Effective Methods to Reverse Gray Hair

Dr. Sandeep Bhasin, MBBS, MS

A. Definition and Significance of Graying of Hair

Hair graying, also known as achromotrichia or canities, is a normal aspect of aging. According to reports, 6–23% of people globally will have 50% gray hair by the time they turn 50. Graying typically begins in the mid-30s for Caucasians, the late-30s for Asians, and the mid-40s for Africans.

Role of Melanin in Hair Coloration

Melanin, a pigment produced by specialized cells called melanocytes, is responsible for determining the color of hair. There are two main types of melanin: eumelanin, which produces black or brown hues, and pheomelanin, responsible for red and yellow shades. The relative proportions of these pigments and their distribution within the hair shaft contribute to the wide range of hair colors observed in humans. The cells that make up the hair shaft, known as keratinocytes, are adjacent to the melanocytes in the hair follicles, where melanin is generated. The color of hair is determined by the distribution and quantity of melanin in the hair shaft; darker hair is produced by higher concentrations of melanin.

A hair follicle consists of three layers. The cuticle is the outermost layer; Cortex is the next layer, and Medulla is the innermost layer.

Melanocyte cells are of two distinct type. Melanogenically Active Melanocyte And melanogenically inactive cells.

Inactive melanocytes are present around the arrector pili muscles and peri matrix and do not produce melanin cells. They are considered to be the melanocyte stem cells.

The active melanocytes are present in the infundibulum and hair bulb. It produces and transfers melanocytes to keratinocytes in the cortex and medulla.

The production of melanin depends on the stage of the hair cycle. Human hair has three stages Anagen, Telogen and Catagen. Melanin cells are produced during anagen phase only. Normally, the Anagen phase lasts 3.5 years. The Follicular Melanocytes produce large amount of melanin cells. the follicular melanocytes have larger melanosomes, Golgi Bodies, and Reticulum compared with Epidermal melanosomes.

Melanogenesis is controlled by three main enzymes Tyrosinase, GP75, and Tautomerase.

The important regulators of melanogenesis are MITF, which is Micro Opthalmalmia associated Transcription Factor which controls melanogenesisby acting on Tyrosinase, Tyrosinase Related protein , Dopachrome Tautomerase DCT. The Stem cells are kept in undifferentiated state by PAX3 which suppresses DCT. During anagen Phase the WNT- B Catenin Pathways activate and stimulate differentiation of Stem Cells into melanocytes, which migrate towards the Hair bulb. It is assisted by Alpha MSH, endothelin, KITL in this differentiation. The Melanocyte cell produces Melanin pigment, which migrates into pericortical Keratinocyte.

Mechanisms of Melanogenesis

1.  Activation of Melanocyte Stem Cells:

The process of melanogenesis begins with the activation of melanocyte stem cells located in the bulge region of the hair follicle. Various signaling pathways, including the WNT/β-catenin pathway, regulate the activation and differentiation of these stem cells into mature melanocytes. During the anagen phase of the hair growth cycle, signaling molecules stimulate the proliferation and migration of melanocyte stem cells towards the hair bulb.

2.   Melanin Production and Transfer During the Hair Cycle:

Once activated, melanocytes in the hair bulb begin synthesizing melanin through a series of enzymatic reactions. Enzymes such as tyrosinase, GP75, and dopachrome tautomerase catalyze the conversion of tyrosine into melanin precursors. These precursors are then transferred to neighboring keratinocytes, where they contribute to the pigmentation of the hair shaft. Melanin production is most active during the anagen phase of the hair cycle, when hair growth is at its peak.

In Catagen and telogen phases, melanocyte cells undergo apoptosis and Stem Cell undergo quiescence under the influence of TGF Notch Ligands, DLL1 and WNT inhibitors such as SFRP and DKK3.

Premature graying of the hair is common in present-day society. Its incidence is increasing. Factors associated with it are:

• Family History of Pre Mature Graying of Hair was reported in 65.83% by Sharma et al. and in 75% by Daulatabad et al.

• Increased Oxidative stress as a result of smoking and Pollution

• Increased Mental stress

• Metabolic Disorders Syndrome has a strong association with pre mature graying of the hair.

There are many metabolic disorders linked with Premature Graying of hair. Copenhegan City Heart Study associated Premature Graying Of the hair with myocardial infarction, with a relative risk of 1.9 from secondary data analysis.

Kocaman et al found incidence of hyperlipidemia was higher in patients with Premature Graying of Hair

Shin et al associated increased incidence of Obesity with Premature graying of Hair.

Similarly, Rosen found higher incidence of Osteopenia by 4.4 in Premature Graying of Hair.

Acer et al found 36.5% % patients with premature Graying Of Hair had Atopy. Graying hair with age can be attributed to:

• Decrease in CD +200 Stem cells in Hair follicles

• Increased oxidative stress as in Smoking, UV radiation Exposure and Pollution

• Decreased Catalase enzymes in Hair follicles with age which can protect outer hair shoots against oxidative damage. This increases with Progression of age.

• Reduction in Bcl-2, methionine Sulfoxide reductase and catalase enzyme with age which neutralizes Reactive Oxygen species and Oh group build up in peri follicular area.

• Deficiency of various minerals and vitamins can cause Pre Mature Graying Of Hair

• Cupper needed by Tyrosinase enzyme

• Calcium needed for Phosphorylation and activation

• Ferritin

• Zinc

• Folic Acid

• Vitamin B 12

• • Use of various drugs can cause Graying of Hair as

• Anti epileptics

• Chloroquine

• Hydroxy chloroquine

V. Pharmacological Approaches to Reversing Graying

There have been many reports with various other medications which can cause reversal of graying. These can be categorized into three classes

1. Anti Inflammatory drugs
2. Drugs which increase Melanogenesis
3. Vitamins and Supplements

Anti Inflammatory drugs to cause Regimentation Of Hair

Though hair follicles are immune privileged and are not attacked by immune cells, with time, inflammatory cytokines such as IL6, IL2, TNF -Alpha , etc may build around peri Follicular cells. This may cause Oxidative damage to Melanocytic stem cells and Melanocytes. There is dendritic blunting , defective melanosome transfer, and defective pericortical keratinocytes to receive melanin from melanocytes.  The broken down immune barrier of Hair Follicle allow these inflammatory Cytokines to accumulate in Peri follicular area and inhibit melanocyte functioning.

By using some anti-Inflammatory medicines, there have been case reports of reversal of Graying of hair. The literature shares the following medicines which have reversed graying of hair.

• Tyrosinase Kinase inhibitors such as Imatinib

• Thalidomide, lenalidomide

• Cyclosporin A

• PDL-1 blockers

• Adalimumab

• Acitretin Etretinate

Another group of medicines that can help in the reversal of graying of hair are those drugs that can increase melanogenesis. Along with immunomodulation, it is important to stimulate melanogenesis.

So, these drugs that have been studied so far are:

• Imatinib, erlotinib
• Psoralen
• Thyroxine
• Acitretin
• Tamoxifen
• Leva Dopa
• Latanoprost

Some vitamins, such as Calcium Pantothenate and PABA, have been tried for graying hair.

High-dose p-aminobenzoic acid (PABA) has been tried in canities in various studies. One study mentioned that giving 100 mg PABA 3 times daily produced visible improvement in the form of hair darkening, which relapsed quickly within 2–4 weeks of stopping treatment. In research led by Pasricha Pavithran, calcium pantothenate was also found to be beneficial in preventing premature graying of hair, with full repigmentation occurring in 45% of the instances. Gray hair reversal was noted by Bellandi et al. in individuals receiving latanoprost for an extended period of time. Gray hair repigmentation was documented by Sakhiya et al. after using 5% melitane. Melitane is an alpha-MSH agonist and biomimetic peptide. It functions by binding the melanocortin 1 receptor, which in turn activates the hair’s melanogenesis process. Palmitoyl tetrapeptide-20 is also another alpha-MSH biomimetic peptide. Studies have also shown its beneficial role in promoting hair pigmentation and reversing gray hair.

Role of Cerebrolysin in Hair Repigmentation

Cerebrolysin, a neuropeptide preparation derived from porcine brain, is renowned for its regenerative properties, particularly in neurological applications. Recent studies suggest that Cerebrolysin might also positively affect hair pigmentation by enhancing melanocyte activity and melanin synthesis within hair follicles.

Study Overview

A recent paper published in the European Journal of Medicine in November 2021 highlighted that Cerebrolysin could induce hair repigmentation associated with MART-1/MELAN-A reactivation. The following outlines a case report and a prospective comparative pilot study approved by the ethical committee under protocol number NC-15-001.

Mechanism of Action

Cerebrolysin is a low molecular weight neuropeptide used clinically for various neurological conditions, such as:

• Stroke
• Dementia
• Subarachnoid hemorrhage Alzheimer’s disease Traumatic brain injury
• Arresting neurodevelopmental delay in infants

The drug is believed to enhance neuronal network stability by inhibiting Calpain, increasing neuroplasticity, neurogenesis, neuroprotection, and neurotrophicity. Given that melanin cells derive from neuronal crest cells, cerebrolysin’s activation of neurotrophic factors might influence melanin activation and melanosome biogenesis. The proposed pathways include:

• Activation of RET receptors in stem cells is critical for the stem cell niche microenvironment around the hair follicle
• Activation of P75 neurotrophic factors prevents oxidative damage and apoptosis of the hair follicle’s outer root sheath
• Increased melanosome biogenesis, leading to higher melanin pigment secretion
• Enhanced melanin concentration in hair bulbs by mitigating oxidative damage to stem cells and hair roots

Safety and Side Effects

While cerebrolysin is generally considered safe, some occasional side effects have been reported:

• Vertigo
• Dizziness
• Headache Nausea
• Fever and urinary tract infections
• Hypersensitivity reactions

Case Report and Pilot Study

The pilot study involved five patients already receiving Cerebrolysin for neurological issues (stroke, dementia, or traumatic brain injury). The participants included three males and two females, with a median age of 70 years (ranging from 63 to 77 years).

Hair Whitening Score

The intensity of hair graying was assessed using a Hair Whitening Score:

• All black hair
• More black hair than white hair
• Equal amounts of black and white hair More white hair than black hair
• All white hair

Methodology

To evaluate melanin pigment and melanosome levels in hair follicles, punch biopsies from different areas were studied using:

1. Hematoxylin and eosin staining
2. Fontana-Masson histochemical staining to assess melanin pigment levels
3. Immunohistochemical labeling with mouse monoclonal antibody against Melan-A or MART-1 to detect new melanosome biogenesis

Treatment Protocol

The treatment protocol involved infusing five vials of Cerebrolysin per week:

• Two vials on Monday
• Three vials on Thursday

Each 10 ml vial of Cerebrolysin contained 215.2 mg/ml of the active ingredient, was dissolved in 100 ml of normal saline, and infused over 60 minutes. This cycle was repeated over four weeks, followed by an eight-week break, for a total of three cycles over nine months.

Results

After nine months, reevaluation showed improvements in the morphological appearance of hair, as indicated by the Hair Whitening Score. Additionally, Fontana-Masson staining and immunohistochemical labeling against Melan-A cells demonstrated a four-fold increase in microdensometric measurement of melanin pigment staining. These results suggest that cerebrolysin can enhance melanin synthesis and melanosome biogenesis in hair follicles.

How can we prevent Graying of Hair?

We must understand that the basic problem with graying of hair with age is an overload of oxidative stress. One reason why hair follicles turn gray with age is because there is overload of Hydrogen peroxide free radicles. At younger age, the Hydroxyl ion are neutralized by Catalase enzyme and Methionine Sulfoxide reductase A and B.

So the enzymes that neutralize the Hydroxyl ions are:

• Catalase
• Methionine Sulfoxide Reductase A
• Methionine Sulfoxide Reductase B

With age, Catalase production drops and so does the production of Methionine Sulfoxide enzymes. This causes the buildup of Hydrogen per oxide and Hydroxyl ions. These bleach our melanin pigments and cause Greying of Hair. To overcome this, we can take following supplements:

• Methionine
• N Acetyl Cysteine
• Ubiquinol

If we consume Methionine as a supplement, it protects Catalase from damage but increases our Homocysteine level, so to balance this, whenever we take L Methionine we need to add:

• L- Methyl Folate
• Glycine
• Tri Methyl Glycine

To reduce inflammation, we can add flavonoids such as grape seed Extract and quercetin, which can reduce the damage to Stem cell.

One thing we must understand is that most Permanent Dyes further Damage Hair and increase graying.

One reason to accelerate the graying of hair is the use of permanent dyes. We must understand.

How these permanent Dyes work?

Each Hair Follicle is made up of three parts, cross sectionally:

• Outer most Cuticle
• Middle Cortex
• Inner most medulla

Most permanent dyes work by using Ammonium Hydroxide as a reagent to cause swelling in the cuticles and open up spaces between the cuticle so that the coupler with the dye can enter in the Hair cortex. 

This ammonium hydroxide with hydrogen peroxide used to bleach hair causes damage to the hair and bleaches the hair. Once the cuticles are opened up, we add a Dye in form of paraphenylene Phenylene Diamine PPD, which causes further damage to Hair shaft and Hair roots. PPD can cause delayed allergies and Cancers of various organs, such as Ovary Cancer, Bladder tumors, brain Cancers, and hemopoietic cancers. So permanent dyes are not only harmful to hair but also cause many side effects. Zhang et al reported that permanent dyes are more Hazardous than Semi Permanent Dyes.

• Damage to Hair Root
• Bleaching and Damage to Hair Melanin Pigments
• Allergic Reaction
• Irritant dermatitis
• Contact Dermatitis
• Cancers in Long Run
• Further cause Graying of Color

So permanent dyes perpetuate the Graying of Hair. We can substitute these with Semi Permanent Dyes which don’t damage hair cuticles using Ammonia and PDP free dyes, shifting to Herbal Colors as Henna and Indigo.

Stress and Graying of Hair

Hair follicles are surrounded by Sensory and Sympathetic nerve fibers. These are present around erector Pilli Muscle and Hair Bulb. Under stress and Sympathetic overactivity, the excess secretion of Nor Adrenalin can cause excess proliferation of Melanocytic Stem Cells which proliferate rapidly and undergo exhaustion. So stress causes depletion of stem cells responsible for our Hair pigmentation There are studies going on to use P27, which is inhibitor of Cyclin dependent Kinase, which can prevent stress induced Graying Of Hair.

How can we Bio Hack Graying of Hair?

• Avoid use of Permanent Dyes. Use Herbal dyes such as Henna and Indigo
• Take nutraceuticals such as Ubiquinol, Vitamin B12, Zinc, Methionine, N – Acetyl Cysteine , Glycine etc.
• Add some anti-inflammatory supplements, such as Grape Seed Extracts and Quercetin
• Avoid smoking, pollution and UV radiation exposure
• Reduce stress by meditation, yoga Mindfulness
• Cerebrolysin Injections can be quite useful, along with above measures
• Topical use of Minoxidil, cyclosporine and Tacrolimus
• Topical use of Alpha MSH agonists as Melitane and Palmityl Tetrapeptide 20

Combination therapies that target multiple pathways involved in melanin production and transfer may offer synergistic effects and improve treatment outcomes.

Conclusion

The pilot study shows cerebrolysin may reverse hair graying by boosting melanocyte activity and melanin production. While this is promising for hair repigmentation treatments, more research is needed to confirm these results and assess their long-term safety. Future studies, like those by Rivertown Therapeutics and the University of Alabama at Birmingham, are investigating combination treatments to preserve melanocyte stem cells and reduce inflammation, which may offer new insights into preventing and reversing hair graying.

In summary, cerebrolysin and similar treatments hold potential for addressing hair graying by activating neurotrophic factors and enhancing melanin synthesis in hair follicles.

References:

1. Paus R, Müller-Röver S, Van Der Veen C, et al. A comprehensive guide for the recognition and classification of distinct stages of hair follicle morphogenesis. J Invest Dermatol. 1999 Dec;113(6):523-32. doi: 10.1046/j.1523-1747.1999.00737.x. PMID: 10594741.
2. Tobin DJ. The cell biology of human hair follicle pigmentation. Pigment Cell Melanoma Res. 2011 Feb;24(1):75-88. doi: 10.1111/j.1755-148X.2010.00745.x. Epub 2010 Nov 9. PMID: 21062352.
3. Picardo M, Dell’Anna ML, Ezzedine K, et al. Vitiligo. Nat Rev Dis Primers. 2015 Oct 22;1:15011. doi: 10.1038/nrdp.2015.11. PMID: 27189774.
4. Hillebrand GG, Levine MJ, Miyamoto K, et al. Bioengineering of human skin: water and the stratum corneum. Crit Rev Ther Drug Carrier Syst. 1991;7(2):135-56. PMID: 1812986.
5. Inoue-Narita T, Hamada K, Sasaki T, et al. Pten deficiency in melanocytes results in resistance to hair graying and susceptibility to carcinogen-induced melanomagenesis. Cancer Res. 2008 May 15;68(10):5760-8. doi: 10.1158/0008-5472.CAN-07-6506. PMID: 18483266.
6. Sasaki M, Horikoshi T, Uchiwa H, et al. Up-regulation of tyrosinase gene by nitric oxide in human melanocytes. Biochem Biophys Res Commun. 2000 Jul 5;273(2):481-4. doi: 10.1006/bbrc.2000.2985. PMID: 10873693.
7. Cerebrolysin induces hair repigmentation associated to MART-1/Melan-A reactivation Gustavo Villarreal-Reyna,#1 Rodolfo Garza-Morales,#2 Adolfo Soto-Domínguez,2 Lorena Montañez- Guerrero,1 Odila Saucedo-Cárdenas,2 Minerva Gómez-Flores,3 Jorge Alejandro Ocampo-Garza,3 José Juan Pérez-Trujillo,2 and Roberto Montes-de-Oca-Lunacorresponding author2