Stromal cells provide a crucial microenvironment for overlying epithelium. Here we investigated the expression and function of a stromal cell-specific protein, stromal cell-derived factor-1 （SDF-1）, in normal human ...Stromal cells provide a crucial microenvironment for overlying epithelium. Here we investigated the expression and function of a stromal cell-specific protein, stromal cell-derived factor-1 （SDF-1）, in normal human skin and in the tissues of diseased skin. Immunohistology and laser capture microdissection （LCM）-coupled quantitative real- time RT-PCR revealed that SDF-1 is constitutively and predominantly expressed in dermal stromal cells in nor- mal human skin in vivo. To our surprise, an extremely high level of SDF-1 transcription was observed in the dermis of normal human skin in vivo, evidenced by much higher mRNA expression level than type I collagen, the most abundant and highly expressed protein in human skin. SDF-1 was also upregulated in the tissues of many human skin disorders including psoriasis, basal cell carcinoma （BCC）, and squamous cell carcinoma （SCC）. Double immunostaining for SDF-1 and HSP47 （heat shock protein 47）, a marker of fibroblasts, revealed that fibroblasts were the major source of stroma-cell-derived SDF-1 in both normal and diseased skin. Functionally, SDF-1 activates the ERK （extracellular-signal-regulated kinases） pathway and functions as a mitogen to stimulate epidermalkeratinocyte proliferation. Both overexpression of SDF-1 in dermal fibroblasts and treatment with rhSDF-1 to the skin equivalent cultures significantly increased the number of keratinocyte layers and epidermal thickness. Con- versely, the stimulative function of SDF-1 on keratinocyte proliferation was nearly completely eliminated by inter- fering with CXCR4, a specific receptor of SDF-1, or by knock-down of SDF-1 in fibroblasts. Our data reveal that extremely high levels of SDF-1 provide a crucial microenvironment for epidermal keratinocyte proliferation in both physiologic and pathologic skin conditions.展开更多
目的可吸收生物材料聚羟乙酸(PGA)与成纤维细胞已被证实可以应用于体外组织工程肌腱的构建,本试验旨在探讨应用人皮肤成纤维细胞与PGA在体外进行组织构建时的适宜接种浓度。方法酶消化法获得人皮肤成纤维细胞经体外培养、扩增至第2代,...目的可吸收生物材料聚羟乙酸(PGA)与成纤维细胞已被证实可以应用于体外组织工程肌腱的构建,本试验旨在探讨应用人皮肤成纤维细胞与PGA在体外进行组织构建时的适宜接种浓度。方法酶消化法获得人皮肤成纤维细胞经体外培养、扩增至第2代,按照低、中、高三种浓度接种于PGA材料分别作为实验组1、2、3,传统浓度细胞接种的作为对照组,各组样本数n均为3,分别于接种后3天、1周、2周和4周进行观察和检测。结果接种后3天实验各组无明显差异,细胞在材料上黏附伸展良好。1周和2周时,各组细胞均有明显增殖,并分泌细胞外基质(ECM extra cellular matrix),但实验组1、2的ECM不如实验组3和对照组丰富,此后该现象更为明显。至第4周时,实验组1仅形成厚薄不均的薄膜状物,其余三组均形成组织样结构,但实验组2形成的细胞--材料复合物结构略为松散。结论应用本实验组中高浓度接种皮肤成纤维细胞在体外构建工程化组织可以达到传统所用更高浓度的接种效果,以节约细胞,减少取材。展开更多
Honey is widely used for treating burns, ulcers and wounds, but the mechanisms of action are poorly known and the product is mainlyused as an antimicrobial. We have examined here the wound healing properties of honey ...Honey is widely used for treating burns, ulcers and wounds, but the mechanisms of action are poorly known and the product is mainlyused as an antimicrobial. We have examined here the wound healing properties of honey on human fi broblasts, using an in vitroscratch wound healing model. Three kinds of widely used monofl oral honeys were used, viz. acacia (Robinia pseudacacia), buckwheat(Fagopyrum sp.), and manuka (Leptospermum scoparium). Data displayed an increased wound healing activity in fi broblasts, butwith diff erent effi ciency and mechanisms of action among honeys. The eff ects of acacia and buckwheat emerged in both scratchwound and chemotaxis assays, while the eff ect of manuka was signifi cant but lower. The use of inhibitors indicated on the wholean essential role of cytosolic calcium, an important role of ERK and p38, and a secondary role of PI3K. Acacia and buckwheat,but not manuka, induced signifi cant increases in the release of interleukin-4 (IL-4), IL-6, and IL-8, indicating a correlation betweeninterleukin upregulation and wound closure effi ciency. This is consistent with our previous fi ndings suggesting a higher ability ofacacia and buckwheat to activate keratinocyte reepithelialization, with respect to manuka honey. In conclusion, our data indicatethat acacia and buckwheat honeys are particularly effi cient in facilitating fi broblast wound closure activities, suggesting newtherapeutic possibilities for this natural product.展开更多
文摘Stromal cells provide a crucial microenvironment for overlying epithelium. Here we investigated the expression and function of a stromal cell-specific protein, stromal cell-derived factor-1 （SDF-1）, in normal human skin and in the tissues of diseased skin. Immunohistology and laser capture microdissection （LCM）-coupled quantitative real- time RT-PCR revealed that SDF-1 is constitutively and predominantly expressed in dermal stromal cells in nor- mal human skin in vivo. To our surprise, an extremely high level of SDF-1 transcription was observed in the dermis of normal human skin in vivo, evidenced by much higher mRNA expression level than type I collagen, the most abundant and highly expressed protein in human skin. SDF-1 was also upregulated in the tissues of many human skin disorders including psoriasis, basal cell carcinoma （BCC）, and squamous cell carcinoma （SCC）. Double immunostaining for SDF-1 and HSP47 （heat shock protein 47）, a marker of fibroblasts, revealed that fibroblasts were the major source of stroma-cell-derived SDF-1 in both normal and diseased skin. Functionally, SDF-1 activates the ERK （extracellular-signal-regulated kinases） pathway and functions as a mitogen to stimulate epidermalkeratinocyte proliferation. Both overexpression of SDF-1 in dermal fibroblasts and treatment with rhSDF-1 to the skin equivalent cultures significantly increased the number of keratinocyte layers and epidermal thickness. Con- versely, the stimulative function of SDF-1 on keratinocyte proliferation was nearly completely eliminated by inter- fering with CXCR4, a specific receptor of SDF-1, or by knock-down of SDF-1 in fibroblasts. Our data reveal that extremely high levels of SDF-1 provide a crucial microenvironment for epidermal keratinocyte proliferation in both physiologic and pathologic skin conditions.
文摘目的可吸收生物材料聚羟乙酸(PGA)与成纤维细胞已被证实可以应用于体外组织工程肌腱的构建,本试验旨在探讨应用人皮肤成纤维细胞与PGA在体外进行组织构建时的适宜接种浓度。方法酶消化法获得人皮肤成纤维细胞经体外培养、扩增至第2代,按照低、中、高三种浓度接种于PGA材料分别作为实验组1、2、3,传统浓度细胞接种的作为对照组,各组样本数n均为3,分别于接种后3天、1周、2周和4周进行观察和检测。结果接种后3天实验各组无明显差异,细胞在材料上黏附伸展良好。1周和2周时,各组细胞均有明显增殖,并分泌细胞外基质(ECM extra cellular matrix),但实验组1、2的ECM不如实验组3和对照组丰富,此后该现象更为明显。至第4周时,实验组1仅形成厚薄不均的薄膜状物,其余三组均形成组织样结构,但实验组2形成的细胞--材料复合物结构略为松散。结论应用本实验组中高浓度接种皮肤成纤维细胞在体外构建工程化组织可以达到传统所用更高浓度的接种效果,以节约细胞,减少取材。
基金Yamada Bee Farm Grant forHoneybee Research (2010).Yamada Bee Farm Grant for Honeybee Research
文摘Honey is widely used for treating burns, ulcers and wounds, but the mechanisms of action are poorly known and the product is mainlyused as an antimicrobial. We have examined here the wound healing properties of honey on human fi broblasts, using an in vitroscratch wound healing model. Three kinds of widely used monofl oral honeys were used, viz. acacia (Robinia pseudacacia), buckwheat(Fagopyrum sp.), and manuka (Leptospermum scoparium). Data displayed an increased wound healing activity in fi broblasts, butwith diff erent effi ciency and mechanisms of action among honeys. The eff ects of acacia and buckwheat emerged in both scratchwound and chemotaxis assays, while the eff ect of manuka was signifi cant but lower. The use of inhibitors indicated on the wholean essential role of cytosolic calcium, an important role of ERK and p38, and a secondary role of PI3K. Acacia and buckwheat,but not manuka, induced signifi cant increases in the release of interleukin-4 (IL-4), IL-6, and IL-8, indicating a correlation betweeninterleukin upregulation and wound closure effi ciency. This is consistent with our previous fi ndings suggesting a higher ability ofacacia and buckwheat to activate keratinocyte reepithelialization, with respect to manuka honey. In conclusion, our data indicatethat acacia and buckwheat honeys are particularly effi cient in facilitating fi broblast wound closure activities, suggesting newtherapeutic possibilities for this natural product.
