PBT构建人工皮肤的试验研究.docxVIP

利用PEG/PBT构建人工皮肤的试验研究 作者：徐瑞兴 叶玲 杨华 陈双玲 杨丽 李幼忱 马俊杰 刘杰 【摘要】 目的 在聚乙二醇/聚对苯二甲酸丁二醇酯(PEG/PBT)嵌段共聚物中植入幼儿包皮成纤维细胞，构建人工真皮替代物，以研究细胞的生长、分化、增生及真表皮间的相互作用，为研制大面积创面覆盖物——复合皮奠定基础。方法 (1) 酶消化法培养幼儿成纤维细胞；(2)制备PEG/PBT膜，接种成纤维细胞后于不同时间观察细胞的形态变化；(3)H.E染色；(4)扫描电镜观测PEG/PBT膜的表面是否有成纤维细胞附着；(5)透射电镜观察膜上成纤维细胞形态及结构特点。结果 (1)利用PEG/PBT膜构建的人工真皮形成后质地与颜色近似真皮组织，有一定的弹性，可出现较明显的细胞增殖，未见人工真皮收缩现象。(2)H.E染色可见PEG/PBT膜呈较均一的淡红色网状分布，成纤维细胞主要分布于胶原膜表面。(3)扫描电镜下可见PEG/PBU膜表面有梭形的成纤维细胞出现，且成纤维细胞紧密贴附于PEG/PBT膜上。(4)透射电镜下见成纤维细胞功能活跃。结论 成纤维细胞在PEG/PBT膜中有着活跃的生物学特性，以PEG/PBT膜为支架构建的人工真皮具有同正常真皮相似的结构与功能。 【关键词】 成纤维细胞 PEG/PBT 皮肤组织工程 Study on how to make the artificial dermis by PEG/PBT membrane 【Abstract】 Objective To transplant the fibroblast as the seeding cell into the PEG/PBT membrane in order to investigate the growth， differentiation， increasing and the function between the epidermis and the dermis， simultaneously， prepare for making the artificial composite skin.Methods (1)Trypsin was used as the isolation medium to get the fibroblast；(2)Prepare for PEG/PBT membrane， the change of cell configuration was observed；(3)H.E staining；(4)Whether the cells attaching into the matrix was observed by scanning electron microscope(SEM)；(5)The construction features of fibroblast in the dermis were observed by transmission electron microscope(TEM).Results (1)The character and color of the artificial dermis were similar to the normal dermis， the artificial dermis was not fragile，proliferation of the cells could be clearly observed， and contraction of the dermis was not observed.(2)The PEG/PBT membrane was pale.red color by H.E staining， the fibroblasts were found on the surface of the matrix.(3)There were fusiform cells on the surface of the matrix by SEM， and the cells tightly attach into the membrane.(4)The fibroblasts in the membrane had active biologic function by transmission electron microscope.Conclusion The cultured fibroblasts in the membrane had active biologic character， the dermis made by PEG/PBT membra