武汉云克隆科技股份有限公司

Abstract: To mimic the extracellular microenvironment of bone, a bioactive multilayered structure of gelatin/chitosan pair, containing bone morphogenetic protein 2(BMP2) and fibronectin (FN), was constructed onto Ti6Al4V surface via a layer-by-layer assembly technique. The successful fabrication of multilayered structure was confirmed by contact angle measurement, field emission scanning electron microscopy (FE-SEM) and confocal laser scanning microscopy (CLSM), respectively. Bioactive BMP2 released in a sustained manner along with the degradation of multilayered structure. MSCs grown onto the multilayer coated TC4 substrates displayed significantly higher (p<0.01 or p<0.05) production levels of alkaline phosphatase (ALP), mineralization and genes expressions of runt related transcription factor 2 (Runx2), osterix, osteocalcin (OC), osteopontin (OPN), ALP and collagen type I(Col I) compared to the controls after culture for 7 days and 21 days, respectively. More importantly, Micro CT analysis and histological observations demonstrated that the multilayer coated Ti6Al4V implants in vivo promoted the bone density and new bone formation around them after implantation for 4 weeks and 12 weeks, respectively. The results indicated that Ti6Al4V coated with biofunctional multilayers was beneficial for osteogenesis and integration of implant/bone. The study therefore presents an alternative to fabricate bio-functionalized Ti6Al4V-based implants for potential application in orthopedics field.

摘要：为了模拟骨的细胞外微环境，通过逐层组装技术将包含骨形态发生蛋白2（BMP2）和纤连蛋白（FN）的明胶/壳聚糖对的生物活性多层结构构建到Ti6Al4V表面上。通过接触角测量，场发射扫描电子显微镜（FE-SEM）和共聚焦激光扫描显微镜（CLSM）分别证实了多层结构的成功制备。随着多层结构的降解，生物活性BMP2以持续方式释放。在多层包被的TC4底物上生长的MSC表现出显着高于碱性磷酸酶（ALP）生成水平（p <0.01或p <0.05），矿化和基因表达的runt相关转录因子2（Runx2），osterix，骨钙素（OC）骨桥蛋白（OPN），ALP和I型胶原（Col I）分别与培养7天和21天后的对照进行比较。更重要的是，显微CT分析和组织学观察表明，植入多层包衣的Ti6Al4V体内植入物分别在植入4周和12周后促进了骨密度和围绕它们的新骨形成。结果表明，涂覆有生物功能多层膜的Ti6Al4V有利于植入物/骨的成骨和整合。因此，该研究提出了一种替代方案来制造可用于骨科领域的生物功能化的基于Ti6Al4V的植入物。

使用试剂原文信息：Osteocalcin assay kit and BMP2 quantikine ELISA Kit were provided by Uscnlife Science and Technology Co., Ltd. (Wuhan, China) and R&D System, Inc. (USA), respectively.