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Effects of tocotrienol supplementation on hair growth in human volunteers

Tropical Life Sciences Research, 21(2), 95–103, 2010
Effects of Tocotrienol Supplementation on Hair Growth in Human
1Lim Ai Beoy, 2Wong Jia Woei, 1Yuen Kah Hay
1School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800, Pulau Pinang,
2Hovid Bhd, 121, Jalan Tunku Abdul Rahman, 30010 Ipoh, Perak.

Studies have shown an association between oxidative stress and alopecia.
Patients with alopecia general y exhibit lower levels of antioxidants in their scalp area as
well as a higher lipid peroxidation index. Tocotrienols belong to the vitamin E family and
are known to be potent antioxidants. Hence, a study was conducted to investigate the
effect of tocotrienol supplementation on hair growth in volunteers suffering from hair loss.
Twenty-one volunteers were randomly assigned to orally receive 100 mg of mixed
tocotrienols daily while seventeen volunteers were assigned to receive placebo capsule
orally. The volunteers were monitored for the number of hairs in a pre-determined scalp
area as well as the weight of 20 strands of 1-cm length hair clippings at 0 (before
supplementation), 4 and 8 months. The number of hairs of the volunteers in the tocotrienol
supplementation group increased significantly as compared to the placebo group, with the
former recording a 34.5% increase at the end of the 8-month supplementation as
compared to a 0.1% decrease for the latter. Nevertheless, the cumulative weight of 20
strands of hair clippings did not differ much from the baseline for both supplementation
groups at the end of the study period. In conclusion, this trial demonstrated that
supplementation with tocotrienol capsules increases hair number in volunteers suffering
from hair loss as compared to the placebo group. This observed effect was most likely to
be due to the antioxidant activity of tocotrienols that helped to reduce lipid peroxidation
and oxidative stress in the scalp, which are reported to be associated with alopecia.

Keywords: Hair Growth, Tocotrienols, Antioxidants
Scalp hair plays an important function in humans. In addition to providing cranial
cushioning and shielding the scalp from direct sun rays, hair has sociological
meanings in terms of gender, age, values and status (Cash, 2001). Scalp hair is
also a vital aspect of an individual physical appearance and helps to project a
positive image (Cash, 2001). Hair loss and balding, on the other hand, are
associated with negative attributes (Rushton et al, 2002). Hair loss or alopecia is
a common problem suffered by both sexes and has been reported to cause
significant psychological effects, such as diminished self-esteem, emotional
distress, embarrassment and social inadequacy, with these detrimental effects
being more significant in women (Cash et al, 1993). Donk et al (1994) observed
that alopecia could severely affect the quality of life of the majority of the
sufferers, whereby 88% of sufferers experienced negative effects in their daily life
and 75% and 50% encountered negative self-esteem and social problems, respectively. There are many types of alopecia, such as androgenetic alopecia (AGA), alopecia areata, telogen effluvium, hair loss due to systemic medical problems such as thyroid disease and adverse drug effects, as well as hair loss due to scalp or hair trauma, discoid lupus erythematosus, lichen planus and structural shaft abnormalities, to name a few (Hogan and Chamberlain, 2000). The causes for the above are numerous and vary between the different types of alopecia. In most instances, the aetiology is still unknown. However, one of the universal causes of increased hair thinning or shedding is nutritional deficiency. It was previously reported that children with protein malnutrition would exhibit fragile and finer hairs that easily fall or break off and display a lower daily rate of growth (Bradfield & Bailey, 1969; Sims, 1968). Nutritional factors that have been identified to be essential in preventing hair loss include iron and an essential amino acid, L-lysine (Rushton et al, 1990; Rushton, 2002). Rushton et al (2002) reported that women with chronic telogen effluvium have low serum ferritin levels and proceeded to demonstrate that daily supplementation of iron and L-lysine for 6 months could cause significant increases in hair numbers. Recently, Naziroglu et al (2000) showed that there was an association between oxidative stress and alopecia. They reported that the levels of reduced glutathione (GSH) and activities of glutathione peroxidase (GSH-Px), which are present to protect against damages caused by reactive oxygen species such as free radicals and peroxides, were significantly lower in patients with alopecia than in controls, whereas the levels of thiobarbituric acid reactive substances (TBARS), which indicate lipid peroxidation and oxidative stress, were significantly higher. Tocotrienols together with tocopherols are members of the Vitamin E family. They share similar structural features of a chroman head and a 16-carbon phytyl chain. The structural difference between them lies mainly in the latter possessing a saturated phytyl chain whereas that of the former possesses three unsaturated double bonds (Papas, 1999; Theriault et al, 1999). Tocotrienols also possess more potent antioxidant property, and Serbina et al (1991) showed that α-tocotrienol was 40- to 60-fold more potent than α-tocopherol against lipid peroxidation in rat liver microsomal membranes. This superiority has been ascribed to the ability of tocotrienols to better distribute within the fatty layers of the cell membranes and hence permit better interaction with lipid radicals. Moreover, tocotrienols have been shown to afford protection to the skin against UV light- and ozone-induced oxidative stress (Traber et al, 1997). In view of the association between alopecia and oxidative stress and the high antioxidant potency of the tocotrienols, a study was thus performed in volunteers with alopecia to evaluate the efficacy of tocotrienol supplementation in improving hair coverage of the scalp and preventing hair thinning in patients suffering from alopecia. Effects of Tocotrienol Supplementation on Hair Growth METHODOLOGY

Patient Population

Thirty-eight male and female volunteers ranging from 18 to 60 years old who met
the inclusion criteria were recruited into the trial. Volunteers had varying levels of
hair loss, ranging from patchy loss of scalp hair to more severe loss of scalp hair.
Hair loss must have been present for at least 2 months and the alopecia area
could not have any visual evidence of new hair growth. All volunteers were in
good general health with no evidence of systemic illness including cardiac,
psychiatric, thyroid and scalp diseases.
Volunteers were excluded from the study if they had known hypersensitivity to tocotrienols or if they had undergone hair restoration procedures, consumed hair growth medications or applied hair enhancement products in past six months. Patients who had undergone chemotherapy and experienced scalp trauma were also excluded. Concomitant use of medications, such as steroids, oral contraceptives, cytotoxic agents, vasodilators, antihypertensive agents, anti-convulsant drugs, ß-adrenergic receptor blockers, diuretics, spironolactone, cimetidine, cyclosporine or ketoconazole, during the six months prior to the study was also not allowed. Patients on iron and vitamin B12 supplementation were also excluded. Volunteers were also instructed not to alter their hairstyle, the hair care products (such as shampoo or conditioner) currently in use or dye their hair
during the study period.
Study Design
This study was an 8-month (32-week), randomised, double-blind, placebo-
controlled trial conducted at the School of Pharmaceutical Sciences, Universiti
Sains Malaysia. The protocol and informed consent form were approved by an
institutional review board and written informed consent was obtained from each
volunteer prior to enrolment into the study. Thirty-eight volunteers participated in
the trial, and twenty-one volunteers were randomly selected to receive tocotrienol
supplementation (tocotrienol supplementation group) while seventeen were
randomly selected to receive the placebo (placebo group). The placebo is a soft
gelatine capsule containing 600 mg of soya bean oil (Hovid Sdn Bhd, Ipoh,
Malaysia), while tocotrienol supplementation consisted of mixed tocotrienols
formulated in a soft gelatine capsule (Hovid Sdn Bhd). Each tocotrienol capsule
contained 50 mg of mixed tocotrienols (30.8% α-tocotrienol, 56.4% γ-tocotrienol
and 12.8% δ-tocotrienol) as well as 23 IU of α-tocopherol. The volunteers were
required to take one capsule of either the placebo or tocotrienol twice daily after
meal over a period of eight months. Hence, the total daily intake of tocotrienols
for each volunteer in the treatment group was 100 mg. The capsules were
provided on a monthly basis and compliance with the supplementation regimen
was monitored by counting the capsules that were left over during the monthly
visit. The volunteers were also counselled and advised to take the capsules
regularly as per the instruction given to ensure compliance. The volunteers were
required to report any side/adverse reactions, additional medication taken and
any new diagnosis during the trial period.
Efficacy Evaluation

Two parameters were chosen to evaluate the efficacy of tocotrienol
supplementation (Price et al, 2002):
1. Hair
Hair counts served as the primary efficacy measure. An area of 2 × 2 cm was selected in the area of hair thinning for each patient in the balding vertex scalp, and the two opposing corners of the square were permanently marked (using a 4-cm2 wire template) to ensure that the hair in same area was counted at each visit. The count was done by an investigator who was blinded to the treatment the volunteers were receiving. The resulting hair counts were used to calculate the percentage change from baseline. A small tuft of hair (at least 20 strands) within the demarcated area was clipped horizontally. Twenty strands were randomly chosen and cut to 1 cm in length. The total weight was obtained using a microbalance and the resulting mean weight was used to calculate the percentage change from baseline. Both parameters were obtained at baseline, 4 months (16th week) after the start of the study and at the end of 8 months (32nd week) for efficacy evaluation.
Altogether, there were three intervals.
Statistical Calculation

The data collected at baseline and during the supplementation period for both
groups were compared using an analysis of variance procedure (ANOVA) for a
two-factor repeated measures split-plot experimental design (Kirk, 1968). If a
statistically significant difference for the main effect of interval was observed,
Tukey’s post-hoc test was employed. When significant interaction between
supplementation and an interval was observed, simple main effect analysis
(pairwise comparison) was employed to locate the pair that gave rise to the
observed difference. The homogeneity of the baseline characteristics between
the two groups, namely the number and weight of hairs, was also assessed. A
statistically significant difference was considered at p < 0.05.

Thirty-eight volunteers with signs of hair loss or balding were enrolled in the
study, with the supplementation group having twenty-one volunteers (19 male
Effects of Tocotrienol Supplementation on Hair Growth and 2 female) and the placebo group (controlled group) having seventeen volunteers (all male). The age of the study population ranged from 18 to 59 years. A wide range in the age of volunteers was observed because hair loss is known to affect a wide range of the population irrespective of age. Although it is more commonly seen with increasing age, young adults are also affected. Moreover, hair loss may be attributed to universal causes such as nutritional deficiency and oxidative stress, which can affect a wide age group. The volunteers had hair loss problem for approximately 2 to 5 years. Comparability of the treatment groups with respect to initial hair counts as well as the weight of hair prior to study commencement was also assessed. The mean number of hairs of volunteers in the placebo group at baseline was 289.0 ± 98.3, while that of the tocotrienol supplementation group was 284.8 ± 111.3. As for the mean weight of 20 strands of hair at baseline for the placebo and tocotrienol supplementation groups, the values were 0.1002 ± 0.0639 g and 0.0920 ± 0.0565 g, respectively. The values between the placebo and supplementation groups were comparable for both parameters and no statistically significant difference (p > 0.05) was detected. Thirty-five volunteers completed the entire study and were included in the evaluation of the efficacy of tocotrienol supplementation. Three volunteers were lost to follow-up, one (female) from the tocotrienol supplementation group and two from the placebo group. Compliance with the supplementation regimen was satisfactory, as evidenced by the capsule counts during their monthly visits. No severe drug reaction related to the administration of the drug product and placebo was recorded, thus indicating that long-term administration of 100 mg of tocotrienols for up to 8 months was tolerable. The compliance with the study protocol was also deemed satisfactory and no protocol deviation was recorded. Table 1 shows the hair numbers at baseline and 4 and 8 months from baseline for volunteers receiving placebo and tocotrienol supplementation together with the percentage change from baseline. It is evident that volunteers in the tocotrienol supplementation group showed a gradual mean increase in hair numbers from baseline up to the eighth-month interval with a mean percentage increase of more than 34% at the end of the study. In comparison, the number of hairs for volunteers in the placebo group did not show any appreciable increase at the end of the 8-month study. A negligible percentage change of 0.1% was recorded by this group. There was no statistically significant difference in the main effect of supplementation but a significant main effect of interval was evident. Tukey’s post-hoc test showed that the hair numbers at the 8-month interval were significantly greater than those at baseline and at the 4-month interval. Subsequent simple main effect analysis indicated that a statistically significant difference occurred between baseline and post-supplementation at the 8-month interval for volunteers supplemented with tocotrienols only. All other pairwise comparisons (baseline vs. 4-month interval for placebo and supplementation group and baseline vs. 8-month interval for placebo group) did not show statistically significant differences. This indicates that there was no placebo effect and the increase in the number of hairs observed in the volunteers could be ascribed to tocotrienol supplementation. Pairwise comparison also demonstrated that the hair numbers of volunteers of the tocotrienol supplementation and placebo groups were significantly different at the 8-month interval. At this interval, the number of hairs was much higher for the tocotrienol supplementation group volunteers than those in the placebo group. Nevertheless, the same could not be ascribed for observations obtained at the 4-month interval, whereby no statistically significant difference was observed between the two groups at this interval. In this regard, it can be summarised that tocotrienol supplementation for a period of 8 months resulted in significant increases in the numbers of hairs as compared to that in the placebo group. All volunteers with the exception of one in the tocotrienol supplementation group showed a positive response at the end of the 8-month study, recording an increase in the number of hairs in the area of scalp evaluated. Eight volunteers (40.0%) showed hair increases of more than 50%, one volunteer (5.0%) had a 25% to 50% increase, nine volunteers (45.0%) had increases between 10% to 25% while one volunteer (5.0%) showed a hair increase of less than 10%. Only one volunteer (5.0%) in the tocotrienol supplementation group had a slight decrease in the number of hairs. On the other hand, only eight volunteers in the placebo group showed an increase in the number of hairs after 8 months, with one (6.7%) showing more than 20% increase and the remaining seven (46.7%) showing negligible increases. Seven volunteers (46.7%) had a decrease in the number of hairs. However, there was no statistically significant increase in the weight of hair (p > 0.05) between pre- and post-supplementation for both groups of volunteers (tocotrienol supplementation and placebo group). The mean percentage of weight increment after eight months of supplementation was 5.9% in the tocotrienol supplementation group, while that of the placebo group had a slight decrease of 2.0% (Table 2). Recently, Naziroglu et al (2000) showed that an association existed between oxidative stress and alopecia. Specifically, the levels of GSH and activities of GSH-Px were significantly lower in patients with alopecia than in controls, whereas TBARS levels, which indicate lipid peroxidation and oxidative stress, were significantly higher. Similar observations were also reported by Koca et al (2005) and Akar et al (2002), whereby the authors in the former observed an increased lipid peroxidation and a decrease in superoxide dismutase (SOD) levels in patients with alopecia areata compared to controls. In the latter, TBARS levels in the scalp of patients with alopecia areata were higher than those of controls, although the authors reported a higher levels of SOD and GSH-PX in alopecia patients. Girat et al (1996) similarly reported that TBARS levels increased 2-fold and glutathione content was reduced 2.5-fold in sebaceous gland-enriched scalp skin of men affected by male pattern baldness as compared to the controls. Moreover, Naziroglu et al (2000) demonstrated that both the ß-carotene and α-tocopherol levels in plasma were lower in patients with alopecia, albeit the latter was not statistically significant. This led to the authors’ postulation that antioxidant treatment such as using ß-carotene, vitamin E and selenium may benefit patients with alopecia. Hence, the positive effect of tocotrienol Effects of Tocotrienol Supplementation on Hair Growth supplementation in the current study could most likely be ascribed to its potent anti-oxidant activities. Nevertheless, other mechanisms merit further investigations such as those that play a role in AGA. It has been shown by Hee et al (2008) that
incubation of a synthetic androgen with rat vibrissae dermal papilla cells (DP6)
led to increased production of reactive oxygen species (ROS) intracellularly. In
addition, the authors also reported that androgen-inducible transforming growth
factor beta 1 (TGF-ß1), a key mediator in the formation of AGA, was mediated by
ROS and could be prevented by antioxidants or ROS inhibitors in the hair follicle
dermal papilla cells (Hee et al, 2008, Inui et al, 2003). The above observations
led the authors to suggest that antioxidants could potentially be used to control
androgen-mediated pattern hair loss.
In conclusion, this trial demonstrated that supplementation with tocotrienol
capsules increases hair numbers in volunteers suffering from hair loss as
compared to the placebo group. A possible explanation for the effects could be
due to the potent antioxidant activity of tocotrienols that help to reduce lipid
peroxidation and oxidative stress in the scalp, which are known to be associated
with alopecia.

Akar A, Arca E, Erbil H, Akay C, Sayal A and Gur A R. (2002). Antioxidant enzymes and
lipid peroxidation in the scalp of patients with alopecia areata. J Dermatological Sci 29: 85-90. Bradfield R B and Bailey M A. (1969). Hair root response to protein malnutrition. In: Montagna W and Dobson R L (eds). Biology of skin, Hair growth, Vol IX; VII. Oxford: Pergamon Press, 109-119. Cash T F, Price V H and Savin R C. (1993). Psychological effects of androgenetic alopecia on women: comparisons with balding men and with female control subjects. J Am Acad Dermatol 29: 568-575. Cash T F. (2001). The psychology of hair loss and its implications for patient care. Clinics Donk J van der, Hunfeld J A M, Passchier J, Knegt-Junk K J and Nieboer C. (1994). Quality of life and maladjustment associated with hair loss in women with alopecia androgenetica. Social Science & Medicine 38(1): 159-163. Girat M, Cervello I, Nogues M R, Puerto A M, Ortin F, Argany N and Mallol J. (1996). Glutathione, glutathione S-transferase and reactive oxygen species of human scalp sebaceous glands in male pattern baldness. J Invest Dermatol 107: 154-158. Hee C E. (2008). The involvement of ROS on androgen inducible TGF beta 1 regulation derived from dermal papilla cells; a suggestive implication of ROS on androgenetic alopecia. J Am Acad Dermatol 58(2) Suppl 2: AB84. Hogan D J and Chamberlain M. (2000). Male pattern baldness. South Med J 93(7): 657- Koca R, Armutcu F, Altinyazar H and Gurel A. (2005). Evaluation of lipid peroxidation, oxidant/antioxidant status and serum nitric oxide levels in alopecia areata. Med Sci Monit 11(6): CR296-299. Papas AM (1999). Vitamin E: Tocopherols and tocotrienols. In: Papas A M (ed.). Antioxidant status, diet, nutrition and health. Boca Raton, Fl: CRC Press, 189-210. Price V H, Menefee E, Sanchez M, Ruane P and Kaufman K D (2002). Changes in hair weight and hair count in men with androgenetic alopecia after treatment with finasteride, 1 mg daily. J Am Acad Dermatol 46: 517-523. Rushton D H, Norris M J, Dover R and Busuttil N. (2002). Causes of hair loss and the developments in hair rejuvenation. Int J Cosmet Sci 24: 17-23. Rushton D H, Ramsay I D, James K C, Norris M J and Gilkes J J H. (1990). Biochemical and trichological characterization of diffuse alopecia in women. Br J Dermatol 123: 187-197. Rushton D H. (2002). Nutritional factors and hair loss. Clin & Exp Dermatol 27: 400-408. Serbina E, Kagan V, Han D and Packer L. (1991). Free radical recycling and intramembrane mobility in the antioxidant properties of alpha tocopherol and alpha tocotrienol. Free Radic Biol Med 10(5): 263-275. Sims R T. (1968). The measurement of hair growth as an index of protein synthesis in malnutrition. Br J Nutr 22: 229-236. Theriault A, Chao J T, Wang Q, Gapor A and Adeli K. (1999). Tocotrienol: A review of its therapeutic potential. Clin Biochem 32(5): 309-19. Traber M G, Podda M, Weber C, Thiele J, Rallis M and Packer L. (1997). Diet-derived and topically applied tocotrienols accumulative in skin and protect the tissue against ultraviolet light-induced oxidative stress. Asia Pacific J Clin Nutr 6(1): 63-67. Effects of Tocotrienol Supplementation on Hair Growth Table 1. Mean numbers of hairs at baseline and after 4 months and 8 months of
tocotrienol and placebo supplementation (Mean ± SD, percentage change from baseline).

Table 2. Mean weight (gram) of 20 strands of hair at baseline and after 4 months and 8
months of tocotrienol and placebo supplementation (Mean ± SD, percentage change from


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