Abstract: 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 anti-inflammatory. 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 EDX techniques. MC3T3-E1 cells were transfected by Ch-AuNPs/pLV and Ch-AuNPs/ITGB6 coated PT up to 48 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 mostly 50 - 120 nm with even distribution（PDI = 0.544），which carried weak positive charges（zeta potential was mainly at 0 - 10 mV）. 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 MC3T3-E1 and reduce the expression level of inflammatory cytokines，which provides implications for novel surface biological modifications of implants.

Key words: Ti-implant surface modification；coating detection；chitosan-gold colloidal nanoparticle/integrin β6 plasmid complex, Ch-AuNPs/ITGB6

摘要： 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Ch-AuNPs/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（pLV-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。

关键词: 钛种植体表面改性, 涂层检测, 壳聚糖-金纳米粒子/整合素&beta, 6复合物