Anagen Phase Research Articles

Ectodysplasin-A2 induces apoptosis in cultured human hair follicle cells and promotes regression of hair follicles in mice

Ectodysplasin is a ligand of the TNF family that plays a key role in ectodermal differentiation. EDA-A1 and EDA-A2 are two isoforms of ectodysplasin that differ only by the insertion of two amino acids and bind to two different receptors, ectodysplasin A receptor (EDAR) and ectodysplasin A2 receptor (EDA2R), respectively. Mutations of EDA-A1 and its receptor EDAR have been associated with hypohidrotic ecodermal dysplasia (HED). However, the role of EDA-A2 and the expression pattern of EDA2R in human hair follicles and in the mouse hair growth cycle have not been reported. In this study, we first investigated the expression of EDA2R in human hair follicles and in cultured follicular cells. EDA2R was strongly expressed in outer root sheath (ORS) cells and weakly expressed in dermal papilla (DP) cells. EDA-A2 induced the apoptosis of both ORS cells and DP cells via the activation of cleaved caspase-3. In addition, EDA2R was highly expressed in the late anagen phase compared with other phases in the hair growth cycle. Moreover, EDA-A2 induced apoptosis in cultured human hair follicle cells and in the mouse hair growth cycle, causing the premature onset of the catagen phase. Collectively, our results suggest that EDA-A2/EDA2R signaling could inhibit hair growth, and an inhibitor of EDA-A2/EDA2R signaling may be a promising agent for the treatment and prevention of hair loss.

Italian Guidelines in diagnosis and treatment of alopecia areata.

Alopecia areata (AA) is an organ-specific autoimmune disorder that targets anagen phase hair follicles. The course is unpredictable and current available treatments have variable efficacy. Nowadays, there is relatively little evidence on treatment of AA from well-designed clinical trials. Moreover, none of the treatments or devices commonly used to treat AA are specifically approved by the Food and Drug Administration. The Italian Study Group for Cutaneous Annexial Disease of the Italian Society of dermatology proposes these Italian guidelines for diagnosis and treatment of Alopecia Areata deeming useful for the daily management of the disease. This article summarizes evidence-based treatment associated with expert-based recommendations.

Nuclear localization of Meis1 in dermal papilla promotes hair matrix cell proliferation in the anagen phase of hair cycle

The dermal papilla (DP) is a key mesenchymal compartment of hair follicles that orchestrates mesenchymal-epithelial interaction regulating hair growth cycles. In the present study, we demonstrate that a TALE-family transcription factor, Meis1, is selectively localized in the nucleus of the DP in the anagen phase of the hair cycle. By using an ex vivo organ culture of vibrissae follicles, conditional Meis1 loss causes retardation in hair growth, accompanied by defects in cell proliferation of hair matrix cells. This cell proliferation defect is partly rescued by the addition of culture supernatants derived from Meis1-sufficient but not -deficient DP cells. These findings indicate that nuclear Meis1 in DP activate genes involved in secretion of some unknown factors, which promote proliferation of hair matrix cells in the anagen phase of the hair cycle.

Primeira geração de fito peptídeos de Cúrcuma longa mais blend de nutrientes associados a carboxiterapia para o tratamento de Alopecia androgenética masculina

Nos dias atuais, o mercado de estética vem expandindo suas áreas e aprimorando suas técnicas. Uma dessas áreas que vem ganhando força nesse ramo é a terapia capilar que previne e trata as disfunções do couro cabeludo, na busca de minimizar agressões externas e reparar eventuais desequilíbrios fisiológicos. Dentre as disfunções do couro cabeludo podemos citar a Alopecia Androgenética que segundo Banka N, et al (2013) e Olsen EA et al (2003) é quando ocorre miniaturização folicular e encurtamento da duração da fase anágena dos fios acometidos, obtendo como consequência o aumento do número de fios telógenos, resultando em fios mais finos. Trata-se de uma condição hereditária e andrógeno-dependente. O presente estudo de caso associou o uso do ativo inovador Capilia Longa® cuja a ação é reativar o crescimento e reduzir a queda reiniciando e nutrindo o folículo capilar, integrado ao uso da carboxiterapia que é uma técnica, onde se utiliza o gás carbônico medicinal (dióxido de carbono ou CO2) injetado no tecido intradérmico, estimulando os efeitos fisiológicos, como a melhora da circulação e oxigenação tecidual. O presente estudo de caso tem por objetivo avaliar os efeitos da carboxiterapia associado ao ativo capilia longa como resposta para o crescimento e fortalecimento dos fios da fase anágena na alopecia androgenética masculina.

EFFICACY AND SAFETY OF STANDARDIZED CINNAMON BARK EXTRACT FOR THE PREVENTION OF CHEMOTHERAPY-INDUCED WEIGHT LOSS AND ALOPECIA IN PATIENTS WITH BREAST CANCER: A RANDOMIZED, DOUBLE-BLIND, AND PLACEBO-CONTROLLED STUDY

Objective: The objective of the study was to evaluate the effects of IND02 (standardized Cinnamon bark extract) supplementation for the prevention of side effects of cancer chemotherapy in female patients with breast cancer.
 Methods: The study was conducted using double-blind, placebo-controlled design in 34 female breast cancer patients during the first 4 consecutive 21-day cycles of the standard chemotherapy regimen. The active treatment (IND02 capsules, 400 mg, one capsule, and thrice a day) or matching placebo was orally administrated in randomized (1:1 ratio) patients. The efficacy outcome measures were reduction in chemotherapy-induced weight loss, alopecia (hair fall), and other side effects. The safety outcome measures were hematology, ECG, vital signs, adverse event monitoring, and laboratory safety measurements.
 Results: The patients on the treatment with IND02 had shown significant protection from chemotherapy-induced severe weight loss (cachexia) and alopecia (reduced hair density and % hairs in the anagen phase, and increased % hairs in telogen phase) which was seen in the placebo group. IND02 treatment was found safe and well-tolerated during the study.
 Conclusion: Concomitant use of IND02 in breast cancer patients during breast cancer chemotherapy showed a clinical promise regarding efficacy and safety in preventing chemotherapy-induced weight loss and alopecia.

DHT and E2 synthesis-related proteins and receptors expression in male yak skin during different hair follicle stages

Dihydrotestosterone (DHT) and 17β-estradiol (E2) are sex hormones that regulate human hair follicle (HF) growth and are produced by peripheral reduction and aromatization of testosterone. However, the expression patterns of DHT and E2 synthesis-related proteins and their receptors in male yak skin during different HF stages (telogen, anagen, and catagen) are unknown. In this study, we found that both 5α-red and androgen receptor (AR) were expressed in epithelial cells and AR was expressed in the dermal papilla. Additionally, the transcription level of 5α-red1 at different HF stages was significantly higher than that of 5α-red2 mRNA at the same stage; 5α-red1 and 5α-red2 proteins peaked during the anagen and telogen periods of HF, respectively. However, AR protein was only expressed in the skin during the anagen phase of HF. Aromatase and estrogen receptors (ERα and ERβ) were expressed in cutaneous epithelial cells, whereas ERα and ERβ were expressed in the dermal papilla; the transcription level of ERα in HFs at each stage was much higher than that of ERβ. From the catagen to telogen phase, aromatase protein expression was down-regulated, while ERα protein expression was up-regulated. Based on our results, we speculate that 5α-red1 is essential for the synthesis of DHT in male yak skin epithelial cells and promotes the growth of HFs through AR. E2 synthesized by male yak skin epithelial cells may inhibit the growth of male yak skin HFs by ERα. These results provide a foundation for further study on the mechanism of hormone-regulated male yak skin HFs.

Do Kimchi and Cheonggukjang Probiotics as a Functional Food Improve Androgenetic Alopecia? A Clinical Pilot Study.

PurposeProbiotic supplementation demonstrates beneficial effects on serum lipid profiles. We hypothesized that probiotics could benefit patients presenting with alopecia, secondary to improved blood flow to the scalp.Materials and MethodsOur study included men with stage II to V patterns of hair loss based on the Hamilton–Norwood classification and women with stage I to III patterns of hair loss based on the Ludwig classification. All patients were administered 80 mL of Mogut® (a kimchi and cheonggukjang probiotic product) twice a day. Hair growth and numbers were measured using the Triple Scope System® (KC Technology, Korea) at baseline and after 1 and 4 months of administration of a kimchi and cheonggukjang probiotic product.ResultsAt baseline, the mean hair count was 85.98±20.54 hairs/cm2 and the mean thickness was 0.062±0.011 mm in all patients (n=46). Hair count and thickness had significantly increased at 1 month (90.28±16.13 hairs/cm2 and 0.068±0.008 mm, respectively) and at 4 months (91.54±16.29 hairs/cm2 and 0.066±0.009 mm, respectively). In this study, we found that a kimchi and cheonggukjang probiotic product could promote hair growth and reverse hair loss without associated adverse effects such as diarrhea.ConclusionsWe suggest that the observed improvements in hair count and thickness resulted from initiation of the anagen phase in hair follicles in response to probiotics.

Nerve growth factor regulates the proliferation of cashmere goat outer root sheath cells through the activation of cAMP-binding protein

The quality and yield of cashmere are closely related to the growth and development of the secondary hair follicles of cashmere goats. Outer root sheath cell (ORSC) proliferation is considered particularly important for the growth and development of hair follicles; however, the factors affecting this are unclear. We previously showed that the expression of nerve growth factor (NGF) in secondary follicle ORSCs was significantly higher in the anagen phase than in the catagen phase, suggesting that NGF promotes ORSC proliferation. To investigate whether ORSCs secrete NGF, and to investigate its role in ORSCs from goat secondary hair follicles, we used enzyme linked immunosorbent assay (ELISA) to show that NGF levels in cultured ORSC medium decreased significantly with the passage of time (P < 0.05). Immunofluorescence analysis detected the transcription factor cAMP-binding protein (CREB) in ORSCs, with a stronger signal intensity observed in the nucleus than in the cytoplasm. An alkaline phosphatase reporter system assay showed that the exogenous addition of 20 ng/mL or 100 ng/mL recombinant NGF protein significantly enhanced the activity of CREB in ORSCs, while treatment with K252α, which inhibits tropomyosin receptor kinase A (TrkA), revealed that NGF enhanced intracellular CREB activity in an NGF/TrkA-dependent manner. Real-time quantitative PCR investigated the expression of proliferating cell nuclear antigen (PCNA) mRNA after treatment with NGF or the CREB inhibitor ICG-001. The NGF/TrkA/CREB system was found to play an important role in maintaining and promoting the expression of PCNA, which will provide a theoretical basis for marker-assisted breeding of cashmere goats.

An important role of podoplanin in hair follicle growth.

Podoplanin (PDPN) is a glycoprotein that is expressed by various cell types, including keratinocytes, fibroblasts, and lymphatic endothelial cells. We found that PDPN is expressed in the hair follicle (HF) keratinocyte region and HF stem cell area during the late anagen phase but not during the telogen phase in mice. Importantly, keratinocyte-specific PDPN deletion in mice (K5-Cre;PDPNflox/flox) promoted anagen HF growth after depilation-induced HF regeneration as compared to control mice. RNA sequencing, followed by gene ontology analysis, showed down-regulation of focal adhesion and extracellular matrix interaction pathways in HF stem cells isolated from K5-Cre;PDPNflox/flox mice as compared to control mice. Furthermore, HF keratinocytes isolated from K5-Cre;PDPNflox/flox mice exhibited a decreased ability to interact with collagen type I in cell adhesion assays. Taken together, these results show that PDPN deletion promotes HF cycling, possibly via reduced focal adhesion and concomitantly enhanced migration of HF stem cells towards the bulb region. They also indicate potential new therapeutic strategies for the treatment of conditions associated with hair loss.

Hair Growth Promoting Effect of 4HGF Encapsulated with PGA Nanoparticles (PGA-4HGF) by β-Catenin Activation and Its Related Cell Cycle Molecules.

Poly-γ-glutamic acid (γ-PGA)-based nanoparticles draw remarkable attention as drug delivery agents due to their controlled release characteristics, low toxicity, and biocompatibility. 4HGF is an herbal mixture of Phellinus linteus grown on germinated brown rice, Cordyceps militaris grown on germinated soybeans, Polygonum multiflorum, Ficus carica, and Cocos nucifera oil. Here, we encapsulated 4HGF within PGA-based hydrogel nanoparticles, prepared by simple ionic gelation with chitosan, to facilitate its penetration into hair follicles (HFs). In this study, we report the hair promoting activity of 4HGF encapsulated with PGA nanoparticles (PGA-4HGF) and their mechanism, compared to 4HGF alone. The average size of spherical nanoparticles was ~400 nm in diameter. Continuous release of PGA-4HGF was observed in a simulated physiological condition. As expected, PGA-4HGF treatment increased hair length, induced earlier anagen initiation, and elongated the duration of the anagen phase in C57BL/6N mice, compared with free 4HGF treatment. PGA-4HGF significantly increased dermal papilla cell proliferation and induced cell cycle progression. PGA-4HGF also significantly increased the total amount of β-catenin protein expression, a stimulator of the anagen phase, through induction of cyclinD1 and CDK4 protein levels, compared to free 4HGF treatment. Our findings underscore the potential of PGA nanocapsules to efficiently deliver 4HGF into HFs, hence promoting hair-growth. Therefore, PGA-4HGF nanoparticles may be promising therapeutic agents for hair growth disorders.

Nanoindentation of Hair Cortex and Medulla Regions

Reports related to cuticle damage and hair’s mechanical properties are of interest to dermatologists in treatment of hair disorders. Apart from tensile test, few studies were reported on mechanical properties of hair, especially nanomechanical properties of cortex and medulla regions. Inner diameter of hair fiber (8–12 µm) is considered as medulla and it exists in hair at the beginning of the anagen phase. This study is aimed for quantitative comparison of nanomechanical properties between cortex and medulla and also with respect to donor age by performing nanoindentation on fiber cross sections using two different instruments. The medulla and cortex regions revealed hardness values of 70–170 MPa and 200–312 MPa, respectively. Similarly lower values of indentation modulus were observed for medulla compared to cortex because medulla has poorer orientation of microfibrils compared to the surrounding cortex. This study also found a linear correlation between hardness and indentation modulus especially for medulla region. This could be due to the fact that organization of the microfibrils increases from the medulla center to medulla periphery. It is also found that donor age has no influence on cortex modulus and hardness. The findings of this study will be useful for dermatologists in estimating the hair disorders and for estimating dye uptake of keratin fibers in textiles.

β-catenin-mediated hair growth induction effect of 3,4,5-tri-O-caffeoylquinic acid.

The hair follicle is a complex structure that goes through a cyclic period of growth (anagen), regression (catagen), and rest (telogen) under the regulation of several signaling pathways, including Wnt/ β-catenin, FGF, Shh, and Notch. The Wnt/β-catenin signaling is specifically involved in hair follicle morphogenesis, regeneration, and growth. β-catenin is expressed in the dermal papilla and promotes anagen induction and duration, as well as keratinocyte regulation and differentiation. In this study, we demonstrated the activation of β-catenin by a polyphenolic compound 3,4,5-tri-O-caffeoylquinic acid (TCQA) in mice model and in human dermal papilla cells to promote hair growth cycle. A complete regrowth of the shaved area of C3H mice was observed upon treatment with TCQA. Global gene expression analysis using microarray showed an upregulation in hair growth-associated genes. Moreover, the expression of β-catenin was remarkably upregulated in vivo and in vitro. These findings suggest that β-catenin activation by TCQA promoted the initiation of the anagen phase of the hair cycle.

Time course detection of dihydrocodeine in body hair after a single dose

BackgroundWhen head hair is not suitable or not available, body hair, such as leg or beard hair might be the most suitable sample for drug hair analysis. Information about the time course of drugs in hair, from the different anatomical body sites, should still be well documented. AimThe aim of this study was to determine and compare (a) the detection window of dihydrocodeine in frequently shaved legs and beard, (b) in unshaved hair from head hair, chest hair, leg hair, and/or arm hair, and (c) the distribution concentrations over the scalp, after a single dihydrocodeine intake. MethodBefore a single intake of 12 mg dihydrocodeine by subject 1 (woman), both legs hair were shaved in the morning. The subject 2 (man) shaved his beard in the morning and 30 min later he had a dose of 10 mg of dihydrocodeine.The samples were washed with water and shampoo, dried and collected as follows: Subject 1: every 3-days shaved leg hair (n = 9) and 1-month-later head hair (n = 1). Subject 2: daily shaved beard hair (n = 15), 2 months later head hair (n = 145), and every 20 days unshaved arm, leg and chest hair (from different areas) (n = 4/area). The samples were analysed for dihydrocodeine using a validated liquid chromatography-tandem mass spectrometry method with a limit of quantification (LOQ) of 15.6 pg/mg for dihydrocodeine.About 20 mg of hair samples were weighted, washed with dichloromethane, centrifuged, dried, and pulverized in the same disposable tubes. Then the samples were incubated with methanol (under sonication at 45 °C) during 4 h. After centrifugation, the supernatant was evaporated and a cation exchange solid phase extraction followed by separation and quantification using ultra performance liquid chromatography-tandem mass spectrometry (ULC-MS/MS) was carried out. Chromatographic separation was achieved using a BEH phenyl column eluted with 0.1% formic acid: methanol (0.1% formic acid). The UPLC-MS/MS method was validated and used in routine for drug hair analysis for already several years. Results and discussionIn the present study leg hair was collected every 3 days, as an average of frequent shaved hair in western woman population. Shaved leg hair was very limited and only one hair sample was available per analysis. Beard was collected daily and in a higher amount. Dihydrocodeine was detected in leg hair from the first sample (3 days after the intake). Maximum concentration at 68 pg/mg for the single intake was obtained after 15 days (±2 days), decreasing later to the LOQ from the 21th day. Beard hair was positive from the first day sample, and the maximum concentration was observed at 66 pg/mg, 6 days after the intake, decreasing later to the LOQ from day 13. This may be explained by growth rate and the amount of growing hairs, in anagen phase. In other body hair samples, dihydrocodeine was negative or detected from 1 month after the intake. No significant differences in dihydrocodeine concentrations over the scalp in the different regions were observed (p > 0.05). ConclusionBody hair presents different time course window detection due to the different growth rates. Frequently shaved leg and beard hair may be suitable samples for recent single dihydrocodeine dose detection from the first days up to 2–3 weeks after the intake, respectively, when a LOQ of 15.6 pg/mg is applied.

The Effect of a Combination of an Arginine Silicate Complex and Magnesium Biotinate on Hair and Nail Growth in Rats (P06-026-19)

ObjectivesAging, as well as inadequate intake of vitamins and minerals, can negatively affect the growth, strength, and appearance of hair and nails. For instance, biotin deficiency leads to brittle nails and hair loss. Moreover, certain nutrients, such as arginine and silicon, promote the biological conditions needed for optimal hair and nail growth. JDS-ASB is a novel combination of inositol-stabilized arginine silicate complex (ASI), a highly bioavailable form of arginine and silicon, and magnesium biotinate (MgB), a novel, well-absorbed biotin salt. The purpose of this study was to observe the effects of JDS-ASB on hair and nail growth in rats. MethodsTwenty-one female Sprague-Dawley rats (8-weeks old) were randomized into one of the following groups; 1: Control (no treatment), 2: ASI + Low Dose MgB (140 mg human equivalent dose (HED) of ASI and 1.5 mg HED of MgB), 3: ASI + High Dose MgB (140 mg HED of ASI and 10 mg HED of MgB). Study product was dosed daily by oral gavage for six weeks. Rat diets included egg whites to standardize absorbable biotin levels. At baseline, hair was shaved from the dorsal area of the rats and hair growth was assessed on days 7, 14, 21, 28, 35, and 42. Hair growth measurements included hair density (per square cm), and percentage of hair follicles in the anagen (growing phase) and telogen (resting phase) phases of the hair cycle. Mean nail growth was measured on days 14, 28, and 42. Blood samples were collected on day 42. ResultsVisual observations revealed greater hair growth in both treatment groups compared to control, however the effect was more pronounced in group 3. Compared to control, the percentage of hair follicles in the anagen phase was significantly greater in group 3 from day 14 onward (Figure 1). Compared to control, hair density was significantly greater in group 3 from day 21 onward. These parameters improved in group 2 compared to control on day 42. Blood levels of vascular endothelial growth factor, a protein associated with hair growth, were higher in both treatment groups compared to control, with levels greatest in group 3 (p < 0.05). Mean nail growth data showed faster daily growth in group 3 compared to other groups (p < 0.05). ConclusionsThe results of this study demonstrate that the combination of an arginine silicate complex and magnesium biotinate promotes hair and nail growth. Funding SourcesThis study was funded by JDS Therapeutics, LLC. Supporting Tables, Images and/or Graphs▪

Advances in Regenerative Stem Cell Therapy in Androgenic Alopecia and Hair Loss: Wnt pathway, Growth-Factor, and Mesenchymal Stem Cell Signaling Impact Analysis on Cell Growth and Hair Follicle Development.

The use of stem cells has been reported to improve hair regrowth in several therapeutic strategies, including reversing the pathological mechanisms, that contribute to hair loss, regeneration of hair follicles, or creating hair using the tissue-engineering approach. Although various promising stem cell approaches are progressing via pre-clinical models to clinical trials, intraoperative stem cell treatments with a one-step procedure offer a quicker result by incorporating an autologous cell source without manipulation, which may be injected by surgeons through a well-established clinical practice. Many authors have concentrated on adipose-derived stromal vascular cells due to their ability to separate into numerous cell genealogies, platelet-rich plasma for its ability to enhance cell multiplication and neo-angiogenesis, as well as human follicle mesenchymal stem cells. In this paper, the significant improvements in intraoperative stem cell approaches, from in vivo models to clinical investigations, are reviewed. The potential regenerative instruments and functions of various cell populaces in the hair regrowth process are discussed. The addition of Wnt signaling in dermal papilla cells is considered a key factor in stimulating hair growth. Mesenchymal stem cell-derived signaling and growth factors obtained by platelets influence hair growth through cellular proliferation to prolong the anagen phase (FGF-7), induce cell growth (ERK activation), stimulate hair follicle development (β-catenin), and suppress apoptotic cues (Bcl-2 release and Akt activation).

Pathomechanisms of immune-mediated alopecia.

The hair follicle (HF) is a complex mini-organ that constantly undergoes dynamic cycles of growth and regression throughout life. While proper progression of the hair cycle requires homeostatic interplay between the HF and its immune microenvironment, specific parts of the HF, such as the bulge throughout the hair cycle and the bulb in the anagen phase, maintain relative immune privilege (IP). When this IP collapses, inflammatory infiltrates that aggregate around the bulge and bulb launch an immune attack on the HF, resulting in hair loss or alopecia. Alopecia areata (AA) and primary cicatricial alopecia (PCA) are two common forms of immune-mediated alopecias, and recent advancements in understanding their disease mechanisms have accelerated the discovery of novel treatments for immune-mediated alopecias, specifically AA. In this review, we highlight the pathomechanisms involved in both AA and CA in hopes that a deeper understanding of their underlying disease pathogenesis will encourage the development of more effective treatments that can target distinct disease pathways with greater specificity while minimizing adverse effects.

Time course detection of dihydrocodeine in body hair after a single dose

When head hair is not suitable or not available, body hair, such as leg or beard hair might be the most suitable sample for drug hair analysis. Drugs may be detected in beard, from the first day up to more than one week. Specific information about the time course detection of drugs, after a single intake, in body hair has not been yet well documented for the different body hair samples. The aim of this study was to determine and compare the detection window of dihydrocodeine in frequently shaved legs and beard after a single intake. Unshaved hair from head hair, chest hair, leg hair, and/or arm hair were also analysed. Before a single intake of 12 mg dihydrocodeine by test subject 1 (female), the leg hair was shaved in the morning. The second test subject 2 (male) shaved his beard in the morning and 30 min later he had a dose of 10 mg of dihydrocodeine. The samples were washed with water and shampoo, dried and collected as follows: subject 1: every 3-days shaved leg hair ( n = 10) and 1-month-later head hair ( n = 1). Subject 2: daily shaved beard hair ( n = 15), 2 months later head hair ( n = 50, different regions), and every 20 days unshaved arm, leg and chest hair (from different regions) ( n = 4/area). The samples were analysed for dihydrocodeine using a validated liquid chromatography tandem mass spectrometry method with a limit of quantification (LOQ) of 16 pg/mg for dihydrocodeine. About 20 mg of hair samples were weighted, washed with dichloromethane, centrifuged, dried, and pulverized in the same disposable tubes. Then the samples were incubated with methanol (under sonication at 45 °C) during 4 h. After centrifugation, the supernatant was evaporated and a cation exchange solid phase extraction followed by separation and quantification using ultra performance liquid chromatography tandem mass spectrometry (UHLC-MS/MS) was carried out. Chromatographic separation was achieved using a BEH phenyl column eluted with 0.1% formic acid: methanol (0.1% formic acid). The UPLC-MS/MS method was validated and used in routine for drug hair analysis for already several years [1] . In the present study leg hair was collected every 3 days, as an average of frequent shaved hair in western woman population. Leg hair was very limited and only one hair sample from both woman legs was available per analysis. Beard was collected daily and in a higher amount. Dihydrocodeine was detected in leg hair from the first sample (3 days after the intake). Maximum concentration at 68 pg/mg for the single intake was obtained after 15 days (± 2 days), decreasing later to the LOQ from the 21st day. Beard hair was already positive from the first day sample, and the maximum concentration was observed at 66 pg/mg, 6 days after the intake, decreasing later to the LOQ from day 10. This may be explained by the ‘Richards–Meharg body and head hair growth table, where it indicates that legs have only 20% of growing hairs in anagen phase, while beard goes from 50 to 70%. In other body hair samples, dihydrocodeine was negative or detected from 1 month after the intake. Body hair presents different time course window detection due to the different growth rates. Frequently shaved leg and beard hair may be suitable samples for recent single dihydrocodeine dose detection from the first days up to 21 or 13 days after the intake, respectively, when an LOQ of 16 pg/mg is applied.

881 Hair growth and protection properties of an oligosaccharide and peptidic extract from Maca root

The hair follicle is a complex mini-organ that possesses the ability to self-renew through a specific growth cycle composed of 3 distinct phases: anagen (growth phase), catagen (regression phase) and telogen (hair elimination). During hair loss, in particular alopecia, the hair cycle is disrupted with a modification of the anagen/telogen ratio in favor of telogen phase. This results in an atrophy of the hair follicle thus producing shorter and thinner hair until it becomes microscopic. Maca (Lepidium meyenii) is a plant that grows at high altitudes in the Peruvian Andes. We have developed a patented active ingredient from Maca root, “Maca hydrolysate”. This extract, rich in oligosaccharides and peptides, is obtained from maca flour by a bioenzymatic process. Its biological efficacy has been evaluated in in vitro and ex vivo models on key factors involved in hair protection and hair growth. Maca extract was able to protect hair follicles from oxidative and inflammatory stress by decreasing the production of reactive oxygen species induced by lipid peroxide exposure in human follicle dermal papilla cells (HFDPC). The extract also decreased PGE2 release in HFDPC stressed by IL1-alpha, suggesting that Maca hydrolysate exerts anti-inflammatory effect. In a co-culture of human keratinocytes with HFDPC, the Maca extract significantly induced the production of FGF7 and beta–catenin, suggesting that it may induce the promotion of hair growth. Moreover, immunostainings of microdissected human hair follicles suggested that the extract could extend anagen phase and delay catagen. Accordingly, we have finally demonstrated that the extract significantly stimulated hair growth in ex vivo human hair follicles. Taken together, our data show that Maca hydrolysate extract is able to protect hair follicle and to promote hair growth, thereby slowing down hair loss. This new extract may be of particular interest in formulated products designed for the management of hair loss condition.

098 Simvastatin decreases AA-associated inflammation through effects on isoprenoid metabolites

Alopecia areata (AA) is a T-cell-mediated disorder in which lymphocytic infiltrate attacks and damages the hair follicle during the anagen phase of growth, leading to varying patterns of hair loss. At present, there is no treatment approved by the FDA for AA, and those treatments currently in use are associated with worrying side effects, prohibitive expense, or low rates of response. Previously, we have demonstrated the effectiveness of the lipid-lowering treatments simvastatin and ezetimibe in restoring T regulatory cell levels and hair growth in a mouse model of AA. Here, we aim to elucidate the mechanisms by which simvastatin exerts its effects on the autoimmune condition. Hair re-growth in the mouse model was associated with reduction of inflammatory markers in the skin, as well as reduced activation of STAT1 and reduced expression of NKG2D in skin-draining lymph nodes. These changes corresponded to a significant decrease in serum cholesterol and triglycerides in mice who responded with complete hair re-growth. Simvastatin was similarly able to decrease in vitro proliferation of both the IL-2-dependent cytotoxic T cell line CTLL-2 and primary mouse CD4+ and CD8+ T lymphocytes. This effect was shown to be at least partially mediated through the statin’s effects on isoprenoid metabolite production, as the addition of exogenous farnesyl pyrophosphate and geranyl geranyl pyrophosphate restored the proliferative ability of statin-treated CTLL-2 cells. Taken together with previous findings, these results indicate an immediate effect of simvastatin on the lymphocytic subpopulations responsible for hair follicle damage in AA through disruption of isoprenylation.

An overview of herbal alternatives in androgenetic alopecia.

The second most common alopecia-Androgenetic alopecia (AGA)-occurs due to hormonal imbalance. Dihydrotestosterone (DHT) an androgenic hormone is a sex steroid, produced in the gonads. The target sites of DHT are similar to that of testosterone, and it attaches easily remaining bound for 53minutes as compared to 35minutes of testosterone. Excess of DHT causes miniaturization of hair reducing the anagen phase and increasing the telogen phase leading to hair loss. Normally up to ten percent of testosterone in the body irreversibly gets converted into DHT by the action of enzyme 5-alpha-reductase. Inadequate blood flow to the scalp can also be another reason for hair loss encountered due to lower oxygen and nutrients reaching it. AGA affects both sexes; however in males, it leads to major hair loss. Conventional drugs such as minoxidil and finasteride are widely used for the treatment. However, several drawbacks such as allergic contact dermatitis, burning, ejaculation disorder, and decreased libido are reported. Available literature suggests the role of herbal drugs to have the action against 5-alpha-reductase enzyme inhibiting it and reducing the hair loss. This can be further potentiated since they exhibit lesser side effects. Recent advancements observed in the medicinal, cosmetic, and engineering fields can prove to be an asset. This article focuses on herbs which can be used in AGA. A review of Saw palmetto (Serenoa repens), Green tea (Camellia sinensis), Pumpkin seed (Curcurbita pepo), Rosemary (Rosmarinus officinalis), Grape seed (Vitis vinifera), and Licorice (Glycyrrhiza glabra) is attempted.