Reversal of hair loss in Mice with “sugar gel”

[Mauritio]

Glutamic acid.
Not exactly glutamine, but related .

Has very impressive hair boosting effects in this study, especially the 1% and 10% solution .

https://www.nature.com/articles/s41598-021-94816-y/figures/3

@CrumblingCookie I haven't fully read it, but I feel like this study explains a lot of what we're wrestling with.

It seems that glutamine drives hair follicle growth from stem cells (HFSC) to outer root shaft cells which support the growing hair in the anagen phase. Interestingly this process is triggered by glutamine inhibiting aerobic glycolisis and boosting TCA cycle. So generally this could be achieved by substances that supress glycolisis and boost TCA cycle.

On the other hand they show that excessive glutamine or blocking mtorc2, long term depletes the HFSC pool, and I guess over long time frame that would lead to impaired regeneration and thinning hair.

This is also explains why they saw such drastic effects in the study above, but I'm not sure if that would be the same long term.

So, maybe cyclic addition of glutamine would be best. Between 1-10% , maybe 1 month on 2 months off or sth like that ?

https://pubmed.ncbi.nlm.nih.gov/32905798/

[CrumblingCookie]

@Mauritio said:

I discussed the hair metabolism in this post
https://bioenergetic.forum/post/41937

Sorry I had missed this post. It's essentially the same study and authors we both dug out there.

@Mauritio said:

On the other hand they show that excessive glutamine or blocking mtorc2, long term depletes the HFSC pool, and I guess over long time frame that would lead to impaired regeneration and thinning hair.

This is also explains why they saw such drastic effects in the study above, but I'm not sure if that would be the same long term.

So, maybe cyclic addition of glutamine would be best. Between 1-10% , maybe 1 month on 2 months off or sth like that ?

The "spatiotemporal" effects really open up a different angle to determine suitable combinations and timings of different topical substrates. We wouldn't want a limited phase of fully engaged "hair growth bloom" followed up by depletion of the underlying stem cells and exacerbated balding.

It's very important that when I think of true hair loss reversal I am thinking of reactivated dermal papilla. Whereas feeding preexisting hair follicles only impacts hair thickness and rate of growth.
Of course outer sheath and hair thickness and follicle formation are crucial for robustness and rootedness of the hair follicle and for distinguishing proper head hair from vellus hair.
So we do need stimulation of both. Maybe in cyclic variation of topical regiments.

I've discovered a metabolic study in which the authors set up a 3D sphere culture to generate hair follicles from dermal papilla (DP) activating skin epithelial stem cells:
Glucose metabolism regulates expression of hair-inductive genes of dermal papilla spheres via histone acetylation

• "Here we show that active DP exhibit robust aerobic glycolysis. We observed decrease of signature genes associated with hair induction by DP in presence of low glucose (2 mM) and glycolysis inhibitors. Moreover, hair shaft elongation was attenuated by glycolysis inhibitors. Interestingly, excessive glucose is able to increase the expression of hair inductive genes and elongation of hair shaft. We also observed glycolysis-mediated histone acetylation is increased and chemical inhibition of acetyltransferase reduces expression of the signature genes associated with hair induction in active DP. "

• "energy metabolism has been shown to be one of the major cues during hair follicle formation. Especially, lactate dehydrogenase (LDHA), an enzyme involved in anaerobic glycolysis is required for hair follicular stem cell activation."

• "To determine whether sDP exhibit anaerobic or aerobic glycolysis, we measured pyruvate and lactate levels. sDP showed higher pyruvate levels but not lactate levels, which indicating aerobic glycolysis is activated by sDP (Fig. 1e,f)."

-->DP also rely on aerobic glycolysis. So no obvious contradictions between HF and DP so far.
Yet I am wondering what happens with all that excess pyruvate in HF and DP:
@Mauritio said:

...Aerobic glycolisis leads to copious amounts of intermediate generation like lactate and pyruvate . Which can be used to build nucleotides and keratin for new hair . In OxPhos the carbon goes towards CO2.

• "In addition to ATP production, SCs utilize glucose-derived carbons for lipid synthesis and for production of hexosamines, ribose, glycerol, serine, and glycine." *

Anyway, (excessive, 10-20mM) glucose concentrations (/extra energy) go hand-in-hand with histone acetylation and epigenetic activation of (dormant) DP under assumed normal circumstances:

• "Expression of DP signature genes including BMP4, LEF1, WNT5A, VCAN and FGF7 were decreased from sDP in presence of low glucose (2 mM) compared with physiological of glucose (5.5 mM) without alteration in cellular viability (Fig. 2a–f)."

• "Interestingly, excessive glucose (10 mM) supplement increased expression of the signature genes associated with hair induction (Fig. 2p-t). [...] We also found that hair shaft elongation was enhanced by increased glucose concentration (Fig. 2w,x).

Which seems to underscore the first and foremost importance of energy substrates but also makes me wonder under what (sick or altered) conditions and toeholds and auxilliary cells non-energy-subtrate HDACi or DNMTi or HMTi or LSD/KDM and coriander and apple phenols etc exert their action in this context.
If the extent of glycolysis directly drives histone acetylation in DP and HFs then more isolated benefits can be expected by reducing histone or DNA methylation back to prior physiologically states. Topical EGCG, decursin, apigenin?
The reducing qualities of plant phenols may be necessary whenever increasing the small proportion of OXPHOS in HF and DP, e.g. by NMN? Or generally for the larger proportions of OXPHOS in the afferent capillaries?

Interestingly, feeding ketones/fatty acids to hair follicles inhibits the PPP synthesis of ribonucleotides (your 1991 HF study. Don't know what the actual effects of this will be):

• "Both palmitate and beta-hydroxybutyrate inhibited glucose flux through the pentose phosphate pathway. "

@Mauritio said:

In wondering what other more benign amino acids do for the hair, like glycine ?

• "The suggestion that glutamine may be an important metabolic substrate in skin is strengthened by one further observation: the concentration of most amino acids in sweat reflect, largely, their concentration in blood; but the concentration of glutamine in sweat is only 3% of that of blood, 18"

which is taken from a 1974 study on healthy subjects during sauna bathing, i.e. an increase of metabolism surely met by increased tissue perfusion yet probably not by sufficiently increased glucose utilization. Are excessive sauna-goers and endurance sportsmen balding more widely and quickly?

Leucine and (sources of) sulfate may be worth a look because of biglycan as a significant messenger to Wnt activation for hair follicle renewal:
Dermal papilla cell-secreted biglycan regulates hair follicle phase transit and regeneration by activating Wnt/β-catenin
Activated VDR is an important factor for Wnt/β-catenin activation. So any of the numerous pathogens which express antagonizing ligands will suppress Wnt/β-catenin, as would excessive D3 and 25-OH-D3.

For dermal papilla functions, activating the Akt pathway and increasing the
level of Bcl-2 as well as VEGF and IGF-1 all seem important. DP also excrete powerful exosomes which induce their own activation via Wnt/β-catenin in neighbouring tissues. Reminds me quorum sensing in bacterial cultures. The more adjacent hair you have, the more hair you keep/get:

• "A growth factor cocktail of keratinocyte-conditioned media such as VEGF and IGF-1 was found to significantly increase the proliferation of DPCs and induce hair follicles in mouse models [13]. The results of experimental studies showed that Wnt1a- and BM-MSC-conditioned media stimulate the conversion of hair follicles from the telogen to anagen phase. This cocktail was also found to significantly increase the number of hair follicles in mice [45]"

Maintaining Hair Inductivity in Human Dermal Papilla Cells: A Review of Effective Methods

[AinmBeo]

@Mauritio
Thanks.
I'm a wee bit confused.
I was partly going on this post, in which you mentioned glucose+niacinamide+ apple polyphenols for hair.
https://bioenergetic.forum/topic/1205/niacinamide-as-effective-as-minoxidil-at-treating-androgenic-hair-loss?_=1743946469122

[Mauritio]

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

Sorry I had missed this post. It's essentially the same study and authors we both dug out there.

No problem .

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

The "spatiotemporal" effects really open up a different angle to determine suitable combinations and timings of different topical substrates. We wouldn't want a limited phase of fully engaged "hair growth bloom" followed up by depletion of the underlying stem cells and exacerbated balding.

Totally agree. Gotta look more into LONG term studies on these substances , although they're probably scarce.

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

We also observed glycolysis-mediated histone acetylation is increased and chemical inhibition of acetyltransferase reduces expression of the signature genes associated with hair induction in active DP. "

Good one! Another reason to use butyrate and other HDACs.
The study even cited 2 other studies showing beneficial effects of HDACs on hair.

https://pubmed.ncbi.nlm.nih.gov/30814580/
https://pubmed.ncbi.nlm.nih.gov/22506014/

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

DP also rely on aerobic glycolysis. So no obvious contradictions between HF and DP so far.

But they study cited another one showing that HFSC rely on ANaerobic glycolisis, generating large amounts of lactate (not pyruvate)
So lactate itself might be wanted and needed for hair (stem cells).. something I never thought I'd say.
Or at least less lactate = less hair stem cells

But is their a cost?
Meaning more stem cells = less actively growing hair.
I think the glutamine study hinted at outer root sheath (ORS) cells reverting back to HFSC, which should reduce the hairs in anagen phases.

Then we're running into the glutamine problem again.
Short term vs. long term effects.

https://pubmed.ncbi.nlm.nih.gov/28812580/#:~:text=Furthermore%2C lactate generation appears to,(Ldha)%20prevented%20their%20activation.

Although this study suggests there is no difference in lactate between people with hair loss vs. No hair loss.
Which at least suggests high lactate there is normal and not necessarily a sign of a broken metabolism .

Squalene, a major component of sebum, was higher in people with hair loss, suggesting higher sebum production.
https://pubmed.ncbi.nlm.nih.gov/37760935/

[Mauritio]

@Mauritio said in Reversal of hair loss in Mice with “sugar gel”:

So lactate itself might be wanted and needed for hair

Here's an interesting review. Under 3.4.3 they talk about lactate and hair loss.

Lactate increases VEGF, leading to new blood vessels and better nutrients supply.
Also, lactate can increase glycogen levels, which increases anagen phase.

https://pmc.ncbi.nlm.nih.gov/articles/PMC11879785/

[Mauritio]

@AinmBeo haidut always says NMN is basically niacinamide, which I don't think is true. Not in hair and not in all of the rest of the body . He's very pro niacinamide so maybe he's a bit biased there.
Not saying Niacinamide doesn't have benefits , hope I'm clear on that.

[AinmBeo]

@Mauritio said in Reversal of hair loss in Mice with “sugar gel”:

Total Liquids:
Water: 70 mL
Ethanol: 20 mL
Propylene Glycol: 10 mL
Total: 100 mL

Does the "Ethanol" listed here imply pure alcohol (no water in the mix)?
I am asking because, if I use 100 proof vodka instead, that is half water, so I would have to adjust the ratio.

[CrumblingCookie]

@Mauritio said:

But they study cited another one showing that HFSC rely on ANaerobic glycolisis, generating large amounts of lactate (not pyruvate)
So lactate itself might be wanted and needed for hair (stem cells).. something I never thought I'd say.
Or at least less lactate = less hair stem cells

But is their a cost?
Meaning more stem cells = less actively growing hair.

• "To assess the role of anaerobic glycolysis and lactate on 3C-HFSC proliferation in our system, we inhibited lactase dehydrogenase using FX11 (Flores et al., 2017) and observed that inhibiting lactate production attenuated proliferation of both HFSCs and ORS progenitors (Figure S4H), but did not affect the proportions of the two populations (Figure 4D). This indicated that, consistent with Flores et al., anaerobic glycolysis is required for HFSC activation, but it does not influence HFSC to ORS lineage progression."

Oh wow. HFSC do require lactate for their own proliferation. But on average it doesn't seem to hinder their absolute differentiation, it seems.

@Mauritio said:

The study even cited 2 other studies showing beneficial effects of HDACs on hair.

https://pubmed.ncbi.nlm.nih.gov/30814580/
https://pubmed.ncbi.nlm.nih.gov/22506014/

Nice follow up on this! I am unsure, however, whether these effects are mediated through HDACi or whether they are separate effects on Akt activation->GSK3beta inhibition->Wnt activation. It may be a distinct mechanism? But indeed why do they list 4-phenyl-butyrate (HDACi) as having the same effects? And trichostatin A (TSA) in the other study (which increased the acetylation level of histone H3K19 and H3K14).
They say VPA inhibits GSK-3beta "indirectly by an as-yet-unknown pathway" but that's info back from 2002. There seem to have been later studies in which HDAC activity was blocked or irrelevant and yet VPA directly inhibited GSK3β enzymatic activity. TSA on the other hand, as in the linked study, doesn't seem to act through Wnt/β-catenin but through increasing bone morphogenetic proteins (BMPs) which sound odd in the context of skin and hair.
(Topical) lithium will also do the Akt activation->GSK3beta inhibition->Wnt activation, but directly.

Even in conditions of sufficient glycolysis the activation of the Wnt/β-catenin pathway apparently lies at the very beginning at DP activation and HF induction because of the presence of overriding other adverse, inhibitory influences.
Which matches my experiences with topical glucose yielding strenghtened hair roots yet no proliferation.

@Mauritio said:

Here's an interesting review. Under 3.4.3 they talk about lactate and hair loss.

Lactate increases VEGF, leading to new blood vessels and better nutrients supply.

... so, topical lactate? If the topical solution remains at pH <5 we may even use benzoic acid or benzoate at 0.5-0.9% as a preservative instead of the high 35% ethanol. Maybe combined with an equimolar amount of glycine to allow for its enzymatic conversion to hippuric acid once applied and absorbed.

@Mauritio said:

haidut always says NMN is basically niacinamide, which I don't think is true. Not in hair and not in all of the rest of the body .

I've ditched all my NAM. I find its effects outright antimetabolic beyond the very short term yet those of nicotinic acid much more agreeable and I've been wanting to source NMN.

[Mauritio]

I've updated tie formula and added sodium butyrate. The amount needed is surprisingly small, and I've already taken a couple times more than needed. This is based on one in vitro study so very speculative, but I've read something about it increasing osmotic stress if you exceed 10mM, not sure if that's true/relevant.

New formula:

D-Glucose: 1%
Apple Peel Polyphenols: 1%
NMN: 1%
Sodium Butyrate: 0.075%
Ethanol: 35%
Water: 65%

[AinmBeo]

@Mauritio
How can one tell if something is D or L glucose?
The makers of this have no clue:
https://www.amazon.com/dp/B097CQ3F71?ref_=ppx_hzsearch_conn_dt_b_fed_asin_title_1

[Gaston]

You guys are cookin with gas now, I can't understand what the hell you're talking about at this point. Keep it up.

[alfredoolivas]

@Gaston I was thinking, thank god I don't have hair loss, so I don't need to dwelve deep into this research. These guys are onto another level lol, much respect.

[Mauritio]

@AinmBeo said in Reversal of hair loss in Mice with “sugar gel”:

Does the "Ethanol" listed here imply pure alcohol (no water in the mix)?

Yes.

[Mauritio]

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

This indicated that, consistent with Flores et al., anaerobic glycolysis is required for HFSC activation, but it does not influence HFSC to ORS lineage progression."

That's good to know.

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

Nice follow up on this! I am unsure, however, whether these effects are mediated through HDACi or whether they are separate effects on Akt activation->GSK3beta inhibition->Wnt activation. It may be a distinct mechanism?

I'm not sure.im not deep into the HDAC literature. I just recall from one of the studies I read today that they said it was am epigenetic mechanism stabilizing the pro-hair genes .

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

so, topical lactate? If the topical solution remains at pH <5 we may even use benzoic acid or benzoate at 0.5-0.9% as a preservative instead of the high 35% ethanol. Maybe combined with an equimolar amount of glycine to allow for its enzymatic conversion to hippuric acid once applied and absorbed.

Lol I've been peating too long to voluntarily apply lactic acid to my scalp, I feel safer with giving my scalp the raw material(s) ,mainly dextrose, and then let it figure it out.

Why do you think it's less than 5 ? I haven't calculated it but I guess it should be higher . Especially once adding potassium bicarbonate. Why benzoic acid?
The ethanol is not just there as a preservative but also as a solvent and absorption enhancer . The apple polyphenols are poorly water soluble IIRC but better soluble in ethanol for example. So benzoic acid would have to be able to do these things as well.

[Mauritio]

@Gaston said in Reversal of hair loss in Mice with “sugar gel”:

You guys are cookin with gas now, I can't understand what the hell you're talking about at this point. Keep it up.

@alfredoolivas said in Reversal of hair loss in Mice with “sugar gel”:

I was thinking, thank god I don't have hair loss, so I don't need to dwelve deep into this research. These guys are onto another level lol, much respect.

Lol thanks guys.
Great teamwork on this thread. This could help many, many people if it works out.

[Mauritio]

Here's a guy on reddit who claims to have cured his balding by using anti fungal spray on his head. Even included some decent pictures.

https://www.reddit.com/r/bald/s/SMe3lriWFf

[CrumblingCookie]

@Mauritio said:

Why do you think it's less than 5 ? I haven't calculated it but I guess it should be higher . Especially once adding potassium bicarbonate. Why benzoic acid?

It's just a thought for avoiding the degree of scalp skin disinfection from the ethanol or its alternative of 20-30% propylene glycol. Of which I cannot say how much significance that bears wrt to the skin microbiome.
What's the minimum ethanol concentration for enhancement of absorption?

• The efficacy of benzoic acid and benzoate as a preservative is dependent on the pH, i.e. a pH <5 would be a requirement for the use of BzOH.
  At low pH BzOH's antibacterial action works through inhibition phosphofructokinase (PFK) and therefore prevention of (an)aerobic fermentation of glucose. PFK being touted the rate limiting enzyme for glycolysis of course urges to keep its topical concentration sensible so that the pH balances out and there'll be no inhibition of glycolysis in the cytoplasm of skin/HF/DP cells.
  Commonly BzOH is <1% (up to 5%) for preservation of cosmetics (limited to 0.5% in the EU and 0.2% in Japan.
• It's easy to confuse with benzyl alcohol, though, which is usually termed BA and used at <1% for preservation of cosmetics because of it being a skin irritant. It needs to be at least 0.9% however as the MIC. BA oxidizes to BzOH.
• I've thought of boric acid as a preservative, too, but I can't get my hands on that one.

@Mauritio said:

I just recall from one of the studies I read today that they said it was am epigenetic mechanism stabilizing the pro-hair genes .

The linked study showing activation of BMP genes by trichostatin A is likely by HDACi. As yet I know nothing about those bone morphogenetic proteins.
The authors wrote:

• BMPs are important in orchestrating tissue architecture by inducing a group of pivotal morphogenetic signals.

• In stem cells, the presence of BMPs in culture has been shown to be crucial for the maintenance of their primitive states

• BMP signaling in DP cells is required for the maintenance of their hair follicle-inductive properties. In the absence of BMP signals, DPs lose their signature characteristics in vitro and fail to generate hair follicles when engrafted with epithelial stem cells in vivo

• In our previous study, we found that BMP2, BMP 4 or BMP6, with BMP4 in particular, could increase the AP [alkaline phosphatase] activity of culture expanded SKPs [multipotent skin-derived precursor cells], and supplementation of BMP4 to the culture increased the hair induction ability of SKPs

Stem cell senescene is being alleviated and their autocrine functions [regulation of a cell's own behaviour by itself] restored by H3K14Ac and H3K19Ac. This definetly targets the HFSC and DP.

In dermatological practice they really should be checking up such markers in hair biopsies instead of the useless "Ah well yes sir/m'lady, the results of laboratory analysis of your painfully plucked hairs are in: They are telogen and falling out and we advice you take this agent of chemical castration, brain neurotransmitter and collagen degeneration" - "Gee, thank you, doc. I better heed this because, boy, I really wouldn't know what to do without your great sophistication!"

@Mauritio said:

Lol I've been peating too long to voluntarily apply lactic acid to my scalp, I feel safer with giving my scalp the raw material(s) ,mainly dextrose, and then let it figure it out.

That guy linked by @Hearthfire who used potassium bicarbonate (antimycotic + for the ion channels instead of minoxidil) also applied diluted acetic acid (white wine vinegar) before and after to enhance scalp perfusion and to (maybe/wishfully) dissolve skin calcium deposits.
Might as well choose lactic acid for that instead? Its use (5%) against acne, rosacea, for stimulating skin collagen and ceramide synthesis and for retaining skin moisture seems very safe and established.
If it were to only benefit the scalp skin even that would be beneficial for the perfusion and nutrient gradient behind the skin barrier. If the skin cells are more stable they wouldn't need to quarrel with HF, DP and stem cells about who get's the glucose and glutamine. I'd consciously deem it wise if skin cells always were to be prioritized over hair.

[Hearthfire]

@Mauritio

That's very interesting. More and more I lean towards hair loss being an occult fungal, parasitic, and possibly bacterial infection in the scalp, Probably enabled by an upstream deficiency. Something happens in the health balance that allows our body to be weakened allowing the infestation. Or perhaps it's just a product of the environment we live in. Terrain theory. When people see raised DHT levels in scalp, they blame baldness on the DHT, but that DHT overload is probably one of your body's measures to repair, trying to regrow the hair that's being destroyed by the fungus/parasites/bacteria. It's like a negative feedback loop that becomes worse and worse over time. DHT grows hair everywhere else on the body. I'm not saying high DHT does not add to the negative effect, but it not being the main factor could be why hair loss is such a mystery to this day, because other more serious underlying factors are doing more damage.

Just as another example of how this could go un-noticed by big medicine; rosacea, Apparently they don't know the cause, but doing a bit of research reading anecdotes online, it seems this might also be caused by demodex mites in most cases. People have cured it by applying ivermectin and tea tree oil. Rosaeca patients tend to have a higher amount of demodex mites, yet mainstream medicine doesn't link the two.

If the cause of a condition as prolific as rosacea is unknown to doctors/skincare professionals, and it turns out to actually be demodex mites, I think the odds of many cases (maybe a majority) of balding in men could be caused by occult fungal/parasitic/bacterial infection. Obviously there's other causes but I'm mainly focused on those caused by (allegedly) DHT.

@CrumblingCookie said in Reversal of hair loss in Mice with “sugar gel”:

That guy linked by @Hearthfire who used potassium bicarbonate (antimycotic + for the ion channels instead of minoxidil) also applied diluted acetic acid (white wine vinegar) before and after to enhance scalp perfusion and to (maybe/wishfully) dissolve skin calcium deposits.
Might as well choose lactic acid for that instead? Its use (5%) against acne, rosacea, for stimulating skin collagen and ceramide synthesis and for retaining skin moisture seems very safe and established.
If it were to only benefit the scalp skin even that would be beneficial for the perfusion and nutrient gradient behind the skin barrier. If the skin cells are more stable they wouldn't need to quarrel with HF, DP and stem cells about who get's the glucose and glutamine. I'd consciously deem it wise if skin cells always were to be prioritized over hair.

Not really wishfully. Totally plausible, given enough applications over time. Vinegar will dissolve an eggshell in 10 hours. It's also anti-fungal. So you're doubling up on the anti-fungal action as well.

[Hearthfire]

@Mauritio

Any reason to not use NAD+ in a formulation like this? Aren't you using NMN because it's a NAD+ precursor? NAD+ is readily available on Amazon for pretty cheap.

[alfredoolivas]

@Hearthfire Not answering for Mauritio, but NAD+ has a molecular mass of 663g/mol, therefore due to the Dalton Rule, it is too big of a molecule too pass through the skin. NMN has a moleculer weight in the 300s, so it can enter and pass the skin.