhere.

"Dermal fibrosis in male pattern hair loss: a suggestive implication of mast cells.
Won CH1, Kwon OS, Kim YK, Kang YJ, Kim BJ, Choi CW, Eun HC, Cho KH.
Author information
Abstract

A relationship has been suggested between mast cells (MCs) and male pattern hair loss (MPHL), because of histological evidence of perifollicular fibrosis and increased mast cell numbers. Two paired punch biopsies were taken from balding vertexes and non-balding occipital promontory areas of ten patients with MPHL (Ludwig-Hamilton IIIv to IV) and from five normal subjects aged from 20 to 35 years. Masson trichrome and Victoria blue staining were performed to observe collagen frameworks and elastic fiber structures. Numbers of immunoreactive MCs stained with anti-tryptase or anti-chymase antibody were counted. It was found that collagen bundles were significantly increased in balding vertexes than in non-balding occiput scalp skin. A near 4-fold increase in elastic fibers was observed in both vertex and occiput scalp skins with MPHL versus controls. Total numbers of MCs (tryptase-positive) in site-matched scalp samples were about 2-fold higher in MPHL subjects than in normal controls. Percentage elastic fiber (%) was found to be relatively well-correlated with tryptase and chymase-positive MCs. These findings suggest that accumulated MCs might be responsible for increased elastic fiber synthesis in MPHL, and indicate that future investigations are warranted."


" Konstantinova N, Korotkii N.G, Sharova N, Barhunova E, Gaevski D. Nioxin Research Inc, AtlantaIrreversibility of hair follicle changes after 30 months of Androgenetic Alopecia.
, USA Moscow Medical University

We studied horizontal and vertical biopsy from 15 caucasian 24-41 year old males diagnosed with bitemporal recession Androgenetic Alopecia (AA) for 1.5 –18 years (average 7.4 years). All 15 biopsies were stained with H&E, Van Gieson and with other collagen specific stainings. 1. Eleven pts with AA longer than 3 years had perifollicular fibrosis - collagen fibers were compact and formed a small scar-like formation around each anagen hair follicle(HF). Two patients - 33 year old with 18 month AA and 23 year old with 20 month AA did not have these hair follicle changes. Two 26-year-old patients with 30 and 36 month AA respectively were found to have some not so severe collagen fiber changes. 2. Infundibulum of HF dilatated 124-192 mm and most of them covered with keratinazed plug lacking normal hair shaft growth. 3. Decreased number of hair follicles 1.75-2,45 per sq. mm from 3.5-5 per sq. mm in control group. 4. None of anagen HF was situated in subcutaneous fat. We showed a correlation between length of the AA and severity/ thickness of perifollicular fibrosis. The result of this study is that any treatment of AA is recommended to start earlier than 30 months from first signs of AA. This should prevent irreversible collagen changes associated with “fibrotic incapsulation” of most anagen HF in involved areas, which usually leads to loss of normal blood supply, innervation, and subsequent miniaturization and prevention of hair from normal cycling."

I've read studies on perifollicular fibrosis before, but I've also read various MPB studies which state that the reason why there is no matrix cell proliferation is due to lack of CD34+ progenitor cells within the bulge. So what's the truth here? I'm inclined to believe in the latter based upon the aforementioned reasons: 1. Alopecia areata sufferers are known to regrow their hair after many years of follicular inflammation 2. the papilla is growing vellus hairs through a pore opening. 3. Sebum / sweat is being produced, meaning the hair bulge is still functioning.

However, I don't know everything and don't want to be dismissive of the fibrosis factor. I'm just going by the information I have gathered over the years. As mentioned, my inflammation has been entirely eliminated, so we'll see if hair starts to grow back soon enough. Maybe, maybe not. If fibrosis really is the issue here, then perhaps we are fucked after all, and all of this study is a waste of time.

"and most of them covered with keratinazed plug lacking normal hair shaft growth"

Well I guess the rate at which this fibrosis occurs varies between individuals and diet must play a major role. Sugar is really bad !

With hair loss people need to act asap with treatment to prevent this fibrosis because its like trying to sprout seeds on hardened ground once it is excessive. The reason the hair diminishes in the first place is the fibrosis restricts the micro capillaries around the hair bulb so it cant receive nutrients. As the hair shaft starts to miniaturise in diameter the pore from which it grows also starts to decrease in diameter with fibrosis of its edges it is inflexible. Last stage is the pore is closed off with a "kertinazed plug" and then we are fucked!

To avoid this as much as possible we need to:

Exfoliate the scalp by brushing and perhaps using an occasional chemical peel.

Avoid sugar and high carb diet.

Load our bodys with anti inflammatory including supplements

Boost the blood circulation to our scalp, red light, Vasodilators, massage, brushing.

Reduce androgens in scalp, topical dht inhibitor