以牛胶原凝胶为支架构建人工真皮的研究  

The study on making the artificial dermis from bovine collagen gel

作　　者：刘杰[1] 王德文[1] 崔雪梅[1] 张燕[1] 谷庆阳[1] 赵梅兰[1] 胡文华[1] 杨颖颙[2] 于淑贤[2] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]中国皮革和制鞋工业研究院,北京100016

出　　处：《军事医学科学院院刊》2004年第1期37-39,70,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：军事医学科学院创新基金 ( 990 5 10 6) ;放射医学研究所雏鹰计划资助项目 ;科技部科研院所技术开发专项资金项目 (NCSTE~ 2 0 0 0~JKZX~ 0 11)

摘　　要：目的 :制备成纤维细胞胶原凝胶 (即真皮替代物 ) ,以研究细胞的生长、分化、增生及与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酶消化法培养胎儿成纤维细胞 ;②制备细胞胶原凝胶 ,分为Ca_gel和Cf_gel组 ,接种后于不同时间观察两组细胞的形态变化和胶原凝胶的收缩情况 ;③细胞计数 ;④SEM观测胶原凝胶的表面是否有成纤维细胞附着 ;⑤HE染色。结果 :①成纤维细胞形态及增殖情况 :成纤维细胞胶原凝胶形成后呈半透明状 ,可出现较明显的细胞增殖 ,并可见凝胶收缩现象 ,但Ca_gel组较小 ;人工真皮质地与颜色近似真皮组织 ,有一定的弹性和抗拉性 ,不易脆。②细胞计数法所得两组生长曲线基本相似。Ca_gel和Cf_gel组细胞均具有典型的细胞增殖现象 ,Cf_gel组细胞在后期的增殖略快于Ca_gel组。③扫描显微镜下可见到凝胶表面有梭形的成纤维细胞出现 ,且成纤维细胞紧密贴附于凝胶上。④HE染色可见胶原呈较均一的淡红色网状分布 ,成纤维细胞分布于胶原凝胶内部和表面。结论 :①培养好的细胞凝胶称之为真皮替代物 ,是进行皮肤器官型培养和制作复合皮的关键与基础 ,并可与培养的人工表皮膜片混合移植 ,起到促进表皮细胞的生长黏附和加速创面愈合的作用。Objective:To transplant the fibroblasts as the seeding cells into the bovine collagen gel in order to investigate the growth, differentiation and proliferation of the cells and the interaction between the epidermis and the dermis, and to lay a foundation for preparing the artificial composite skin.Methods:①Trypsin was used to isolate the fibroblasts.②Fibroblast collagen gel was prepared. The artificial skin was divided into two groups,namely Ca-gel and Cf-gel.The changes in cell configuration and the shrinkage of the gel were also observed.③The cells were counted.④Whether the cells attached to the gel was observed under scanning electron microscope.⑤The cell collagen gel was stained with HE staining.Results:①Morphology and proliferation of fibroblasts: the fibroblast collagen gel was translucent, proliferation of the cells could be clearly observed, but after several days the gel contracted, the dermis of Ca-gel group less contracted than that of Cf-gel group.The character and color of the artificial dermis were similar to those of the normal dermis.The artificial dermis was not fragile.②The growth curve of the two groups was similar. Cell proliferation was found in both groups, but the cells of Cf-gel grew little faster than those of Ca-gel.③Fusiform cells on the surface of the gel were observed under scanning electronmicroscope, and the cells tightly stuck on the collagen gel.④The collagen gel was pale red in color after HE staining, the fibroblasts were found both on the surface and in the gel.Conclusions:①The cultured cell gel is named as dermis substitute and is the key point and base for making the artificial composite skin, and it could be transplanted to the wound with the cultured artificial epidermis. The keratinocytes can easily adhere to the dermis. The wound can heal quickly.②The fibroblasts in the collagen gel show actively biological character, so the dermis made by the collagen gel is an ideal substitute in research on the tissue engineering of skin or the study on th

关 键 词：成纤维细胞 胶原凝胶 人工真皮 组织工程 

分 类 号：R318.1[医药卫生—生物医学工程]