摘要
目的既拥有良好生物相容性,又具备足够机械强度的三维多孔支架依然是骨组织工程领域研究的热点。文中评价三维联通纳米氧化锆多孔支架作为细胞载体,供骨髓间充质干细胞(bone marrow stromal cells,BMSCs)附着及生长情况。方法制备纳米氧化锆多孔支架,对其孔隙结构及机械性能进行表征,溶血实验评价其生物安全性。提取成年beagle犬骨髓,分离、培养犬BMSCs,取第3代细胞,以1×106个/支架密度接种于多孔支架作为实验组,扫描电镜观察细胞早期附着、MTT法检测细胞在支架上增殖、PNPP法检测细胞在支架上表达碱性磷酸酶(alkaline phosphate,ALP)情况。结果研究所得支架内部结构表现为三维联通,孔隙率测定值为(92.667±0.324)%,压缩强度值高达4.38MPa;溶血率为2.19%,符合生物材料安全性标准。扫描电镜观察可见犬BMSCs在氧化锆多孔支架表面及内部均能黏附、伸展、增殖,MTT检测结果显示,与细胞接种第1天比较(A值为0.261±0.034),第3、6、9、12天细胞在支架上增殖迅速(A值分别为0.604±0.059,1.215±0.019,1.385±0.254,1.506±0.050),差异有统计学意义(P<0.01)。ALP表达检测结果显示,与实验组第1天比较(A值为0.016±0.003),第7、14天细胞在支架上表达ALP水平(A值分别为0.032±0.002,0.062±0.007)显著升高,差异有统计学意义(P<0.01)。结论研究制备的三维联通纳米氧化锆多孔支架具有良好的生物相容性,可以作为细胞载体,供犬BMSCs附着生长。
Objective Nowadays,in the field of bone tissue engineering,searching for a kind of scaffold,which is both strong enough and biocompatible,is still a hot topic.Objective of this study is to evaluate the three-dimensional nano-zirconia porous scaffold’s effect on bone marrow stromal cells early attachment and proliferation.Methods Preparing nano-zirconia porous scaffold and testing its porosity and its mechanical strength.Hemolysis experiment was taken to evaluate its biosafety.Canine bone marrow stromal cells（ BMSCs） were extracted from an adult beagle and cultured in vitro.The third generation cells were seeded onto the scaffold and induced to osteoblasts under condition medium.Cells early attachment on scaffold was observed by SEM.MTT and ALP tests were taken to evaluate cells proliferation and osteogenesis within two weeks.Results Nano-zirconia porous scaffold in our study presented a porous structure with a well connected inner.Its porosity and compressive strength were（ 92.667 ±0.324） % and 4.38MPa,respectively.Hemolysis rate was2.19%,which was demonstrated to be safe to human body.Canine BMSCs could colonize the surface of the materials,bridge macro-porosity and proliferate on the scaffold.A values taken by the method of MTT showed that cells could proliferate quickly on the scaffold after 3,6,9,12 d（ A values were 0.604 ± 0.059,1.215 ± 0.019,1.385 ±0.254,and 1.506 ± 0.050）,comparing with that on the first day（ A value was 0.261 ± 0.034）.Statistical difference was found（ P ＆lt;0.01）.The ALP level raised up significantly on day 7 and 14（ A values were 0.032 ± 0.002 and 0.062 ± 0.007）,comparing with that on the first day（ A value was 0.016 ± 0.003）.Statistical difference was found（ P ＆lt; 0.01）.Conclusion The three-dimensional nano-ZrO2porous scaffold in our study presented satisfactory biocompatibility.It could offer a microenvironment for osteoblasts growth and secreted osteogenesis marker in vitro.
出处
《医学研究生学报》
CAS
北大核心
2014年第6期564-567,共4页
Journal of Medical Postgraduates
基金
国家自然科学基金(81071261
30970722)
全军科技创新项目(cws11c016)
南京军区重点课题资助(10Z028)
关键词
骨组织工程
氧化锆
骨髓间充质干细胞
Bone tissue engineering
Nano-ZrO2
Bone marrow stomal cell
