煅烧骨的生物相容性及细胞相容性评价  

Biocompatibility and cytocompatibility of calcined bone

作　　者：刘斌钰[1] 马晓红 李宁毅[3] 樊功为[3] 金晓明[3] 袁荣涛[3] 

机构地区：[1]山西大同大学医学院口腔教研室,山西省大同市037008 [2]大同市第二人民医院,山西省大同市037008 [3]青岛大学医学院,山东省青岛市266012

出　　处：《中国组织工程研究与临床康复》2008年第41期8055-8058,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30471899);山西大同大学青年科学研究基金项目(同大科字200805)~~

摘　　要：背景:煅烧骨的主要成分是高纯度的羟基磷灰石,其钙磷比值接近于人骨,且制作方法简单,价格低廉,受到广泛关注。目的:观察煅烧骨的生物相容性及其与骨髓间充质干细胞的细胞相容性,为组织工程骨重建颌骨缺损奠定一定基础。设计、时间及地点:观察性实验,2008-04在青岛大学医学院中心实验室及口腔研究室完成。材料:成年牛长骨骨骺端在箱式电阻炉内煅烧成煅烧骨。方法:将煅烧骨与第3代已经诱导的骨髓间充质干细胞复合,倒置显微镜和扫描电镜观察细胞相容性;应用溶血试验、凝血试验、急性毒性试验及皮下植入试验观察煅烧骨的生物相容性。主要观察指标:煅烧骨的孔隙率和超微结构;煅烧骨的生物相容性及其与骨髓间充质干细胞的细胞相容性。结果:煅烧后的松质骨块呈白色,其骨小梁结构完整,骨小梁的间隙为相互连通的孔隙,孔径主要在190～750μm,孔隙率为(80.39±1.40)%。骨髓间充质干细胞接种到煅烧骨后,倒置显微镜显示接种即刻细胞即充满煅烧骨网孔,24h可见大量细胞黏附于支架上,7d后细胞分泌大量细胞外基质,细胞与基质分界不清;扫描电镜显示接种1d可见少量细胞黏附与煅烧骨上,细胞多个伪足向四周伸出,接种7d可见大量细胞与胶原纤维覆盖于支架上,基本包埋支架。生物相容性试验显示煅烧骨无溶血现象,不引起凝血功能改变,无毒,皮下植入后动物伤口无感染、化脓,植入材料无排出现象,取材时见材料周围肌肉颜色、质地正常。结论:煅烧骨经高温煅烧后,可以彻底消除其抗原性,不会产生免疫排斥反应,细胞、组织、血液相容性及生物安全性好,可作为颌骨组织工程支架。BACKGROUND： The main component of calcined bone is highly purified hydroxyapatite and the ratio of calcium to phosphorus is close to the human bone. Calcined bone is easy to prepare and low to cost, thus it attracts more and more attentions. OBJECTIVE： To observe biocompatibility of calcined bone and its cellular compatibility to bone marrow mesenchymal stem cells （BMSCs）, and to lay a foundation for repairing jaw defect by tissue engineering bony remodeling. DESIGN, TIME AND SETTING： An observational study was performed in the central laboratory and Department of Stomatology, Qingdao University Medical College in April 2008. MATERIALS： Long bone osteoepiphysis of adult bovine was calcinated to calcined bone using box resistor-stove. METHODS： The third generation BMSCs was compounded with calcined bone; The cellular compatibility of calcined bone to BMSCs was observed using inverted microscope and scanning electron microscope; The biocompatibility of calcined bone was observed through hemolysis test, thrombotest, acute toxicity test and hypodermic implantation test. MAIN OUTCOME MEASURES： Porosity and ultrastructure of calcined bone; Biocompatibility of calcined bone and its compatibility to BMSCs. RESULTS： Calcined cancellated bone block was white and bone trabecularism was intact. The ventage of bone trabecularism was interconnecting and presented a pore size of 190-750μm, a porosity of （80.39±1.40）%. It was clear under inverted microscope that meshes were full of cells soon as BMSCs were inoculated on calcined bone and a great quantity of cells adhered on bracket at 24 hours; The cells secreted massive extracellular matrix after 7 days and boundary was not clear between cells and matrix. Scanning electron microscope results showed a small quantity of cells adhered on calcined bone and they stretched out many pseudopodia when BMSCs were inoculated on calcined bone at one day. The bracket was covered with a great quantity of cells and collagen fibers and it was mainly embedded at 7 days.

关 键 词：煅烧骨 细胞相容性 生物相容性 骨髓间充质干细胞 

分 类 号：R318[医药卫生—生物医学工程]