去除移植免疫反应抗原成分脱细胞天然骨基质的制备  

Preparation of acellular natural bone matrix to remove immunological rejection antigen

作　　者：李康杰[1] 孙抒[2] 

机构地区：[1]延边大学附属医院骨二科,吉林省延吉市133000 [2]延边大学医学部病理学教研室,吉林省延吉市133000

出　　处：《中国组织工程研究与临床康复》2011年第8期1355-1359,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81000791);仿生化衍生骨修复骨缺损的研究~~

摘　　要：背景:同种异体骨来源广泛,便于大量加工贮存,如能去除其免疫排斥反应用于骨支架修复骨缺损,将很好地解决骨源问题,但目前去除免疫排斥的方法不尽理想。目的:运用低渗-脱细胞联合过氧化氢去除骨基质中引起免疫排斥反应的抗原成分。方法:将SD大鼠股骨经低渗-脱细胞、过氧化氢处理制备天然脱细胞基质,再通过苏木精-伊红和甲苯胺蓝染色、扫描电镜与透射电镜观察、有机成分分析、洗脱剂残留量测定和移植免疫分析方法评估处理效果,并与新鲜大鼠股骨标本作比较。结果与结论:组织学观察可见处理后的脱细胞骨基质中胶原纤维排列规则,骨陷窝空虚,未见细胞,纤维连接蛋白和层粘连蛋白多存留在骨陷窝周边部。扫描电镜观察见处理后的骨基质板层连接疏松,板层之间出现大量孔隙;透射电镜见骨陷窝区已无高密度影;高效液相色谱仪检测TritonX-100在脱细胞骨基质中几无残留。光镜及扫描电镜均可见新鲜骨中有大量细胞成分。淋巴细胞刺激实验表明新鲜骨引起的免疫排斥反应明显强于脱细胞骨基质(P < 0.01)。说明联合应用低渗-脱细胞和过氧化氢处理基质的方法去除免疫排斥反应较为理想。BACKGROUND：Allograft implants have a variety of sources and are easy to be processed and stored.If the immunological rejection is able to be removed,they will be applied as bone scaffolds to repair bone defect,which will resolve human bone supply ultimately.But there is no perfect approach to avoid immunological rejection at present.OBJECTIVE：To remove antigen leading to immunological rejection in acellular natural bone matrix with hypotension-decellularization combined with hydrogen peroxide.METHODS：SD rat femurs were processed by hydrogen peroxide handling following hypotonic-decellularized procedures.Then,the manufactured acellular natural bone matrix were evaluated by hematoxylin-eosin staining,toluidine blue staining,scanning electron microscope,transmission electron microscope,organic component analysis,eluant residue measurement and transplantation immunity analysis.The femoral specimens were compared with fresh rat femur.RESULTS AND CONCLUSION：Histological observation showed that collagen fibers of acellular natural bone matrix arranged regularly and bone lacuna were lack of osteocytes,the fibronectin and laminin were present mainly in space around lacuna.Scanning electron microscope displayed that different layers of acellular natural bone matrix connected loosely and presented amounts of pore space between layers.Transmission electron microscope showed that no high density image appeared in lacuna area.There was no TritonX-100 in acellular natural bone matrix with the measurement of high-performance liquid chromatography.Light microscopy and scanning electron microscopy observations showed a large amount of cellular components were seen in fresh bone.Lymphocyte stimulation assay showed that immunological rejection of flesh bone was much greater than that of acellular natural bone matrix（P 0.01）.Above results indicated that antigen leading to immunological rejection could be removed mostly with procedures of hypotonic-decellularized processing and hydrogen peroxide handling.

关 键 词：天然脱细胞骨基质 低渗-脱细胞 过氧化氢 免疫排斥反应 生物材料 

分 类 号：R318[医药卫生—生物医学工程]