• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.4
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• pp.674-682
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• 2011

The present study was undertaken to investigate the effect of acupuncture & moxibustion therapy on the hair follicle growth of skin 5 days and 10 days by macroscopic, microscopic and immunohistochemical methods. The results were as follows : Macroscopic hair follicle growth of plum-blossom needle treated group and strong moxibustion treated group was more increase than that of control group. Microscopic hair follicle growth of plum-blossom needle treated group and strong moxibustion treated group was hair growing cycle, anagen phase VI and that of control group and weak moxibustion treated group was hair growing cycle, anagen phase IV. Immunohistochemical observations on the expression of various growth factors, enzyme and receptor in hair follicle cycle after local treatment of acupuncture & moxibustion therapy are as follows: Expression of fibroblast growth factor was more intense in epidermis in plum-blossom needle treated group, epidermis and secondary hair germ cells in strong moxibustion treated group than control group. Expression of epidermal growth factor was more intense in epidermis in all experimental groups, and secondary hair germ cells in moxibustion treated group than control group. Expression of c-kit receptor was more intense in epidermis, secondary hair germ cells, outer root sheath in all experimental groups than control group. Expression of protein kinase C-${\alpha}$ was more intense in epidermis, secondary hair germ cells, outer root sheath in all experimental groups than control group. Expression of vascular endothelial growth factor was more intense in epidermis, bulge, secondary hair germ cells, outer root sheath in plum-blossom needle treated group and strong moxibustion treated group than control group. We concluded that acupuncture & moxibustion therapy related to the expression of various growth factors, enzymes and receptor on the hair growth cycle for hair growth.

• Biomolecules & Therapeutics
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• v.29 no.6
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• pp.643-649
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• 2021

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.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.113-117
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• 2003

The recent development of methods for culturing hair follicles in vitro has proved an important tool to investigate many aspects of drug screening. Hair follicles develop as a result of epithelial-mesenchymal interactions between epidermal keratinocytes and dermal cells. We isolated some follicle cells using explantation and enzymatic digestion method from human scalp hair follicles. So we could culture some follicular cells, such as outer root sheath (ORS) cells, dermal papilla (DP) cells, dermal sheath (DS) cells, matrix cells and melanocytes. To induce hair morphogenesis in vitro the cells were 3-D cultured as skin structures. Moreover, to develop hair follicel organ culture model, we applied dermal equivalent (DE) to culturing hair follicles to expand hair growth period.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.302-305
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• 2003

It is difficult to obtain sufficient healthy skin for coverage of a wide area of skin wound. In the skin, an additional population of living epithelial cells is located in the outer root sheath (ORS) of hair $follicles.^{1),2)}$ ORS cells should be a good source of epithelium because they are easily obtainable and patients do not have to suffer from scar formation at donor sites. We modified ordinary primary culture technique for the purpose of solving such problem that epithelial cells have a low propagation and easy aging during culture periods. First of all, we improved primary cultivation methods. In the ordinary primary culture, average yield of human ORS cells was $2\;{\times}\;10^3$ cells/follicle by direct incubation with trypsin (0.1%)/EDTA (0.02%) solution for 15 min at $37^{\circ}C$ but we could obtain about $6.5\;{\times}\;10^3$ cells/follicle by two step enzyme digestion method with dispase (1.2 U/ml) and trypsin (0.1%)/EDTA (0.02%) solution. So we could achieve three times higher primary cultured ORS cell yield. Secondly, we could obtain total $2\;{\times}\;10^7$ cells in serum free medium and even more total $6\;{\times}\;10^7$ cells in modified E-medium with mitomycin C-treated feeder cells during 17 days. Using the cultured ORS cells, and we could make bioartificial skin equivalent in vitro and concluded that ORS cells were progenitor cells for skin epithelial cell.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.1116-1124
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• 2009

To determine whether topical application of trimix extracts of Mylabris phalerata Pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata lead to affects on the hair growth activity in C57BL/6N mice. To examine the hair growth activity of the extracts of Mylabris phalerata Pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata gross, microscopic, and immunohistochemical method were performed. In order to examine the mRNA expression of hair growth related substance, RT-PCR method was performed. Experimental group I on day 14, The most extensive hair growth activity was observed in whole skin area of all the mice whose hair had been clipped. Brdu immunoreactive cells of all the experimental groups were more heavily stained in epidermis, bulge, outer root sheath, inner root sheath, subcutaneous tissue, hair bulb and cutaneous trunci muscles than that of control group on day 12 of hair growing cycle in C57BL/6N mice. VEGF immunoreactive density of all the experimental groups was more heavily stained in epidermis, bulge and cutaneous trunci muscles than that of control group on day 12. FGF and c-kit immunoreactive cells of all the experimental groups were heavily stained in epidermis, outer root sheath, inner root sheath and cutaneous trunci muscles on day 12. PKC-$\alpha$ immunoreactive density of all the experimental groups was mildly stained in epidermis and cutaneous trunci muscles than that of control group on day 12. On day 12, the expression of bFGF (138%, 119%, 120%), VEGF (146%, 144%, 133%), IGF-1 (165%, 141%, 119%) and PLI (121%, 116%, 123&) in each experimental groups was more increased than that of control group. On day 16, The expression of IGF-1 (126%, 149%, 151%) in all the experimental group was more increased than that of control group (100%). The expression of bFGF (92%, 94%) and VEGF (101%, 97%), PL1 (102%, 109%) in all the experimental group was more decreased than that of experimental group I, II on day 12. But the expression of bFGF (109%) and VEGF (127%), and PL1 (105%) in each experimental group III was more increased than that of control group (100%). These experiments suggest that trimix extracts of Mylabris phalerata pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata may stimulate the topical hair growth activity and its experimental group I can be useful for treatment of alopecia areata.

• Biomedical Science Letters
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• v.16 no.4
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• pp.349-357
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• 2010

We have examined the histological changes of skin during hair follicle growth after depilation in C57BL/6N mice. We first studied on histological changes of number of mast cells and thickness of skin during hair follicle growth periods (telogen, 1 day, 3 day, 5 day, 10 day, 14 day, 17 day and 21 day after depilation) by toluidine blue, Giemsa and H&E staining methods. We second studied immunoreactive density of cytokines and Brdu labeled cells in skin during hair follicle growth periods after depilation in C57BL/6N mice by immunohistochemical methods. The histological changes on skin thickness was increased from telogen to 14 day. The number of mast cells was decreased in 3,5 and 10 day and increased in 14, 17 and 20 day after depilation. Immunoreactive density of cytokines [protein kinase C-${\alpha}$ (PKC-${\alpha}$), c-kit, and vascular endothelial growth factor (VEGF)] in 1, 3, 5, 10, and 14 day after depilation was mildly stained in bulge and cutaneous trunci m., but immunoreactive density of cytokines in 17 and 21 day was heavily stained in epidermis, bulge, outer root sheath (ORS), inner root sheath (IRS) and cutaneous trunci m.. Immunoreactive density of Brdu labeled cells in skin in 1 and 3 day was heavily stained in bulge, epidermis and connective tissue under the cutaneous trunci m.. In all periods, immunoreactive density of Brdu labeled cells in skin was heavily stained in bulge, subcutaneous tissue, cutaneous trunci m, ORS and IRS. These experiments suggest that histological changes related to hair follicle growth elevated mast cell counts, skin thickness and epidermis thickness and heavily stained immunoreactive density of cytokines and Brdu labeled cutaneous trunci m. and connective tissue under the cutaneous trunci m. after depilation in C57BL/6N mice.

• Archives of Craniofacial Surgery
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• v.21 no.3
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• pp.176-179
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• 2020

Pilomatricoma is a benign tumor arising from the primitive basal cells of the epidermis that differentiate into hair matrix cells. Mutations in the CTNNB1 gene, which encodes β-catenin (a protein involved in hair growth), play an etiological role in the development of pilomatricoma. A 34-year-old woman presenting with a mass in the right parotid region underwent an excisional biopsy. The mass was conclusively diagnosed as pilomatricoma. During pregnancy, the mass grew from 1 cm to 5 cm in diameter and was accompanied by pain and tenderness. The growth may have been facilitated by the increased production of estrogen and progesterone, which bind to receptors located in the outer root sheath cells of the hair follicles. No recurrence was observed during 6 months of follow-up.

• International Journal of Oral Biology
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• v.36 no.3
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• pp.135-141
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• 2011

Hertwig's epithelial root sheath (HERS) consists of bi-layered cells derived from the inner and outer dental epithelia and plays important roles in tooth root formation as well as in the maintenance and regeneration of periodontal tissues. With regards to the fate of HERS, and although previous reports have suggested that this entails the formation of epithelial rests of Malassez, apoptosis or an epithelial-mesenchymal transformation (EMT), it is unclear what changes occur in the epithelial cells in this structure. This study examined whether HERS cells undergo EMT using a keratin-14 (K14) cre:ROSA 26 transgenic reporter mouse. The K14 transgene is expressed by many epithelial tissues, including the oral epithelium and the enamel organ. A distinct K14 expression pattern was found in the continuous HERS bi-layer and the epithelial diaphragm were visualized by detecting the ${\beta}$-galactosidase (lacZ) activity in 1 week postnatal mice. The 2 and 4 week old mice showed a fragmented HERS with cell aggregation along the root surface. However, some of the lacZ-positive dissociated cells along the root surface were not positive for pan-cytokeratin. These results suggest that the K14 transgene is a valuable marker of HERS. In addition, the current data suggest that some of the HERS cells may lose their epithelial properties after fragmentation and subsequently undergo EMT.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.151-157
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• 2003

Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.1
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• pp.75-85
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• 2023

This study was conducted to assess the effect of heptapeptide, composed of seven amino acids, on the activation of human hair cells isolated from human hair follicles. We have confirmed that the heptapeptide could bind to Lgr5 from the results of surface plasmon resonance (SPR) analysis. Heptapeptide enhanced the proliferation of human hair follicle dermal papilla cells (HHFDPCs) in a dose dependent manner. It induced the protein level of nuclear β-catenin, and the expressions of β-catenin downstream target genes, including LEF1, Cyc-D1 and c-Myc, in HHFDPCs. Heptapeptide significantly induced the phosphorylation of Akt and ERK, and the mRNA expressions of growth factors, including hepatocyte growth factor (HGF), keratinocyte growth factor (KGF) and vascular endothelial growth factor (VEGF), in HHFDPCs. In addition, heptapeptide significantly increased mRNA expression levels of differentiation-related transcription factors of human hair germinal matrix cells (HHGMCs) and differentiation markers of human hair outer root sheath cells (HHORSCs). Additionally, we investigated the effect of heptapeptide on human hair follicle stem cells (HHFSCs) differentiation and found that the heptapeptide reduced the mRNA and protein levels of stem cell markers, while it increased those levels of differentiation markers. These results have indicated that the heptapeptide promotes proliferation or differentiation of various types of hair follicle constituent cells through the induction of Wnt/β-catenin signaling. From the results, we have suggested that the heptapeptide in this study could be applied as a new functional material for the improvement of hair growth and alopecia.