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Growth factors secreted by stem cells in dermis and subcutaneous adipose tissues and their roles in skin homeostasis
JSID Abstracts / Journal of Dermatological Science 69 (2013) e47–e93
e63
that of keratinocytes. It has previously been reported that administration of HSc025, a novel small compound that antagonizes the TGF-/Smad signal through the nuclear translocation of a transcriptional repressor YB-1, suppresses the development of dermal and hepatic fibrosis in mice. Here we examined the effects of HSc025 on cultured keratinocytes and fibroblasts in vitro, as well as on the incisional wound healing process in vivo. Treatment of primary cultures of human keratinocytes with HSc025 counteracted the inhibitory effects of TGF- on cell migration and proliferation in a dose-dependent manner. It also stimulated migration, but not proliferation, of primary human fibroblasts in vitro. After an incisional wounding created on the dorsal skin of mice, oral daily administration of HSc025 accelerated the wound closure from as early as Day 2. Experiments using transgenic type I collagen promoter/luciferase reporter mice indicated that, in contrast to its inhibitory effect on collagen expression in pathological fibrogenic condition, treatment of HSc025 did not affect collagen promoter activation during physiological wound healing process. These results lead to the better understanding of the cooperation of keratinocytes and fibroblasts in dermal wounding process and provide a novel insight into the therapy for impaired wound healing by using YB-1 modulators.
http://dx.doi.org/10.1016/j.jdermsci.2012.11.493
http://dx.doi.org/10.1016/j.jdermsci.2012.11.492
[Purpose] It has been reported that stem cells exist in skin tissues; however, the roles of stem cells are still not fully understood. We have specifically focused on stem cells in dermis and subcutaneous adipose tissues and studied on the potencies and functions of stem cells. In this study, focusing on growth factors secreted from stem cells, we examined the effects of these growth factors on homeostasis and regeneration of skin tissue. [Methods] First, growth factors expressed by human subcutaneous adipose-derived stem cells (hADSCs) were identified by microarray gene expression analysis. Secondly, using hairless mice (HR-1), growth factors were injected into the dorsal skin. And the changes in the skin were investigated by pathological evaluation and gene expression analysis. [Results and Discussion] The microarray analysis on hADSCs revealed gene expressions of several growth factors such as PDGF and bFGF. In the examination of effects of these growth factors on hADSCs and fibroblasts, it was found that the growth factors promoted proliferation of hADSCs and fibroblasts. When the growth factors were injected into dermis of mice, increased expressions of cell-proliferation related markers were confirmed compared to the control. There were also increased expressions of stem cell markers. Above results suggested that stem cell-secreted growth factors promote proliferation of stem cells and surrounding cells. In contrast, an opposite outcome was observed in the same study using old-age mice, which means cells in aged dermis might poorly respond to growth factors. Currently, we are conducting analysis of the agerelated changes of stem cells in the skin and examination of the ability for tissue regeneration.
P08-14 MicroRNA Expression Profiles in Keloid and Normal Dermal Fibroblasts Hajime Shimizu 1,∗ , Shinichi Igota 1,2 , Mamiko Egawa 1 , Mohammad Ghazizadeh 1
Tosa 3 , Seiko
1
Department of Molecular Pathology, Institute of Gerontology, Nippon Medical School 2 Department of Plastic and Reconstructive Surgery, Higashi-totsuka Memorial Hospital 3 Department of Plastic and Reconstructive Surgery, Nippon Medical School Musashi-kosugi Hospital Purpose: Keloids develop through abnormal wound healing process and are characterized by fibroblastic cell proliferation and excessive collagen synthesis but its etiology is still unknown. On the other hand, microRNAs are a highly conserved class of noncoding RNAs with important regulatory functions in cell proliferation, differentiation, and apoptosis. To discover novel regulatory pathways involved in keloid development, we performed microRNA expression profiling of keloid and normal dermal fibroblast cell cultures. Methods: Primary cultures of fibroblasts derived from keloid and normal dermis were used after being characterized in terms of cell proliferation and collagen synthesis. MicroRNA expression analysis was performed using microRNA microarray and the results were validated by a quantitative real-time PCR (q-PCR) method. Fibroblastic cell proliferation was assessed by a standard cell counting method and collagen synthesis was measured by an ELISA method.Results: MicroRNA expression analysis identified up-regulation (more than 2-fold) of miR-503, miR-886-3p, miR-129-3p, miR-199b-5p, miR-145, miR-19a, miR-221, miR-218, miR-7, miR-21 and miR-1274a and down-regulation (more than 2fold) of miR-10a and miR-1915. Validation by q-PCR confirmed the up-regulation of miR-19a, miR-503 and down-regulation of miR10a in keloid fibroblasts. Conclusion: Our data define the expression of micro-RNAs in keloid and normal dermal fibroblasts and suggest that miR-19a, miR-503 and miR-10a might form a significant regulatory network ultimately controlling the pathogenetic process of keloid formation. Further studies using microRNA inhibitors or activators will be needed to establish the exact role of these microRNA genes.
P08-15 Growth factors secreted by stem cells in dermis and subcutaneous adipose tissues and their roles in skin homeostasis Yuichi Hasebe 1,∗ , Shiro Ohgo 1 , Seiji Hasegawa 1,2 , Hiroshi Mizutani 1 , Satoru Nakata 1 , Akiko Yagami 2 , Kayoko Matsunaga 2 , Noriko Saitoh 3 , Kenji Matsumoto 3 , Masashi Toyoda 4 , Akihiro Umezawa 5 , Hirohiko Akamatsu 6 1
Reserch Laboratories Nippon Menard Cosmetic Co.,Ltd., Aichi, Japan Department of Dermatology, Fujita Health University School of Medicine, Aichi, Japan 3 Department of Allergy and Immunology, National Research Institute for Child Health and Development, Tokyo, Japan 4 Research Team for Geriatric Medicine (vascular medicine), Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan 5 Department of Reproductive Biology, National Research Institute for Child Health and Development, Tokyo, Japan 6 Department of Applied Cell and Regenerative Medicine, Fujita Health University Institute for Comprehensive Medical Science, Aichi, Japan 2
http://dx.doi.org/10.1016/j.jdermsci.2012.11.494
e63
that of keratinocytes. It has previously been reported that administration of HSc025, a novel small compound that antagonizes the TGF-/Smad signal through the nuclear translocation of a transcriptional repressor YB-1, suppresses the development of dermal and hepatic fibrosis in mice. Here we examined the effects of HSc025 on cultured keratinocytes and fibroblasts in vitro, as well as on the incisional wound healing process in vivo. Treatment of primary cultures of human keratinocytes with HSc025 counteracted the inhibitory effects of TGF- on cell migration and proliferation in a dose-dependent manner. It also stimulated migration, but not proliferation, of primary human fibroblasts in vitro. After an incisional wounding created on the dorsal skin of mice, oral daily administration of HSc025 accelerated the wound closure from as early as Day 2. Experiments using transgenic type I collagen promoter/luciferase reporter mice indicated that, in contrast to its inhibitory effect on collagen expression in pathological fibrogenic condition, treatment of HSc025 did not affect collagen promoter activation during physiological wound healing process. These results lead to the better understanding of the cooperation of keratinocytes and fibroblasts in dermal wounding process and provide a novel insight into the therapy for impaired wound healing by using YB-1 modulators.
http://dx.doi.org/10.1016/j.jdermsci.2012.11.493
http://dx.doi.org/10.1016/j.jdermsci.2012.11.492
[Purpose] It has been reported that stem cells exist in skin tissues; however, the roles of stem cells are still not fully understood. We have specifically focused on stem cells in dermis and subcutaneous adipose tissues and studied on the potencies and functions of stem cells. In this study, focusing on growth factors secreted from stem cells, we examined the effects of these growth factors on homeostasis and regeneration of skin tissue. [Methods] First, growth factors expressed by human subcutaneous adipose-derived stem cells (hADSCs) were identified by microarray gene expression analysis. Secondly, using hairless mice (HR-1), growth factors were injected into the dorsal skin. And the changes in the skin were investigated by pathological evaluation and gene expression analysis. [Results and Discussion] The microarray analysis on hADSCs revealed gene expressions of several growth factors such as PDGF and bFGF. In the examination of effects of these growth factors on hADSCs and fibroblasts, it was found that the growth factors promoted proliferation of hADSCs and fibroblasts. When the growth factors were injected into dermis of mice, increased expressions of cell-proliferation related markers were confirmed compared to the control. There were also increased expressions of stem cell markers. Above results suggested that stem cell-secreted growth factors promote proliferation of stem cells and surrounding cells. In contrast, an opposite outcome was observed in the same study using old-age mice, which means cells in aged dermis might poorly respond to growth factors. Currently, we are conducting analysis of the agerelated changes of stem cells in the skin and examination of the ability for tissue regeneration.
P08-14 MicroRNA Expression Profiles in Keloid and Normal Dermal Fibroblasts Hajime Shimizu 1,∗ , Shinichi Igota 1,2 , Mamiko Egawa 1 , Mohammad Ghazizadeh 1
Tosa 3 , Seiko
1
Department of Molecular Pathology, Institute of Gerontology, Nippon Medical School 2 Department of Plastic and Reconstructive Surgery, Higashi-totsuka Memorial Hospital 3 Department of Plastic and Reconstructive Surgery, Nippon Medical School Musashi-kosugi Hospital Purpose: Keloids develop through abnormal wound healing process and are characterized by fibroblastic cell proliferation and excessive collagen synthesis but its etiology is still unknown. On the other hand, microRNAs are a highly conserved class of noncoding RNAs with important regulatory functions in cell proliferation, differentiation, and apoptosis. To discover novel regulatory pathways involved in keloid development, we performed microRNA expression profiling of keloid and normal dermal fibroblast cell cultures. Methods: Primary cultures of fibroblasts derived from keloid and normal dermis were used after being characterized in terms of cell proliferation and collagen synthesis. MicroRNA expression analysis was performed using microRNA microarray and the results were validated by a quantitative real-time PCR (q-PCR) method. Fibroblastic cell proliferation was assessed by a standard cell counting method and collagen synthesis was measured by an ELISA method.Results: MicroRNA expression analysis identified up-regulation (more than 2-fold) of miR-503, miR-886-3p, miR-129-3p, miR-199b-5p, miR-145, miR-19a, miR-221, miR-218, miR-7, miR-21 and miR-1274a and down-regulation (more than 2fold) of miR-10a and miR-1915. Validation by q-PCR confirmed the up-regulation of miR-19a, miR-503 and down-regulation of miR10a in keloid fibroblasts. Conclusion: Our data define the expression of micro-RNAs in keloid and normal dermal fibroblasts and suggest that miR-19a, miR-503 and miR-10a might form a significant regulatory network ultimately controlling the pathogenetic process of keloid formation. Further studies using microRNA inhibitors or activators will be needed to establish the exact role of these microRNA genes.
P08-15 Growth factors secreted by stem cells in dermis and subcutaneous adipose tissues and their roles in skin homeostasis Yuichi Hasebe 1,∗ , Shiro Ohgo 1 , Seiji Hasegawa 1,2 , Hiroshi Mizutani 1 , Satoru Nakata 1 , Akiko Yagami 2 , Kayoko Matsunaga 2 , Noriko Saitoh 3 , Kenji Matsumoto 3 , Masashi Toyoda 4 , Akihiro Umezawa 5 , Hirohiko Akamatsu 6 1
Reserch Laboratories Nippon Menard Cosmetic Co.,Ltd., Aichi, Japan Department of Dermatology, Fujita Health University School of Medicine, Aichi, Japan 3 Department of Allergy and Immunology, National Research Institute for Child Health and Development, Tokyo, Japan 4 Research Team for Geriatric Medicine (vascular medicine), Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan 5 Department of Reproductive Biology, National Research Institute for Child Health and Development, Tokyo, Japan 6 Department of Applied Cell and Regenerative Medicine, Fujita Health University Institute for Comprehensive Medical Science, Aichi, Japan 2
http://dx.doi.org/10.1016/j.jdermsci.2012.11.494