钛板表面壳聚糖-金纳米粒子/整合素β6复合物涂层的制备与性能分析

Preparation and performance analysis of Ch-AuNPs/ITGB6 coating on pure titanium surface

目的在粗糙纯钛板表面制备壳聚糖-金纳米粒子/整合素β6复合物(Ch-AuNPs/ITGB6)涂层,研究ChAuNPs/ITGB6对细胞成骨分化和抗炎能力的影响。方法氧化还原法制备Ch-AuNPs,分别加入等质量ITGB6质粒(pLV-ITGB6)和空白质粒(p LV-cs2.0),在粗糙纯钛板表面分别形成Ch-AuNPs/ITGB6涂层和Ch-AuNPs/pLV涂层。应用动态光散射法(DLS)、电泳光散射法(ELS)检测Ch-AuNPs的湿态性质,通过透射电子显微镜(TEM)及扫描电子显微镜(SEM)观察涂层形态,使用能量色散X射线光谱(EDX)分析涂层性质。将具有Ch-AuNPs/ITGB6及Ch-AuNPs/pLV涂层的钛板分别与成骨细胞前体细胞(MC3T3-E1)共培养48 h,记为Ch-AuNPs/ITGB6组和Ch-AuNPs/pLV组。应用Western Blot检测两组骨桥蛋白(OPN)、骨钙素(OCN)、肿瘤坏死因子-α(TNF-α)及白介素-6(IL-6)的蛋白表达水平。结果 Ch-AuNPs湿态直径大多为50~120 nm、分布均匀(多扩散系数PDI=0.544),携带弱正电荷(zeta电位主要位于0~10 mV),可与ITGB6质粒形成Ch-AuNPs/ITGB6包被于钛板表面。与Ch-AuNPs/pLV组相比,Ch-AuNPs/ITGB6组可提高细胞OCN、OPN的蛋白表达水平,降低TNF-α、IL-6的蛋白表达水平。结论 Ch-AuNPs/ITGB6可成功包被于钛板表面,且能够促进MC3T3-E1中成骨分化蛋白的表达,降低炎症因子的表达水平,为种植体表面生物修饰提供新思路。

Objective To form a stable chitosan-gold colloidal nanoparticle/integrin β6 plasmid complex(Ch-AuNPs/ITGB6)on the surface of rough pure titanium plate to study the effect of Ch-AuNPs/ITGB6 on cell osteogenesis and antiinflammatory. Methods Ch-AuNPs was prepared based on REDOX method. Ch-AuNPs and pLV-cs2.0/pLV-ITGB6 were mixed with equal weight. Both Ch-AuNPs/pLV and Ch-AuNPs/ITGB6 coating were formed on the rough surface of titanium plate by charge adsorption. The properties of this coating surface were identified by DLS,ELS,TEM,SEM and Ch-AuNPs/pLV and Ch-AuNPs/ITGB6 coated PT up to48 hours. The expression levels of osteopontin(OPN),osteocalcin(OCN),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected with Western Blot.Results The wet diameter of Ch-AuNPs were mostly50-120 nm with even distribution(PDI = 0.544),which carried weak positive charges(zeta potential was mainly at0-10 m V). It could form Ch-AuNPs/ITGB6 together with ITGB6 to coat titanium plate. Ch-AuNPs/ITGB6 could increase the levels of OCN and OPN while decrease levels of TNF-α and IL-6. Conclusion The successful coating of Ch-AuNPs/ITGB6 on the surface of titanium plate can promote the expression of osteogenic differentiation protein in MC3 T3-E1 and reduce the expression level of inflammatory cytokines,which provides implications for novel surface biological modifications of implants.