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目的 构建一种兼具"纳米形貌调控+表面化学功能化"的超亲水钛涂层，评价其理化性能及其对骨髓间充质干细胞(BMSCs)黏附铺展、增殖和成骨分化的影响。方法 恒电压条件下对钛表面进行阳极氧化制备TiO2纳米管结构，随后在纳米管表面引入丙烯酸以获得丰富–COOH位点，并通过偶联反应进一步接枝2，2′-氧双(乙胺)，构建含–CONH–及末端–NH2的超亲水钛功能涂层。根据样品制备过程，将样品分别分为未处理组、阳极氧化组、阳极氧化后接枝丙烯酸组和阳极氧化后接枝丙烯酸再接枝2，2′-氧双(乙胺)组，以上各组分别用Ti、TNT、TNT-COOH、TNT-NH2表示。采用场发射扫描电镜观察样品形貌特征；通过接触角评价表面润湿性；通过表面ζ电位分析仪测定生理条件下(pH7.4)表面ζ电位来表征表面电性；结合傅里叶变换红外光谱仪(FTIR)与X射线光电子能谱仪(XPS)分析表面元素组成和化学键变化。采用细胞计数试剂盒(CCK-8)法与细胞染色评价生物相容性，在成骨诱导条件下培养，分别通过碱性磷酸酶染色及活性定量、茜素红S染色评价矿化结节形成，从而评估不同表面改性对成骨分化的作用。结果成功构建了“阳极氧化TiO2纳米管+丙烯酸引入–COOH + 2，2′-氧双(乙胺)接枝”的功能化超亲水钛种植体表面涂层。FTIR与XPS结果证实表面–COOH位点成功引入并进一步形成酰胺键结构；改性后样品表面润湿性显著改善，呈超亲水特征，且表面ζ电位发生相应改变。体外实验CCK-8与细胞染色结果显示，与对照组相比，改性组细胞黏附与铺展更充分，细胞活力/增殖水平更佳且未见明显细胞毒性；在成骨诱导条件下，改性组碱性磷酸酶(ALP)表达/活性增强，矿化结节形成增加，表明该表面改性可有效促进早期成骨分化。结论基于阳极氧化在钛表面构建TiO2纳米管结构，并通过丙烯酸引入–COOH位点，进一步接枝2，2′-氧双(乙胺)获得末端–NH2的功能化超亲水表面，成功实现了钛表面“形貌+化学”的协同调控。该改性表面在体外表现出良好的细胞相容性，可促进BMSCs细胞黏附铺展与增殖，并增强成骨相关分化与矿化能力。
Abstract：Objective To construct a superhydrophilic titanium coating that integrates nanotopographical modulation with surface chemical functionalization， and to evaluate its physicochemical properties as well as its effects on the adhesion and spreading， proliferation， and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods Titanium surfaces were anodized under a constant-voltage condition to fabricate TiO2 nanotubular structures. Acrylic acid was subsequently introduced onto the nanotube surface to provide abundant –COOH groups， followed by further grafting of 2，2′-oxybis(ethylamine) through a coupling reaction， thereby constructing a superhydrophilic functional titanium coating containing –CONH– linkages and terminal –NH2 groups. According to the sample preparation procedure， the specimens were divided into four groups: untreated titanium (Control)， anodized titanium (AO)， anodized titanium grafted with acrylic acid (AA)， and anodized titanium grafted with acrylic acid followed by 2，2′-oxybis(ethylamine) coupling (AEEA). Surface morphology was characterized by field-emission scanning electron microscopy. Surface wettability was evaluated by contact angle measurement， and the surface zeta potential under physiological conditions (pH = 7.4) was determined using a surface zeta potential analyzer to characterize surface charge properties. Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) were further employed to analyze surface elemental composition and chemical bonding changes. Biocompatibility was assessed by the Cell Counting Kit-8 (CCK-8) assay and cell staining. Under osteogenic induction conditions， alkaline phosphatase staining and activity quantification， as well as Alizarin Red S staining， were performed to evaluate mineralized nodule formation， thereby assessing the effects of different surface modifications on osteogenic differentiation. Results The present study successfully constructed a functionalized superhydrophilic titanium implant surface coating based on anodized TiO2 nanotubes， followed by acrylic acid-mediated introduction of –COOH groups and subsequent grafting of 2，2′-oxybis(ethylamine). FTIR and XPS analyses confirmed the successful introduction of surface –COOH sites and the subsequent formation of amide bond structures. After modification， the surface wettability of the samples was markedly improved， exhibiting a superhydrophilic characteristic， accompanied by corresponding changes in surface zeta potential. In vitro CCK-8 and cell staining results showed that， compared with the control group， the modified surfaces promoted more sufficient cell adhesion and spreading， enhanced cell viability/proliferation， and exhibited no obvious cytotoxicity. Under osteogenic induction conditions， the modified groups demonstrated increased ALP expression/activity and enhanced mineralized nodule formation， indicating that this surface modification could effectively promote early osteogenic differentiation. Conclusions TiO2 nanotube arrays were constructed on titanium surfaces by anodic oxidation， –COOH sites were introduced via acrylic acid grafting， and 2，2′-oxybis(ethylamine) was subsequently coupled to obtain a functionalized superhydrophilic surface terminating in –NH2 groups， thereby achieving synergistic regulation of both topography and surface chemistry on titanium. In vitro， the modified surface exhibited favorable cytocompatibility， promoting the adhesion， spreading， and proliferation of BMSCs and enhancing osteogenesis-related differentiation and mineralization.
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基本信息:
中图分类号:R318.08
引用信息:
[1]陈雅庆,曹颖,郑顺丽.超亲水酰胺/氨基功能化钛涂层促成骨分化研究[J].安徽医科大学学报().
基金信息:
国家自然科学基金项目（编号：82101070）; 安徽医科大学博士科研资助基金项目（编号：XJ201918）;安徽医科大学口腔医学院学科建设项目（编号：2022xkfyts09）
2026-08-26
2026-08-26
2026-08-26