脱矿骨在免疫抑制大鼠体内的骨诱导活性  

Osteoinductivity of demineralized bone implants in immuno-suppressed rats

作　　者：陈学英[1,2] 李宝兴[1,2] 李靖[3] 殷少芳[3] 

机构地区：[1]南方医科大学解剖学教研室 [2]山西省医用组织库,山西省太原市030006 [3]山西省医用组织库

出　　处：《中国组织工程研究与临床康复》2008年第23期4410-4413,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国防民基项目(Y05-020403-QS)~~

摘　　要：背景:传统的脱矿骨骨诱导活性检测方法是在小鼠股部肌袋异位植入模型,通过异位成骨方式评价骨诱导物质活性的高低,但由于动物宿主对植入材料存在着免疫排斥,影响了骨诱导活性的表达。目的:实验旨在建立以免疫抑制大鼠为实验动物骨诱导活性检测的新方法,降低动物对植入材料的免疫排斥,从而不影响其骨诱导活性的表达。设计、时间及地点:独立样本观察实验,于2005-09/2006-04在山西省医用组织库完成。材料:合格供体长骨皮质骨用于脱矿骨的制备,体质量200g左右的Wistar大鼠24只。方法:采用超声条件下盐酸脱矿、冻干、辐照等方法,制备人类脱矿皮质骨,对照样品为经高温高压灭活的脱矿骨。Wistar大鼠腹腔注射地塞米松造成免疫抑制,手术形成双侧股部肌袋,分别植入试验样品与对照样品。主要观察指标:术后3、4、5、6周取出植入材料行组织学观察与碱性磷酸酶的活性检测。结果:组织学观察显示,术后3周实验组脱矿骨吸收腔内间充质细胞聚集,术后4周脱矿骨内开始形成软骨细胞与软骨基质,术后5周形成较多的软骨组织,术后6周时形成类骨质。对照组材料被纤维组织包裹,显示吸收,未见成骨迹象。实验组碱性磷酸酶活性显著高于对照组(P<0.05)。结论:采用腹腔注射地塞米松方法造成大鼠免疫抑制,降低了宿主大鼠对植入人脱矿骨基质的排斥,异位植入后,脱矿骨吸收腔隙内出现软骨细胞,形成软骨组织与类骨质。BACKGROUND： Traditionally the osteoinductivity of demineralized bone is tested through ectopic implantation into thigh muscle pouches of mice. The expression of osteoinductivity is influenced due to immunologic rejection of the receptor animals. OBJECTIVE： To study a new method to evaluate the osteoinductivity of demineralized bone in the immuno-suppressed rats, reduce the immunologic rejection of the receptor animals, and ensure the expression of osteoinductivity. DESIGN, TIME AND SETTING： Independent sample observation was performed at the Shanxi Provincial Tissue Bank （Taiyuan Shanxi, China） from September 2005 to April 2006. MATERIALS： Demineralized bone was obtained from cortical bone of qualified donor. Twenty-four Wistar rats aged 200 g were used as experimental animals. METHODS： Demineralized bone was prepared from long bone of human by a series of treatments including demineralization with hydrochloric acid, freeze-drying and irradiation. Samples of control group were made from demineralized bone by autoclave treatment. Dexamethasone was intraperitoneally injected into Wistar rats in order to suppress their imunno-response. Intramuscular bone transplantation was performed by implanting the demineralized bone into bluntly dissected muscle groups of the rats. MAIN OUTCOME MEASURES： Histological observation was performed at 3, 4, 5, 6 weeks post-operatively. Alkaline phosphatase activity of the implanted samples was also tested. RESULTS： Histological observation showed that, mesenchymal cells migrated into the absorption lacuna of the implants of demineralized bone group at 3 weeks post-operatively, chondrocytes and cartilage matrix could be seen within the demineralized bone implants at 4 weeks, more cartilage tissues formed at 5 weeks, and osteoid was presented at 6 weeks. No bone formation was detected in the implants of control group post-operatively. Alkaline phosphatase activity of the demineralized bone group was significantly higher than that of the control group （P �

关 键 词：脱矿骨 骨诱导活性 免疫抑制 

分 类 号：R318[医药卫生—生物医学工程]