中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (4): 546-552.doi: 10.12307/2022.090

• 材料生物相容性 material biocompatibility • 上一篇    下一篇

钛植入体表面钽功能涂层制备及性能表征

陈  硕1，肖东琴1，李兴平1，2，冉  斌1，3，匙  峰4，张成栋1，邓  丽1，黄南翔1，刘  康1，冯  刚1，段  可3   

  1. 1川北医学院第二临床医学院·南充市中心医院组织工程与干细胞研究所，四川省南充市 637000；2成飞医院骨科，四川省成都市   610031；3西南医科大学附属医院骨与关节外科，四川省泸州市   646000；4西华师范大学组织修复材料工程技术协同创新中心，四川省南充市   637000
  • 收稿日期:2020-11-11 修回日期:2020-11-14 接受日期:2020-12-21 出版日期:2022-02-08 发布日期:2021-11-03
  • 通讯作者: 肖东琴，博士，副研究员，川北医学院第二临床医学院·南充市中心医院组织工程与干细胞研究所，四川省南充市 637000 冯刚，博士，教授，川北医学院第二临床医学院·南充市中心医院组织工程与干细胞研究所，四川省南充市 637000 段可，博士，副教授，西南医科大学附属医院骨与关节外科，四川省泸州市 646000
  • 作者简介:陈硕，男，1990年生，汉族，川北医学院在读硕士，主要从事骨科临床和骨生物材料基础研究。
  • 基金资助:
    国家自然科学基金(82002289)，项目负责人：肖东琴；四川省科技厅应用基础项目(2018JY0100)，项目负责人：冯刚；南充市市校合作科研专项资金(19SXHZ0236)，项目负责人：张成栋；南充市市校合作科研专项资金(18SXHZ0375)，项目负责人：邓丽；南充市市校合作科研专项资金(19SXHZ0230)，项目负责人：黄南翔；西南医科大学附属医院博士科研启动基金(019038)，项目负责人：段可；川北医学院国家自然科学基金预研项目(CBY19-YZ09)，项目负责人：冯刚；川北医学院国家自然科学基金预研项目(CBY19-YZ18)，项目负责人：肖东琴

Preparation and characterization of tantalum functional coating on titanium implant

Chen Shuo1, Xiao Dongqin1, Li Xingping1, 2, Ran Bin1, 3, Shi Feng4, Zhang Chengdong1, Deng Li1, Huang Nanxiang1, Liu Kang1, Feng Gang1, Duan Ke3   

  1. 1Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital; 2Department of Orthopedics, Chengfei Hospital; 3Department of Orthopedics and Arthrology, Affiliated Hospital of Southwest Medical University; 4Collaboration Innovation Center for Tissue Repair Material Engineering Technology, China West Normal University
  • Received:2020-11-11 Revised:2020-11-14 Accepted:2020-12-21 Online:2022-02-08 Published:2021-11-03
  • Contact: Xiao Dongqin, PhD, Associate researcher, Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China Feng Gang, MD, Professor, Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China Duan Ke, PhD, Associate professor, Department of Orthopedics and Arthrology, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Chen Shuo, Master candidate, Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 82002289 (to XDQ); the Application Fundamental Project of Sichuan Provincial Department of Science and Technology, No. 2018JY0100 (to FG); the Nanchong City-School Cooperative Scientific Research Special Fund, No. 19SXHZ0236 (to ZCD); the Nanchong City-School Cooperative Scientific Research Special Fund, No. 18SXHZ0375 (to DL); the Nanchong City-School Cooperative Scientific Research Special Fund, No. 19SXHZ0230 (to HNX); the Doctoral Research Fund of the Affiliated Hospital of Southwest Medical University, No. 019038 (to DK); the Pre-research Project of National Natural Science Foundation of North Sichuan Medical College, No. CBY19-YZ09 (to FG); the Pre-research Project of National Natural Science Foundation of North Sichuan Medical College, No. CBY19-YZ18 (to XDQ)

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摘要：

文题释义：
钽功能涂层：利用表面改性技术在生物材料表面制备钽氧化物薄膜，使材料表面具有与钽相似的理化性质和生物学特性。
骨整合：是骨组织与植入体表面之间直接有序的结构和功能连接，它是骨组织和植入体接触后发生一系列分子和细胞效应的结果，为评估植入体治疗效果的重要指标。
背景：钛合金因其良好的生物相容性被广泛应用于临床骨科，但其作为生物惰性材料缺乏骨诱导活性，易导致植入假体松动，因此有必要对钛植入体表面进行改性来增强其成骨活性。 
目的：利用溶胶凝胶法在钛表面制备钽功能涂层，并对涂层的理化性质及成骨性能进行表征。
方法：利用溶胶凝胶法在医用钛片表面制备钽功能涂层，采用扫描电镜及能谱分析对涂层的表面形貌及元素组成进行表征，通过接触角测试评估钛片、钽片、钽涂层的表面亲水性。将兔骨髓间充质干细胞分别接种于钛片、钽片、钽涂层上，利用扫描电镜观察材料表面细胞黏附形态，荧光染色观察材料表面细胞黏附及存活，CCK-8法检测细胞增殖活性，碱性磷酸酯酶显色及茜素红S染色评估材料表面细胞的成骨分化能力。
结果与结论：①扫描电镜显示，涂层表面均匀分布着大小一致的纳米级颗粒，且表面涂层均一，未见裂纹产生；元素分析结果显示，钽涂层表面的元素主要为Ta、O、Ti元素；钽涂层表面的亲水性优于钛片、钽片；②接种12 h后的扫描电镜显示，细胞在钛片和钽片表面的黏附形态相似，呈长梭形，向周边伸出少量丝状伪足；细胞在钽涂层表面呈铺展态黏附生长，向远处伸出大量丝状伪足并与相邻细胞连接；③接种72 h后的荧光染色显示，细胞在钽涂层表面几乎均呈铺展态黏附生长，黏附细胞的数量多于钛片、钽片，并且钽涂层表面的活细胞数量多于钛片、钽片(P < 0.05)；④CCK-8法检测结果显示，钽涂层表面的细胞增殖速率快于钛片、钽片(P < 0.05)；⑤钽涂层表面细胞的碱性磷酸酶含量和钙结节形成数量均多于钛片、钽片；⑥结果表明，钽涂层修饰后的钛表面更利于骨髓间充质干细胞的黏附及成骨分化。

https://orcid.org/0000-0002-6572-9236 (陈硕) ；https://orcid.org/0000-0002-9587-3482(肖东琴)

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

关键词: 骨, 植入体, 钛合金, 钽涂层, 溶胶-凝胶法, 成骨性能, 细胞黏附

Abstract: BACKGROUND: Titanium (Ti) alloy is widely used in clinical orthopedics, because of its good biocompatibility. However, as a biological inert material, it is lack of osteoinductive activity, which is easy to cause implant loosening. Therefore, it is necessary to modify the surface of titanium implant for enhancing its osteogenic activity.
OBJECTIVE: To prepare Tantalum (Ta) functional coating on titanium surface by sol-gel method, and characterize the physicochemical properties and osteogenic properties of the coating.
METHODS: Ta functional coating was prepared on the surface of medical Ti sheet by sol-gel method (named as Ti-Ta). The surface morphology and element composition of the coatings were characterized by scanning electron microscope and energy spectrum analysis. The surface hydrophilicity of Ti sheet, Ta sheet, and Ti-Ta coatings was evaluated by contact angle test. Rabbit bone marrow mesenchymal stem cells were co-cultured with the Ti, Ta and Ti-Ta samples. Cell adhesion morphology on the material surface was observed by scanning electron microscope. Cell adhesion and survival were observed by fluorescence staining. Cell proliferation activity was detected by CCK-8 method. Alkaline phosphatase staining and alizarin red S staining were used to evaluate the osteogenic differentiation ability of cells on the material surface.
RESULTS AND CONCLUSION: (1) Scanning electron microscope images showed that nanoparticles were uniformly distributed on the surface of Ti-Ta sample and no cracks appeared. The elemental analysis showed that elements on the coating were mainly composed of Ta, O, Ti. Ti-Ta surface had better hydrophilicity than that of Ti and Ta. (2) Twelve hours after seeding, cells spread and adhered to the surface of Ti-Ta, protruding a large number of filamentous pseudopodia to the distance and connecting with adjacent cells. For cells on the surface of Ti and Ta, they showed long fusiform shapes, with a small amount of filamentous pseudopodia extending around. (3) The fluorescence staining results of living/dead cells showed that at 72 hours after seeding, a large number of living cells were observed on all surfaces. The number of adherent cells was more than that of Ti sheet and Ta sheet, and the number of living cells on the surface of Ti-Ta coating was more than that on Ti sheet and Ta sheet (P < 0.05). (4) CCK-8 results showed that Ti-Ta had the strongest ability to promote cell proliferation                (P < 0.05). (5) The content of alkaline phosphatase and the number of calcium nodules on the surface of Ti-Ta surfaces were much higher than those on the Ti and Ta surfaces. (6) In conclusion, the Ti surface modified by Ta coating was more favorable for the adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells.

Key words: bone, implant, titanium alloy, tantalum coating, sol-gel method, osteogenic properties, cell adhesion

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