胶原膜包裹及环孢素A对鸡羽根角蛋白移植物组织相容性的改善作用  

Collagen-gel coating or use of cyclosporine A improves histocompatibility of in vivo grafted chicken calamus keratin

作　　者：仇欣霞[1] 赵冰雷[1] 陈英华[1] 郭家松[1] 肖应庆[1] 董为人[1] 

机构地区：[1]南方医科大学组织学与胚胎学教研室,广东省广州市510515

出　　处：《中国组织工程研究与临床康复》2008年第26期5160-5163,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省自然科学基金资助项目(05004719)"鸡羽根角蛋白在体组织工程化神经"~~

摘　　要：背景:前期实验提出将鸡羽根角蛋白管作为神经桥接材料这一设想,发现机体组织对鸡羽根角蛋白有一定的免疫排斥作用,埋植材料周围组织中有大量炎性细胞出现。目的:观察经胶原膜包裹或应用免疫抑制剂后,鸡羽根角蛋白材料在体埋植对周围组织的影响。设计、时间及地点:随机对照动物实验,于2006-10/2007-01在南方医科大学组织学与胚胎学实验室完成。材料:选择SD大鼠42只,按随机数字表法分成5组,空白对照组6只,其余各组各9只。方法:参照胶原蛋白提取方法,由牛跟腱提取胶原蛋白。将经特殊处理的鸡羽根角蛋白埋植入大鼠竖脊肌组织中,鸡羽根角蛋白材料+环孢素A组大鼠材料埋植后2周腹腔注射环孢素A5mg/(kg·d);鸡羽根角蛋白材料浸泡环孢素A组将材料浸泡于2.5g/L环孢素A后埋植;胶原膜包裹鸡羽根角蛋白材料组应用胶原膜包裹材料后埋植;单纯鸡羽根角蛋白材料组仅埋植材料;空白对照组仅作切口,不埋植材料。主要观察指标:分别于材料埋植后2,4,8周取材,光镜下观察埋植材料的降解情况及对周围组织的影响,应用免疫组织化学方法检测周围组织巨噬细胞的浸润情况。结果:①材料埋植后各组大鼠一般状况良好,创口未见肿胀及渗液。②材料埋植后2,4,8周,材料埋植周围组织可见大量巨噬细胞,尤其是多核巨细胞分布于材料边缘,吞噬材料;炎性细胞数量较少,以鸡羽根角蛋白材料+环孢素A组为著。③材料埋植后2,4,8周各组材料埋植周围组织均可见CD68阳性表达细胞,其中4周时巨噬细胞阳性数比其他两个时点多(P<0.05)。结论:短期应用免疫抑制剂可显著改善鸡羽根角蛋白的在体组织相容性;鸡羽根角蛋白在体内可降解,巨噬细胞尤其是多核巨细胞参与了该过程。BACKGROUND： Previous studies have assumed to use chicken calamus albumenduct as nerve bridging material, and found body tissue developed immunologic rejection to chicken calamus keratin （CCK） with inflammatory cells in tissues surrounding embedded materials. OBJECTIVE： To observe the effect of in vivo grafted CCK by collagen gel coating or by use of immunosuppressant （cyclosporine A） on tissues. DESIGN, TIME AND SETTING： Randomized controlled animal trial was performed at Department of Histology and Embryology, Southern Medical University between October 2006 and January 2007. MATERIALS： Forty-two SD rats were randomized into four experimental groups （n=9） and control group （n=6）. METHODS： Collagen gel was extracted from bovine tendon according to the approach of reference. After specific treatment, CCK was implanted into erector muscle of spine. In CCK ＋ cyclosporine A （CsA） group, the rats were received daily intraperitoneal injection of CsA （5 mg/kg） for two consecutive weeks after CCK implantation; in CCK soaked in CsA group, CCK was soaked in CsA （2.5 g/L） solution for 48 hours before grafting; in CCK coated with collagen gel group, CCK coated with collagen gel was grafted; and in CCK group, only CCK was implanted. In control group, the rats were not implanted with any material. MAIN OUTCOME MEASURES： CCK degradation and its effect on the surrounding tissues were observed at 2, 4 and 8 weeks after grafting. Immunohistochemistry was performed to identify macrophages in filtration in the host tissues. RESULTS： General condition of postoperative rats in each group was well. No swelling or effusion was observed on the wound. Histological observation showed at 2, 4 and 8 weeks after CCK implantation, numerous macrophages, especially MNGC were found on the margin of CCK grafts. Only a few inflammatory cells were seen in the host tissues, especially CCK ＋ CsA group. Immunohistochemistry observation showed at 2, 4 and 8 weeks after CCK implantation, CD68-positive cells w

关 键 词：鸡羽根角蛋白 环孢菌素 胶原 巨噬细胞 

分 类 号：R617[医药卫生—外科学]