生物材料Sapeptide的体外塑型及在体毒性实验  

Fabrication in vitro and cytotoxic test in vivo of self-assembling peptides

作　　者：赵玺龙[1] 赵熠[2] 徐文莽[1] 李涛[1] 蔡琳[1] 

机构地区：[1]解放军成都军区昆明总医院病理实验科,云南省昆明市650032 [2]解放军成都军区昆明总医院电镜中心,云南省昆明市650032

出　　处：《中国临床康复》2006年第25期52-54,i0003,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(30171068)~~

摘　　要：目的:观察生物材料Sapeptide的大体形状及其表面结构特性,并进行在体毒性实验,评测其生物相容性。方法:实验于2004-08/2005-09在解放军第三军医大学外科研究所完成。①采用等渗盐水作为Sapeptide塑型剂,塑型后分别进行大体、光镜和电镜观察材料表面结构。②对成年Wistar大鼠分组进行腿部肌肉注射毒性实验。取成年Wistar大鼠4只,标记后分为2组。A组:左前肢为RAD16-Ⅱ,右前肢为RAD16-Ⅰ,左后肢为RAD16-Ⅱ和RAD16-Ⅰ的均等合剂,右后肢为生理盐水对照。B组:左前肢为RAD16-Ⅱ,左后肢为RAD16-Ⅰ,右前肢和右后肢为1.25g/L胰酶作对照。其中RAD16-Ⅱ和RAD16-Ⅰ水溶液浓度为2g/L,每个肢体注射液体量均为20μL。每组分为两个时相点(9d和35d)用剪刀按照标记部分取出被注射的整条肌肉组织,切片后做苏木精-伊红染色和刚果红染色,观察肌肉组织结构的改变及有无炎症反应。结果:①Sapeptide的大体形状与表面结构:Sapeptide材料表面呈网状,网孔直径约54μm,纤维的直径约为9μm。②毒性实验结果:大体观察,各组标本材料植入部位未见出血、积液和组织坏死,注射部位肌肉组织质地光泽正常;刚果红染色,可见所有注射Sapeptide材料的肢体切片中均有片状的红色物质,9d时面积大于35d所见;生理盐水对照组和胰蛋白酶组未见红染物质;苏木精-伊红染色,A组,第9天和第35天四肢肌肉组织取材标本见肌肉纤维形态正常,未见炎性细胞浸润;B组,注射胰蛋白酶侧肢体在第9天可见局部组织有炎症反应,见少量淋巴细胞浸润,但第35天,组织炎症已基本消退。Sapeptide注射处肌肉组织未观察到炎症反应和免疫排斥反应。结论:Sapeptide材料具有生物相容性好,可同种子细胞和宿主组织很好的整合;合适的生物可降解性,降解产物对人体无不良反应,有良好的可塑性;并且应用简便,在神经组织工程中是一种具有良好特性的组�AIM： To observe the gross shape and surface structure characters of selfassembling peptides （Sapeptides）, and evaluate its biocompatibility following cytotoxic test in vivo. METHODS： This experiment was carried out at the Research Institute of Surgery, Third Military Medical University of Chinese PEA from August 2004 to September 2005. （1）The Sapeptides solution were exposed in the isotonic saline solution. After they had been fabricated, the surface texture of material was observed separately under optical microscope and electron microscope. （2）The cytotoxicity test of Sapeptide in vivo was carried out through muscle injection in rats. Four adult Wistar rats were randomly divided into two groups with 2 rats in each group. Group A： Injected RAD16-Ⅱ Sapeptide into the left foreleg, RAD16-Ⅰ Sapeptide in the right foreleg, two Sapeptides mixture solutions in the left hind leg, and the same volume of physiological saline in the right hind leg was used for parallel control. Group B： RAD16-Ⅱ Sapeptide in the left foreleg, BAD16- Ⅰ sapeptide in the left hind leg, and 1.25 g/L of trypsin in the right hind leg was set as control. Among them, the concentration of water solution of RAD16-Ⅱ andRAD16- Ⅱ both was 2 g/L, 20 μL solution was injected into each limb. Each group was further divided into 2 subgroups according to different time points （the 9^th and 35^th day after injection）. The entire muscle tissue, which was labeled, was dissected and sectioned. The muscle sections were then stained with hematoxylin-eosin. Change of muscular tissue structure and whether inflammatory reaction existed were observed. RESULTS：（1） Gross shape and surface structure of Sapeptide： The surface of Sapeptide scaffolds was reticulate, the diameter of its hiatus was 54 p.m, and Sapeptide fibers were approximately 9 μm in diameter. （2）Cytotoxic test： With naked-eye observation, neither necrosis nor hemorrhage could be seen around Sapeptides. With Congo red, the Sapeptides were stained to red f

关 键 词：组织工程 支架 生物相容性材料 毒性试验 

分 类 号：R318.08[医药卫生—生物医学工程]