微弧氧化钛表面壳聚糖/antimiR-138涂层构建及其生物学性能

Construction and biological properties of chitosan/antimiR-138 on microporous titanium oxide surface

目的:在微弧氧化钛表面构建壳聚糖(CS)/antimiR-138涂层并研究其生物学性能。方法:用CS包裹antimiR-138形成CS/antimiR-138载体颗粒,通过硅烷偶联的方法将其加载至经微弧氧化(MAO)处理的钛表面。再将钛试件分为MAO表面加载CS/antimiR-138组、加载CS/antimiR-control组、加载硅烷偶联处理组(MAO-SGC)和未经MAO处理组(MAO)(n=5),用接触角测量仪测量其表面水接触角;SEM观察其表面微形貌;CCK-8分析其生物相容性;RT-PCR检测其对MSCs内miR-138及成骨相关基因表达的影响。结果:各组钛表面的水接触角为MAO组<MAO-SGC组<MAO-CS/miRNA组;SEM观察表明MAO-SGC组表面及孔洞内部均有一层膜状物质,表面较平坦,局部微突起,加载CS/miRNA后其表面孔洞的内部及周围微结构较粗糙,可见大面积的颗粒状物质沉积于表面;CCK-8测定结果表明,4组材料生物相容性无统计学差异(P>0.05);RT-PCR结果表明,MAO-CS/antimiR-138组表面细胞内miR-138的表达量明显低于其他3组(P<0.05),各成骨相关基因的表达量明显高于其他3组(P<0.05)。结论:在微弧氧化钛表面构建CS/antimiR-138涂层,生物相容性优良,可促进MSCs的成骨分化。

AIM： To fabricate CS/antimiR - 138 modified microporous surface on Ti implants. METH-ODS ：The CS/antimiR - 138 complexes were fabricated, and then loaded onto the MAO Ti surface by silane glutaralde- hyde coupling. The MAO titanium surface was respectively loaded by CS/antimiR - 138, CS/antimiR - control,4 si-lane glutaraldehyde coupling agent （ MAO - SGC ） and the naked MAO （ MAO） （ n = 5 ） . Water contact angle was measured. SEM was used to observe the surface morphology. Bone marrow mesenchymal stem cells （ MSCs） were cul-tured on the specimens. The cell viability was examined by CCK - 8 assay. The expressions of miR - 138 and osteo-genesis -related genes of the cells were evaluated by real - time PCR. RESULTS： The water contact angles of MAO, MAO - SGC and MAO - CS/antimiR - 138 groups increased in turn significantly. The cell viability was not significant-ly different among the groups （ P 〉 0. 05 ）. MAO - CS/antimiR - 138 group exhibited lower expression of miR - 138 （P 〈0. 05） and higher expression of the osteogenesis - related genes than the other 3 groups（P 〈0. 05） . CONCLU-SION ： Covalent bonding of CS/antimiR - 138 onto MAO titanium surface can improve osteogenic differentiation of MSCs without cytotoxicity.