应用负载碱性成纤维细胞生长因子的人脱细胞羊膜构建人工活性真皮  

Construction of a new type of active skin substitutes for tissue engineering

作　　者：唐庆[1] 武日东[1] 唐诗[1] 林珣珣[1] 苏爱云[1] 

机构地区：[1]中山大学附属第一医院整形外科,广州510080

出　　处：《中华医学美学美容杂志》2012年第3期200-203,共4页Chinese Journal of Medical Aesthetics and Cosmetology

基　　金：广东省科技计划项目（编号：2008B030301336.2006B60501011）

摘　　要：目的探讨构建一种新型人工活性真皮的可行性。方法组织块法培养幼儿包皮成纤维细胞；采用酶一去垢剂法制备人脱细胞羊膜（HAAM）；双相法制备碱性成纤维细胞生长因子（bFGF）-明胶-壳聚糖缓释微球；缓释微球黏附于HAAM；bFGF基体式负载于HAAM．绘制药物缓释曲线；将第3～4代成纤维细胞培养于负载缓释微球的HAAM；扫描电镜观察HAAM、缓释微球的表而特征及缓释微球与HAAM的粘附情况；Western印迹法检测成纤维细胞中层粘连蛋白的表达。结果制备的HAAM为白色半透明状薄膜，有较高的孔隙率，空隙不规则。孔径大小为10～100nm，无细胞毒性；bFGF-明胶-壳聚精缓释微球分散较均匀，呈球形，粒径均匀．球体表面比较光滑，载药率为20ng／g，包封率为80．5％．体外药物缓释曲线最示药物控释效果良好；成纤维细胞在支架表面爬行生长良好，层粘连蛋白表达较对照组高。结论通过将成纤维细胞种植于负载bFGF-明胶-壳聚糖缓释微球的HAAM，有望制备出一种新型的人工活性真皮。Objective To discuss the possibility of constructing a new kind of active skin substi- tutes. Methods The culture of dermal fihroblasts were isolated from foreskin of an infant; prepara- tion of human acellular amniotic extracellular matrix was conducted through the disposition of deter- gent and enzyme. Surface structure porosity and pore size were detected through scanning electron mi- croscopy, bFGF chitosan-ge]atin microspheres were prepared. The features such as the size distribu- tion, drug content, drug encapsulating and in vitro release were studied with scanning electron micros- copy, laser grainsize analyzer and ELISA method. MTT test was performed to observe cell prolifera- tion and evaluate the biocompatihility in vitro. Results The cellular layer of amniotic membrane was completely removed hut did not damage the collogen scaffolds structure hy the disposal of enzyme and detergent. Scanning electron microscopy showed that the scaffolds had criss-cross structure and high porosity, the pore size was irregular and varied from 10 nm to 100 nm. Scanning electron microscopy of hFGF-gelatin-chitosan microspheres surface structure showed that the mierospheres had spherical structure, uniform size and smooth surface quality. Controlled release curve showed that the sudden release of bFGF was obvious. Conclusions A new type of active skin substitutes is prepared through culturing fibroblasts on HAAM loaded controlled released basic fibroblast growth factor from chi tosan-gelatin microspheres.

关 键 词：组织工程 支架 药物控释系统 

分 类 号：R318.1[医药卫生—生物医学工程]