Hair Growth Promoting Potential of Phospholipids Purified from Porcine Lung Tissues

BP201, porcine lung tissue-derived phospholipids, consists of phosphatidylcholine as a major phospholipid species. BP201 promoted hair growth after application onto the shaved backs of BALB/c and C3H mice. Its effect was enhanced when applied together with minoxidil (MNX) in C3H mice. When the tissue specimens prepared from the shaved skins of BP201-treated and control mice were microscopically examined, the total numbers of hair follicles in both anagen and telogen phases of BP201-treated mice were significantly higher than those of control mice. The numbers of hair follicles in the anagen phase of BP201-treated mice were also higher than those of control mice. In combination with MNX, BP201 further increased the total number of hair follicles, but did not alter the percentage of hair follicles in the anagenic phase. BP201 also increased the proliferation of human hair follicle dermal papilla cells. Collectively, BP201 possesses hair growth promoting potential, which would suggest its use singly or in combination for hair growth products.     

The additive effects of minoxidil and retinol on human hair growth in vitro

Minoxidil enhances hair growth by prolonging the anagen phase and induces new hair growth in androgenetic alopecia (AGA), whereas retinol significantly improves scalp skin condition and promotes hair growth. We investigated the combined effects of minoxidil and retinol on human hair growth in vitro and on cultured human dermal papilla cells (DPCs) and epidermal keratinocytes (HaCaT). The combination of minoxidil and retinol additively promoted hair growth in hair follicle organ cultures. In addition, minoxidil plus retinol more effectively elevated phosphorylated Erk, phosphorylated Akt levels, and the Bcl-2/Bax ratio than minoxidil alone in DPCs and HaCaT. We found that the significant hair shaft elongation demonstrated after minoxidil plus retinol treatment would depend on the dual kinetics associated with the activations of Erk- and Akt-dependent pathways and the prevention of apoptosis by increasing the Bcl-2/Bax ratio.     

Technologies for hair reconstruction and their applicability for pharmaceutical research

Hair follicles are the organs that produce hair shafts. They periodically regenerate throughout the life of the organisms, which is called the hair cycle. To develop new drugs to treat hair disorders and diseases, reproducible and high throughput assays or screening methods have been required to estimate the efficacy of various factors on hair follicle function. Although organ culture of hair follicles is one of the useful ways to carry out such research, it is not suitable for manipulating the genes or cells present in hair follicles. Patch assay is a method used to reconstruct hair follicles from enzymatically dissociated skin cells and has many advantages compared to the conventional Chamber method. Using the Patch method, transferring genes into follicular cells becomes easier than ever before. Chimeric follicles could be produced with dissociated cells by modifying the combination of cells or by simply merging cells of different origins. These applications certainly help the progress of hair research. However, we recently found that some functions of dermal papillae and follicular epithelia change during the growing phase (anagen) of the hair cycle. Dermal papillae produce different factors in early anagen and mid anagen. The signals from dermal papillae in early anagen could produce hair bulbs with clonogenic epithelial precursors but not with dormant epithelial precursors. On the other hand, the signals from dermal papillae in mid anagen strongly promote hair formation with dormant epithelial precursors. Therefore, more attention should be given to the hair cycle stages when using organ culture of hair follicles and conducting reconstruction experiments with follicularly derived cells.     

Natural scalp hair regression in preclinical stages of male androgenetic alopecia and its reversal by finasteride

Using contrast-enhanced phototrichogram (CE-PTG) at monthly intervals during 48 months, we measured the duration of the hair cycle, i.e. anagen, catagen and telogen at the exclusion of exogen. Exogen, a recently identified phase of the hair cycle, is characterized by weakening of anchorage of the club hair to the surrounding epithelium. The processing of the club hair terminates at the time of exogen hair release, i.e. hair shedding. We combined a noninvasive exogen sampling before each CE-PTG so that the area contained only anagen, catagen and telogen hair or empty follicular openings. During the first 24 months of this study, natural regression of hair cycling in early i.e. preclinical stages of androgenetic alopecia (AGA) in androgen sensitive areas was documented. Shortening of the hair cycle of thicker hair characterized progression of AGA. During the next 24 months, finasteride (1 mg/day) was introduced into the system. Shortening of the hair cycle was reversed by finasteride in androgen sensitive sites as long as the affected follicle was able to produce a thick hair fiber at the time of treatment initiation. Compared to the baseline period, responding follicles did not produce thicker hair. On average, they initiated active growth more rapidly by reducing the duration of the lag phase by 40%. The duration of the anagen phase of thick hair showed an average 23% increase. In this particular experiment, the already miniaturized follicles producing thinner hair (<40 microm thickness) at the time of finasteride introduction regressed further on treatment. Our results seem to indicate that reversal of 'hair loss' by finasteride probably means that the terminal type follicles that are functionally deficient--a stage of reversible hypotrophy--will be reactivated by two non-mutually exclusive mechanisms: faster regrowth followed by extension of the duration of anagen. In our study, there was no clear evidence in favour of reversal of miniaturized hair into terminal hair. This new interpretation indicates that miniaturized hair follicles may be an important diagnostic marker of AGA in males but also that it might be less contributive to the therapeutic response to finasteride. Our results highlight that precise measurement of terminal type hair follicle functionalities opens up avenues for the selection of 'drug-responsive organs' in the human scalp in vivo and these may possibly serve to predict 'quality of response to treatment'.     

Preparation of topical bimatoprost with enhanced skin infiltration and in vivo hair regrowth efficacy in androgenic alopecia

To prepare a topical formulation of bimatoprost (BIM) with high skin permeability, we designed a solvent mixture system composed of ethanol, diethylene glycol monoethyl ether, cyclomethicone, and butylated hydroxyanisole, serving as a volatile solvent, nonvolatile co-solvent, spreading agent, and antioxidant, respectively. The ideal topical BIM formulation (BIM–TF#5) exhibited 4.60-fold higher human skin flux and a 529% increase in dermal drug deposition compared to BIM in ethanol. In addition, compared to the other formulations, BIM–TF#5 maximally activated human dermal papilla cell proliferation at a concentration of 5 μM BIM, equivalent to 10 μM minoxidil. Moreover, BIM–TF#5 (0.3% [w/w] BIM) significantly promoted hair regrowth in the androgenic alopecia mouse model and increased the area covered by hair at 10 days by 585% compared to the vehicle-treated mice, indicating that entire telogen area transitioned into the anagen phase. Furthermore, at day 14, the hair weight of mice treated with BIM–TF#5 (5% [w/w] BIM) was 8.45- and 1.30-fold greater than in the 5% (w/w) BIM in ethanol and 5% (w/v) minoxidil treated groups, respectively. In the histological examination, the number and diameter of hair follicles in the deep subcutis were significantly increased in the BIM–TF#5 (0.3 or 5% [w/w] BIM)-treated mice compared to the mice treated with vehicle or 5% (w/w) BIM in ethanol. Thus, our findings suggest that BIM–TF#5 is an effective formulation to treat scalp alopecia, as part of a novel therapeutic approach involving direct prostamide F2α receptor-mediated stimulation of dermal papilla cells within hair follicles.     

Eclipta prostrata promotes the induction of anagen,sustains the anagen phase through regulation of FGF-7 and FGF-5

Context: Eclipta prostrata L. (Asteraceae) (EP) has been widely used for the treatment of skin disease in Asian traditional medicine.

Objective: This study investigates the potency of EP in promoting hair growth in vivo and in vitro.

Materials and methods: C57BL/6N mice were divided into four groups (n?=?4) as follows: control (topical treatment of normal saline), topical 3% minoxidil to the dorsal skin of mice for 14 days, and low (1?mg/day) and high (10?mg/day) doses of EP orally administered once a day for 14 days. Dorsal hairs of C57BL/6N mice were depilated to synchronize anagen induction. Hair growth activity was evaluated by gross and microscopic observations. Sections of dorsal skin were stained with haematoxylin and eosin. We also treated the various concentrations of EP (5, 10 and 50?μg/mL) for 24?h on the human dermal papilla cells (HDPs) and examined the effects of EP on the expression of FGF-7 and mTOR signalling.

Results: EP enhanced the induction of anagen in the dorsal skin of mice, characterized by the appearance of inner root sheath along with hair shaft, the emergence of hair shaft through the epidermis. EP increased the expression of FGF-7, while decreased the level of FGF-5 in C57/BL6 mice. EP also increased the expression of FGF-7, activated the mTOR signalling in HDPs.

Discussion and conclusions: These results suggest that EP has a potency to enhance the growth of hair follicle, promoting hair growth through regulation of FGF-7 and FGF-5.     

Squarticles as a Lipid Nanocarrier for Delivering Diphencyprone and Minoxidil to Hair Follicles and Human Dermal Papilla Cells

Delivery of diphencyprone (DPCP) and minoxidil to hair follicles and related cells is important in the treatment of alopecia. Here we report the development of “squarticles,” nanoparticles formed from sebum-derived lipids such as squalene and fatty esters, for use in achieving targeted drug delivery to the follicles. Two different nanosystems, nanostructured lipid carriers (NLC) and nanoemulsions (NE), were prepared. The physicochemical properties of squarticles, including size, zeta potential, drug encapsulation efficiency, and drug release, were examined. Squarticles were compared to a free control solution with respect to skin absorption, follicular accumulation, and dermal papilla cell targeting. The particle size of the NLC type was 177 nm; that of the NE type was 194 nm. Approximately 80% of DPCP and 60% of minoxidil were entrapped into squarticles. An improved drug deposition in the skin was observed in the in vitro absorption test. Compared to the free control, the squarticles reduced minoxidil penetration through the skin. This may indicate a minimized absorption into systemic circulation. Follicular uptake by squarticles was 2- and 7-fold higher for DPCP and minoxidil respectively compared to the free control. Fluorescence and confocal images of the skin confirmed a great accumulation of squarticles in the follicles and the deeper skin strata. Vascular endothelial growth factor expression in dermal papilla cells was significantly upregulated after the loading of minoxidil into the squarticles. In vitro papilla cell viability and in vivo skin irritancy tests in nude mice suggested a good tolerability of squarticles to skin. Squarticles provide a promising nanocarrier for topical delivery of DPCP and minoxidil.     

Hair growth regulation by the extract of aromatic plant <Emphasis Type="Italic">Erica multiflora</Emphasis>

Hair growth problems can affect human physical and mental health and are of particular relevance during the aging process. In an effort to resolve such problems, we attempted to find plants having hair growth regulation activity and thus collected plant extracts from Tunisia for bioprospecting purposes. Among them, we investigated the Erica multiflora extract to evaluate the hair growth promotion activity by using the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay and cell cycle assay on human dermal papilla cells in vitro and an administration assay on mouse dorsal skin in vivo. The results showed that the Erica multiflora extract promotes dermal papilla cell growth and cell cycle with high activity, and induced hair growth in vivo by induction of anagen phase from telogen phase.     

Promotion effect of acankoreoside J, a lupane-triterpene in Acanthopanax koreanum, on hair growth

This study was conducted to evaluate the effect of Acanthopanax koreanum and acankoreoside J from A. koreanum on the promotion of hair growth. When immortalized rat vibrissa dermal papilla cells were treated with extract of A. koreanum leaves, the proliferation of dermal papilla cells significantly increased. In particular, acankoreoside J among several components, isolated from A. koreanum leaves, markedly promoted the proliferation of the dermal papilla cells. When rat vibrissa follicles were treated with an acankoreoside J, the hair-fiber lengths of the vibrissa follicles increased significantly. We further investigated β-catenin pathway and cell cycle regulation with respect to the effect of acankoreoside J on the proliferation of the dermal papilla cells. Treatment with acankoreoside J results in an increase of nuclear β-catenin level, and up-regulation of cyclin D1, cyclin E and CDK2, whereas, the expression of p27(kip1) was down-regulated in the dermal papilla cells. Taken together, these results suggest that acankoreoside J, a lupane-triterpene of A. koreanum, has the potential of promoting hair growth by promoting cell cycle progression of the dermal papilla cells, through the increase of nuclear β-catenin, along with the up-regulation of cyclin D1, cyclin E and CDK2, and down-regulation of p27(kip1).     

Baicalin,a flavonoid,affects the activity of human dermal papilla cells and promotes anagen induction in mice

Baicalin, a flavonoid isolated from Scutellaria baicalensis, is known to have multiple biological functions. Recent studies have demonstrated that baicalin treatment increases alkaline phosphatase activity (ALP) and osteoprotegerin secretion by osteoblasts. Furthermore, baicalin induces the differentiation of cultured osteoblasts via the activation of the Wnt/β-catenin signaling pathway. In this study, we evaluated the hair growth-promoting effects of baicalin in human follicular dermal papilla (DP) cells. A reporter assay and Western blotting were used to assess the effect of baicalin on β-catenin signaling in DP cells. ALP activity and messenger RNA (mRNA) expression were examined by ALP staining and real-time polymerase chain reaction (PCR), respectively. Growth factor expression levels were also evaluated using real-time PCR. Finally, the effect of baicalin on hair growth in vivo was examined by topical application of baicalin on the shaved dorsal skin of C57BL/6 mice. Our results indicate that baicalin activates Wnt/β-catenin signaling in a dose-dependent manner in human DP cells. ALP mRNA expression and activity were significantly induced in the presence of baicalin. In addition, treatment with baicalin induced the mRNA expression of growth factors, such as insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF). Moreover, compared to vehicle treatment, baicalin treatment induced an earlier conversion from telogen to anagen. Our results strongly suggest that baicalin promotes hair growth by regulating the activity of DP cells.

     

The effect of tripeptide-copper complex on human hair growth in vitro

The tripeptide-copper complex, described as a growth factor for various kinds of differentiated cells, stimulates the proliferation of dermal fibroblasts and elevates the production of vascular endothelial growth factor, but decreased the secretion of transforming growth factor-beta1 by dermal fibroblasts. Dermal papilla cells (DPCs) are specialized fibroblasts, which are important in the morphogenesis and growth of hair follicles. In the present study, the effects of L-alanyl-L-histidyl-L-lysine-Cu2+ (AHK-Cu) on human hair growth ex vivo and cultured dermal papilla cells were evaluated. AHK-Cu (10(-12) - 10(-9) M) stimulated the elongation of human hair follicles ex vivo and the proliferation of DPCs in vitro. Annexin V-fluorescein isothiocyanate/propidium iodide labeling and flow cytometric analysis showed that 10(-9) M AHK-Cu reduced the number of apoptotic DPCs, but this decrease was not statistically significant. The ratio of Bcl-2/Bax was elevated, and the levels of the cleaved forms of caspase-3 and PARP were reduced by treatment with 10(-9) M AHK-Cu. The present study proposed that AHK-Cu promotes the growth of human hair follicles, and this stimulatory effect may occur due to stimulation of the proliferation and the preclusion of the apoptosis of DPCs.     

Preventive effects of cedrol against alopecia in cyclophosphamide-treated mice

Although numerous hypotheses have been proposed to prevent chemotherapy-induced alopecia (CIA), effective pharmaceuticals have yet to be developed. In our study, the back hairs of C57BL/6 mice were factitiously removed. These mice were then treated with cedrol or minoxidil daily. Mice with early-stage anagen VI hair follicles were treated with cyclophosphamide (CYP, 125 mg/kg) to induce alopecia. The CYP-damaged hair follicles were observed and quantified by using a digital photomicrograph. The results demonstrated that the minoxidil-treated mice suffered from complete alopecia similar to the model 6 days after CYP administration. Simultaneously, the cedrol-treated (200 mg/kg) mice manifested mild alopecia with 40% suppression. Histological observation revealed that anagen hair follicles of the cedrol-pretreated mice (82.5%) likely provided from damage compared with the sparse and dystrophic hair follicles of the model mice (37.0%). Therefore, the use of topical cedrol can prevent hair follicle dystrophy and provide local protection against CIA.     

The hair follicle and its stem cells as drug delivery targets

The hair follicle is a skin appendage with a complex structure containing many cell types that produce highly specialised proteins. The hair follicle is in a continuous cycle: anagen is the hair growth phase, catagen the involution phase and telogen is the resting phase. The follicle offers many potential therapeutic targets. Hoffman and colleagues have pioneered hair-follicle-specific targeting using liposomes to deliver small and large molecules, including genes. They have also pioneered ex vivo hair-follicle targeting with continued expression of the introduced gene following transplantation. Recently, it has been discovered that hair follicle stem cells are highly pluripotent and can form neurons, glial cells and other cell types, and this has suggested that hair follicle stem cells may serve as gene therapy targets for regenerative medicine.     

Drug-induced hair disorders

Drugs may induce hair loss, stimulate hair growth or, more rarely, induce changes in the hair shape and colour. Drug-induced hair loss is usually completely reversible and is, in most cases, a consequence of a toxic effect of the drug on the hair follicle matrix. In rare cases alopecia may be permanent. Depending on type of drug, dosage and patient susceptibility, hair loss presents as telogen effluvium, anagen effluvium or both. Telogen effluvium is also commonly observed after discontinuation of drugs that prolong anagen, such as topical minoxidil and oral contraceptives. Although a large number of drugs have been occasionally reported to produce hair loss, only for a few drugs the relation between drug intake and hair loss has been proven.     

Influence of hepatocyte growth factor/scatter factor (HGF/SF) on fibroblast growth factor-2 (FGF-2) levels in external auditory canal cholesteatoma (EACC) cell culture

BACKGROUND: In previous studies, we cited circulatory disorders and hypoxia as etiological factors for the formation of external auditory canal cholesteatoma (EACC) resulting in angiogenesis. Here, we investigate how the angiogenic factor hepatocyte growth factor/scatter factor (HGF/SF) influences the level of another angiogenic factor FGF-2. MATERIALS AND METHODS: After 16 to 72 hours of incubation with 20 ng/ml HGF/SF, levels of VEGF in the HGF/SF-treated and untreated culture was analyzed. We also investigated the influence of HGF/SF (20-80 ng/ml) on the concentration of FGF-2. RESULTS: After 16 hours of incubation with HGF/SF at 20 ng/ml, FGF-2 was measured at 44.19 pg/ml (control: 42.24 pg/ml). After 72 hours, FGF-2 was at 23.41 pg/ml (control: 14.83 pg/ml). After 24 hours, 40 ng/ml HGF/SF showed the highest concentration of FGF-2. CONCLUSION: FGF-2 levels were initially elevated after treatment with HGF/SF, however, further incubation did not show any increase. We assume that HGF/SF released FGF-2 in the matrix but did not induce FGF-2 expression. The most effective HGF/SF concentration was 40 ng/ml.     

Characterization of two novel forms of the rat sulphonylurea receptor SUR1A2 and SUR1BDelta31

1. The ATP-sensitive potassium channel (K(ATP)) of pancreatic beta-cells is composed of the sulphonylurea-binding protein, SUR1, and the inwardly rectifying K(+) channel subunit, Kir6.2. We have characterized two novel isoforms of rat SUR1 in the RINm5F insulin-secreting cell line. 2. SUR1A2 is an allelic variant with a single amino acid change in the first nucleotide-binding domain. Coinjection of SUR1A2 plus Kir6.2 into Xenopus oocytes or expression of a SUR1A2-Kir6.2 tandem in HEK-293 cells yielded large currents with characteristics similar to the wild-type K(ATP) channel. 3. SUR1BDelta31, detected in several human tissues, is a splice variant of the rat SUR1 gene that lacks exon 31 of the corresponding human SUR1 gene. SUR1BDelta31 lacks the TM16-TM17 transmembrane-spanning helices leading to a protein with a different transmembrane topology. Coinjection of SUR1BDelta31 plus Kir6.2 into Xenopus oocytes or expression of the Kir6.2/SUR1BDelta31 tandem construct in HEK-293 cells did not result in any current, and a surface expression assay indicated that this channel does not reach the plasma membrane. 4. SUR1A2 and SUR1A1 proteins expressed in HEK-293 cells display similar binding affinities for [(3)H]-glibenclamide, while SUR1BDelta31 shows a 500-fold lower affinity. 5. These findings confirm that TM16-TM17 of SUR1 are important for high-affinity glibenclamide binding and that their deletion impairs trafficking of the K(ATP) channel to the surface membrane.     

Cardioselectivity of the sulphonylurea HMR 1098: studies on native and recombinant cardiac and pancreatic K(ATP) channels

In this study we investigated the effects of the putative cardioselective sulphonylurea derivative HMR 1098 on ATP-sensitive potassium (K(ATP)) channels from cardiac ventricular myocytes, the INS-1 beta-cell line and from recombinant K(ATP) channels composed of SUR2A/Kir6.2, SUR1/Kir6.2, SUR1/Kir6.1 an Kir6.2,DeltaC26. Recombinant channels were expressed in tsA201 or COS-1 cells. The effects of HMR 1098 on single channel and whole-cell currents were recorded using the patch-clamp technique. At the single channel level, using excised inside-out membrane patches, HMR 1098 inhibited K(ATP) channels from ventricular cells and INS-1 cells with IC(50)s of 0.88 and 720 microM respectively. Similar results to those in cardiac cells were obtained using recombinant SUR2A/Kir6.2 K(ATP) channels. HMR 1098 inhibition of SUR2A/Kir6.2 K(ATP) channels was unaffected by the presence of internal ADP. In whole-cell recordings, HMR 1098 inhibited SUR2A/Kir6.2 and SUR1/Kir6.2 currents with IC(50)s of 2.1 and 860 microM respectively. HMR 1098 was without effect on currents either from the Kir6.2,DeltaC26 truncation mutant or from Kir2.1. Our results demonstrate that HMR 1098 is a selective inhibitor of cardiac K(ATP) channels, showing a 400 - 800-fold selectivity over beta-cell K(ATP) channels. The non-aromatic substitutions in the sulphonylurea moiety greatly increase the cardioselectivity of this compound while reducing the overall blocking potency of this sulphonylurea derivative.     

Hair Growth Promotion by δ-Opioid Receptor Activation

Literature has revealed that the delta opioid receptor (DOR) exhibited diverse pharmacological effects on neuron and skin. In the present study, we have investigated whether the activation of DOR has hair-growth promotion effects. Compared with other opioid receptor, DOR was highly expressed in epidermal component of hair follicle in human and rodents. The expression of DOR was high in the anagen phase, but it was low in the catagen and telogen phases during mouse hair cycle. Topical application of UFP-512, a specific DOR agonist, significantly accelerated the induction of the anagen in C₃H mice. Topical application of UFP-512 also increased the hair length in hair organ cultures and promoted the proliferation and the migration of outer root sheath (ORS) cells. Similarly, pharmacological inhibition of DOR by naltrindole significantly inhibited the anagen transition process and decreased hair length in hair organ cultures. Thus, we further examined whether Wnt/β-catenin pathway was related to the effects of DOR on hair growth. We found that Wnt/β-catenin pathway was activated by UFP-512 and siRNA for β-catenin attenuated the UFP-512 induced proliferation and migration of ORS cells. Collectively, result established that DOR was involved in hair cycle regulation, and that DOR agonists such as UFP-512 should be developed for novel hair-loss treatment.