| 用于替代皮肤刺激试验的组织工程表皮模型的构建研究 |
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| 引用本文： | 李中良,伍津津,杨桂红,奚廷斐,王春仁,陈亮.用于替代皮肤刺激试验的组织工程表皮模型的构建研究[J].中国修复重建外科杂志,2011(2):129-132. |
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| 作者姓名： | 李中良 伍津津 杨桂红 奚廷斐 王春仁 陈亮 |
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| 作者单位： | 温州医学院信息工程学院;第三军医大学大坪医院野战外科研究所皮肤科;中国药品生物制品检定所医疗器械中心; |
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| 基金项目： | 中国药品生物制品检定所资助项目(2009C3); 浙江省大学生科技创新活动计划(新苗人才计划)资助项目(2010R413057)~~ |
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| 摘 要： | 目的探讨人表皮细胞与聚碳酸脂膜构建组织工程表皮模型的方法,建立可用于皮肤刺激试验的组织工程表皮模型。方法应用组织工程方法,以聚碳酸脂膜为支架,以人皮肤角质形成细胞为细胞来源,构建组织工程表皮模型。气液面培养13 d,通过HE染色、角蛋白10(keratin 10,K10)和K13抗体、K14抗体、层粘连蛋白抗体、角化细胞交联外膜蛋白抗体、中间丝相关蛋白抗体免疫荧光染色、透射电镜观察组织工程表皮模型组织结构。采用快速渗透试验方法,SDS分别作用于培养基中添加脂质物(实验组)和未添加脂质物(对照组)的组织工程表皮模型18 h,测定组织活性减小50%所需化学物质的浓度(half maximal inhibitory concentration of a substance,IC50)值。结果 HE染色、免疫荧光染色及透射电镜观察显示,构建的组织工程表皮模型分化良好,具有与正常表皮相似的结构:基底膜、棘层、颗粒层和角化层。实验组和对照组组织工程表皮模型的IC50值分别为0.183%(6.00 mmol/L)和0.072%(2.36 mmol/L)。结论在聚碳酸酯膜上构建的组织工程表皮模型具有与正常表皮相似的组织结构,并且具有一定的屏障功能。
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| 关 键 词： | 组织工程表皮 皮肤刺激试验 替代试验 屏障功能 免疫荧光染色 |
EXPERIMENTAL STUDY ON CONSTRUCTING A TISSUE ENGINEERED EPIDERMIS FOR REPLACEMENT OF SKIN IRRITATION IN VITRO |
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| LI Zhongliang,WU Jinjin,YANG Guihong,XI Tingfei,WANG Chunren,CHEN Liang.Information & Engineering School of Wenzhou Medical College,Wenzhou Zhejiang,P.R.China.EXPERIMENTAL STUDY ON CONSTRUCTING A TISSUE ENGINEERED EPIDERMIS FOR REPLACEMENT OF SKIN IRRITATION IN VITRO[J].Chinese Journal of Reparative and Reconstructive Surgery,2011(2):129-132. |
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| Authors: | LI Zhongliang WU Jinjin YANG Guihong XI Tingfei WANG Chunren CHEN LiangInformation & Engineering School of Wenzhou Medical College Wenzhou Zhejiang PRChina |
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| Institution: | LI Zhongliang1,WU Jinjin2,YANG Guihong2,XI Tingfei3,WANG Chunren3,CHEN Liang3.1Information & Engineering School of Wenzhou Medical College,Wenzhou Zhejiang,325035,P.R.China,2Department of Dermatology,Institute of Surgery Research,Daping Hospital,Third Military Medical University,3Center of Medical Devices,National Institute for the Control of Pharmaceutical and Biological Products. |
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| Abstract: | Objective To investigate the method of constructing a tissue engineered epidermis with human epidermal cells and polycarbonate membrane,and to establish a tissue engineered epidermis with barrier function which is intended to be the replacing model in vitro of skin irritation test.Methods The tissue engineered epidermis was constructed by using polycarbonate membrane as scaffold and stratified differentiated epidermis derived from human keratinocytes.The tissue engineered epidermis was cultured on an inert ... |
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| Keywords: | Tissue engineered epidermis Skin irritation test Replacement test Barrier function Immunofluorescence staining |
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