NaOH消蚀法制备骨细胞外基质材料的实验研究

Experimental Study of the Preparation of the Bone Extracellular Matrix Material by NaOH-Macertion Method

目的 : 研制一各新型的天然骨移植替代材料 ,为其临床应用提供实验依据。方法 : 采用NaOH消蚀法制作家兔骨细胞外基质支架 ,扫描电镜观察 ;对支架行生物相容性实验 ,将支架埋植于兔背部肌肉内 ,分别于术后 1周、2周、4周、6周取出行组织学观察 ,X射线能谱分析埋植前、后化学元素组成的改变。结果 : (1)SEM观察 ,以NaOH消蚀处理后 ,骨组织中的细胞成分被彻底清除掉 ,细胞外基质成分 -胶原纤维及骨盐则维持原有形态及三维立体网状结构。 (2 )光镜下观察 ,支架周围有成纤维细胞及毛细血管增生 ,轻度淋巴浸润 ;支架结构随埋植时间渐稀疏、紊乱。 (3)X射线能谱分析结果 ,埋植后钙原子含量低于埋植前水平 (p <0 .0 0 1)。结论 : NaOH消蚀骨支架作为骨移植替代材料 ,其三维立体网状结构为成骨细胞发挥功能提供了有利空间 ,有可能作为骨组织工程中种子细胞的载体 ;骨盐的存在使其具有一定的力学强度 ,适宜修复较大节段的骨缺损 ;组织相容性好 ,在体内可降解吸收。

Objective:To a new natural grafting material for repairing segmental bony defects.Method: The material was mgde by immersing the shaft of radius of rabbits in an aqueous solution of NaOH,which was observed with scanning electron microscope(SEM),and analyzed for its chemical elements.Then it was implanted into the dorsal muscles of rabbits for tissue reactions in vivo.It was individual observed by histological examination 1 week,2 weeks,4 weeks,6 weeks after implantation.To analyzed the changes of the chemical elements between before and after implantation by X-Ray energy spectrum.Results: The cellular elements of the bone treated with NaOH-macertion were removed effectively and consistently.The extracellular matrix consisted by collagen fibrils and minerals was preserved in the natural position which can not only constitute the framework but also provide a microenvironment suitable for cells activities.The mineral can make the frame more strength in mechanics.Histological examination revealed that there was mearly no tissue reaction during the implantation and the grafted frames were absorbed.Numbers of calcium of the frames reduced obviously after the implantation(p<0.001).Conclusion: The three dimensions framwork of the grafted material which is prepared by NaOH-maceration provides profited space for the osteoblasts and is likely to be used as the carrier of seed cells in bone histology engineers.The minerals in the bone can make the frame more strength in mechanics.The material has a fine histocompatibility and can be absorbed.It is a excellent material for the reparation of the bigger segmental bony defects.