选择性去细胞异种皮的制备及其生物相容性研究  

PREPARATION OF SELECTIVELY DECELLULAR XENOSKIN AND ITS BIOCOMPATIBILITY

作　　者：孙天骏[1] 柴家科[1] 韩焱福[1] 杨红明[1] 刘玲英[1] 苏鹏[1] 冯瑞[1] 

机构地区：[1]解放军总医院第一附属医院烧伤整形科,北京100048

出　　处：《中国修复重建外科杂志》2010年第6期668-672,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：卫生公益性行业科研专项资助项目(200802066);解放军总医院苗圃基金项目(08MP14)~~

摘　　要：目的探讨一种保留表皮层结构、真皮层去细胞、具有低抗原性的选择性去细胞异种皮制备方法 ,并研究其生物相容性。方法取健康白色家猪全厚皮,采用戊二醛-胰蛋白酶-去污剂的方法制备选择性去细胞异种皮。细胞毒性实验按照GB/T16886.5-2003《医疗器械生物学评价》体外细胞毒性实验方法进行,毒性评估标准参见美国药典。皮下植入实验按照GB/T16886.6-1997《医疗器械生物学评价》植入后局部反应实验方法进行,取成年Wistar大鼠72只,随机分为选择性去细胞异种皮、新鲜猪皮、SD大鼠异体皮3组(A、B、C组,n=24),于实验动物背部皮下分别植入1cm×1cm相应皮片,于术后1、2、4、8、12、16周行大体观察和组织学观察。结果细胞毒性实验显示选择性去细胞异种皮毒性判定为1级,符合国家医疗器械生物学评价标准。皮下植入实验显示,3组大鼠皮下均出现炎性反应,A组选择性去细胞异种皮和C组SD大鼠异体皮炎性反应较弱,真皮层内逐渐有新生血管生成和纤维样组织长入,植入皮片均逐渐降解;B组新鲜猪皮炎性反应较强,持续时间长,植入皮片逐渐变硬,不易降解。结论采用戊二醛-胰蛋白酶-去污剂制备的选择性去细胞异种皮具有双层皮肤结构的形态学优势和良好生物相容性,可作为生物材料修复大面积深度烧伤创面。Objective To prepare and study the biocompatibility of selectively decellular xenoskin which has the character of the lower antigen,continuous epidermis,and the dermal matrix without any cellular components.Methods The porcine skin was treated with glutaraldehyde solution,trypsin,and detergent solution TritonX-100 to prepare the selectively decellular xenoskin.The cytotoxicity was tested according to GB/T16886.5-2003 biological evaluation of medical devices for in vitro cytotoxicity,and the levels of cytotoxicity were evaluated with the United States Pharmacopeia.Subdermal implantation was tested according to GB/T16886.6-1997 biological evaluation of medical devices for local effects after implantation.Seventytwo mature Wistar rats were randomly assigned to groups A,B,and C（n=24）.Three kinds of materials were implanted into subcutaneous of rats back.Selectively decellular xenoskin was transplanted into group A,fresh porcine skin was transplanted into group B,and allogeneic skin was transplanted into group C.The samples were collected to make the observation of gross and histology after 1,2,4,8,12,and 16 weeks.Results The cytotoxicity was proved to be f irst grade by biocompatibility test.The gross and histological observation of subdermal implantation：after implantation,the most severe inflammatory reactions were seen in group B which dispersion was very slow.Inflammatory reactions in groups A and C alleviated gradually.In groups A and C,there was an increased collagen fiber density and angiogenesis at late stage;the transplanted skin was gradually degraded and absorbed.In group B,no obvious degradation and absorption were observed.Conclusion Selectively decellular xenoskin,prepared with glutaraldehyde solution,trypsin,and detergent solution,possesses characteristics of integral skin structure and excellent biocompatibility,so it can be used as a new type substitute to repair the burn wound.

关 键 词：选择性去细胞异种皮 生物相容性 猪 

分 类 号：R318[医药卫生—生物医学工程] R622[医药卫生—基础医学]