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Hierarchy of cells and roles of stem cells in the dermis
Abstracts / Journal of Dermatological Science 84 (2016) e89–e180
skin care. However, its effects on epidermal keratinocytes remain largely unclear. In this study, human primary epidermal keratinocytes (HPEK) and human skin equivalent model were administrated with aloe vera gel, followed by the wound healing assay, flow cytometry analysis, and gene expression analysis. Our data indicated that aloe vera gel significantly improved wound healing. Moreover, flow cytometry analysis revealed that cell surface expression of 1-, ␣6-, 4-integrin, and E-cadherin were increased in HPEK treated with aloe vera gel, which may contribute to the cell migration and wound healing. Furthermore, treatment with aloe vera gel resulted in significant changes in cell-cycle progression and gene expression of differentiation markers in HPEK, suggesting roles of aloe vera in improvement of keratinocyte function. Therefore, HPEK were cultured with medium containing aloe vera gel at an air-liquid interface in order to construct human skin epidermal equivalents. The resultant equivalents are thicker and healthier than control equivalents, indicating the effectiveness of aloe vera on epidermal development as well. From these results, it is expected that aloe vera gel possesses a beneficial effect on wound healing and treatment for rough skin. http://dx.doi.org/10.1016/j.jdermsci.2016.08.450
e151
fibroblasts and stem cells). Using the dermis-derived cells, the proliferation potential and multipotency (adipocyte, osteoblast and chondrocyte) were examined. Further, gene expressions of each cell were compared to analyze overall condition of the cells in dermis. This study was approved by the Ethics Committee of Fujita Health University. Results and discussion: In the examination of multipotency of the dermis-derived cells obtained from fifty patients, stem cell-like cells which exhibit high multipotency were found in cells from several samples. Furthermore, gene expression analysis revealed specifically-high expressions of genes such as type-5 collagen in these cells. We continue further analysis to unravel other stem cell markers and the difference between the stem cell-like cells and fibroblasts and elucidate the mechanism underlying maintenance of homeostasis of the dermis. http://dx.doi.org/10.1016/j.jdermsci.2016.08.451 P08-20[O3-12] Role of topical or systemic corticosteroid application on hot water-induced mouse burn models of various grades
P08-19[O3-11]
Naotaka Doi ∗ , Yumi Nakatani, Yutaka Inaba, Nobuo Kanazawa, Fukumi Furukawa
Hierarchy of cells and roles of stem cells in the dermis
Dermatology, Wakayama Medical University, Wakayama, Japan
Yuichi Hasebe 1,2,∗ , Seiji Hasegawa 1,2,3 , Yuichirou Ogata 1 , Satoru Nakata 1 , Masashi Toyoda 4 , Kenji Matsumoto 5 , Akihiro Umezawa 6 , Kayoko Matsunaga 3 , Hirohiko Akamatsu 7
In clinical applications, corticosteroid ointment is generally used in the initial care for burn. Suppression of inflammation and subsequent pain and tissue damages is considered as the theoretical basis of this application, however, no clear evidence has ever been reported. The aim of this study is, using mouse models, to evaluate the effects of corticosteroid on burn and to clarify the underlying mechanisms, especially in its acute phase. With appropriate applications of hot water on back skin of adult male HR-1 mice, three-graded burn models (epidermal, dermal, subcutaneous burn) were established. To examine the effects of external corticosteroid application, diflorasone diacetate was painted on the back immediately after the generation of burn in some mice, while petrolatum was applied on control mice. To examine the effects of internal corticosteroid application, 1 mg/kg of prednisolone was orally applied immediately after the generation of burn in some mice. Then, lesional skin samples were obtained at various timepoints until 3 days after the generation of burn. Although histopathological and molecular analyses have not been completed, no obvious changes have been observed macroscopically with or without steroid treatment. On the other hand, corticosteroid applications seemed to decrease the sensation of sharp pain, which was evoked on the lesional skin with von-Frey filaments, in cases of the first and the third burn. The number of infiltrating inflammatory cells and expression level of proinflammatory cytokines in the lesional skin are now under investigation. It is expected that these results would provide some help for evaluation of the degree of recommendation on clinical corticosteroid application for burn therapy.
1
Nippon Menard Cosmetic Co., Ltd., Japan Nagoya University-MENARD Collaborative Research Chair, Nagoya University Graduate School of Medicine, Aichi, Japan 3 Department of Dermatology, Fujita Health University School of Medicine, Aichi, Japan 4 Research Team for Geriatric Medicine (vascular medicine), Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan 5 Department of Allergy and Immunology, National Research Institute for Child Health and Development, Tokyo, Japan 6 Department of Reproductive Biology, National Research Institute for Child Health and Development, Tokyo, Japan 7 Department of Applied Cell and Regenerative Medicine, Fujita Health University School of Medicine, Aichi, Japan 2
Purpose: It has been known that fibroblasts produce collagen fiber such as collagen, elastin and fibronectin in the dermis to maintain tissue homeostasis. However, the ecology of fibroblasts including their longevity and underlying mechanism of differentiation remains unknown. Recent studies have reported the existence of dermal stem cells, the origin of fibroblasts. We have also identified stem cells and several stem cell markers in the dermis. In this study, we investigated the characteristics of dermal stem cells by using the dermis of human skin and discussed the hierarchy of the cells that are involved in homeostasis of the dermis. Methods: In this study, we obtained normal dermis intraoperatively from fifty patients who underwent surgery after obtaining an informed consent. The dermal samples were reacted in collagenase to isolate dermis-derived cells (cell populations containing
http://dx.doi.org/10.1016/j.jdermsci.2016.08.452
skin care. However, its effects on epidermal keratinocytes remain largely unclear. In this study, human primary epidermal keratinocytes (HPEK) and human skin equivalent model were administrated with aloe vera gel, followed by the wound healing assay, flow cytometry analysis, and gene expression analysis. Our data indicated that aloe vera gel significantly improved wound healing. Moreover, flow cytometry analysis revealed that cell surface expression of 1-, ␣6-, 4-integrin, and E-cadherin were increased in HPEK treated with aloe vera gel, which may contribute to the cell migration and wound healing. Furthermore, treatment with aloe vera gel resulted in significant changes in cell-cycle progression and gene expression of differentiation markers in HPEK, suggesting roles of aloe vera in improvement of keratinocyte function. Therefore, HPEK were cultured with medium containing aloe vera gel at an air-liquid interface in order to construct human skin epidermal equivalents. The resultant equivalents are thicker and healthier than control equivalents, indicating the effectiveness of aloe vera on epidermal development as well. From these results, it is expected that aloe vera gel possesses a beneficial effect on wound healing and treatment for rough skin. http://dx.doi.org/10.1016/j.jdermsci.2016.08.450
e151
fibroblasts and stem cells). Using the dermis-derived cells, the proliferation potential and multipotency (adipocyte, osteoblast and chondrocyte) were examined. Further, gene expressions of each cell were compared to analyze overall condition of the cells in dermis. This study was approved by the Ethics Committee of Fujita Health University. Results and discussion: In the examination of multipotency of the dermis-derived cells obtained from fifty patients, stem cell-like cells which exhibit high multipotency were found in cells from several samples. Furthermore, gene expression analysis revealed specifically-high expressions of genes such as type-5 collagen in these cells. We continue further analysis to unravel other stem cell markers and the difference between the stem cell-like cells and fibroblasts and elucidate the mechanism underlying maintenance of homeostasis of the dermis. http://dx.doi.org/10.1016/j.jdermsci.2016.08.451 P08-20[O3-12] Role of topical or systemic corticosteroid application on hot water-induced mouse burn models of various grades
P08-19[O3-11]
Naotaka Doi ∗ , Yumi Nakatani, Yutaka Inaba, Nobuo Kanazawa, Fukumi Furukawa
Hierarchy of cells and roles of stem cells in the dermis
Dermatology, Wakayama Medical University, Wakayama, Japan
Yuichi Hasebe 1,2,∗ , Seiji Hasegawa 1,2,3 , Yuichirou Ogata 1 , Satoru Nakata 1 , Masashi Toyoda 4 , Kenji Matsumoto 5 , Akihiro Umezawa 6 , Kayoko Matsunaga 3 , Hirohiko Akamatsu 7
In clinical applications, corticosteroid ointment is generally used in the initial care for burn. Suppression of inflammation and subsequent pain and tissue damages is considered as the theoretical basis of this application, however, no clear evidence has ever been reported. The aim of this study is, using mouse models, to evaluate the effects of corticosteroid on burn and to clarify the underlying mechanisms, especially in its acute phase. With appropriate applications of hot water on back skin of adult male HR-1 mice, three-graded burn models (epidermal, dermal, subcutaneous burn) were established. To examine the effects of external corticosteroid application, diflorasone diacetate was painted on the back immediately after the generation of burn in some mice, while petrolatum was applied on control mice. To examine the effects of internal corticosteroid application, 1 mg/kg of prednisolone was orally applied immediately after the generation of burn in some mice. Then, lesional skin samples were obtained at various timepoints until 3 days after the generation of burn. Although histopathological and molecular analyses have not been completed, no obvious changes have been observed macroscopically with or without steroid treatment. On the other hand, corticosteroid applications seemed to decrease the sensation of sharp pain, which was evoked on the lesional skin with von-Frey filaments, in cases of the first and the third burn. The number of infiltrating inflammatory cells and expression level of proinflammatory cytokines in the lesional skin are now under investigation. It is expected that these results would provide some help for evaluation of the degree of recommendation on clinical corticosteroid application for burn therapy.
1
Nippon Menard Cosmetic Co., Ltd., Japan Nagoya University-MENARD Collaborative Research Chair, Nagoya University Graduate School of Medicine, Aichi, Japan 3 Department of Dermatology, Fujita Health University School of Medicine, Aichi, Japan 4 Research Team for Geriatric Medicine (vascular medicine), Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan 5 Department of Allergy and Immunology, National Research Institute for Child Health and Development, Tokyo, Japan 6 Department of Reproductive Biology, National Research Institute for Child Health and Development, Tokyo, Japan 7 Department of Applied Cell and Regenerative Medicine, Fujita Health University School of Medicine, Aichi, Japan 2
Purpose: It has been known that fibroblasts produce collagen fiber such as collagen, elastin and fibronectin in the dermis to maintain tissue homeostasis. However, the ecology of fibroblasts including their longevity and underlying mechanism of differentiation remains unknown. Recent studies have reported the existence of dermal stem cells, the origin of fibroblasts. We have also identified stem cells and several stem cell markers in the dermis. In this study, we investigated the characteristics of dermal stem cells by using the dermis of human skin and discussed the hierarchy of the cells that are involved in homeostasis of the dermis. Methods: In this study, we obtained normal dermis intraoperatively from fifty patients who underwent surgery after obtaining an informed consent. The dermal samples were reacted in collagenase to isolate dermis-derived cells (cell populations containing
http://dx.doi.org/10.1016/j.jdermsci.2016.08.452