钛表面载铜抗菌功能膜的制备及性能  

Preparation and properties of copper-loaded antibacterial functional film on titanium surface

作　　者：李兴平 肖东琴[2] 赵桥 陈硕 白亦光[2] 刘康[2] 冯刚 段可[1] Li Xingping;Xiao Dongqin;Zhao Qiao;Chen Shuo;Bai Yiguang;Liu Kang;Feng Gang;Duan Ke(Department of Orthopedics,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Institute of Tissue Engineering and Stem Cell,Nanchong Central Hospital,the Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Orthopedics,Chengfei Hospital,Chengdu 610031,Sichuan Province,China)

机构地区：[1]西南医科大学附属医院骨科,四川省泸州市646000 [2]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 [3]成飞医院骨科,四川省成都市610031

出　　处：《中国组织工程研究》2021年第4期553-557,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81171472),项目负责人:冯刚;四川省科技厅应用基础项目(2016JY0123),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;四川省卫计委科研项目(18PJ476),项目参与者:肖东琴;南充市市校合作科研专项资金(NSMC20170310),项目负责人:冯刚;南充市市校合作科研专项资金(19SXHZ0099,19SXHZ0236),项目参与者:肖东琴;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。

摘　　要：背景:钛金属植入物凭借其较好的组织相容性及生物惰性被广泛运用于当代骨科领域,但单纯的钛植入物难以满足复杂的临床需求,如细菌感染、骨再生速度慢等,故亟需开发具有生物功能的钛植入物。目的:鉴于铜离子在抗菌方面发挥的重要作用,在钛金属表面制备载铜功能膜,评价其体外抗菌功能及细胞活性。方法:通过浸涂方式在钛片表面制备含不同质量浓度铜离子(0,0.1,0.5,1.0 g/L,依次设定为B、C、D、E组)的聚乳酸薄膜。将金黄色葡萄球菌、大肠杆菌分别与纯钛片(A组)及4种载聚乳酸膜钛片共培养,大体与扫描电镜观察细菌菌落形成及黏附情况,活/死细菌染色观察细菌状态。将骨髓间充质干细胞分别与5组钛片(或5种浸泡DMEM培养基6 h后的钛片样品)共培养,采用CCK-8法评价细胞增殖能力。结果与结论:①大体观察显示,A、B组培养皿内的菌落数量最多,且均匀分布;C、D组靠近钛片区域的菌落数量减少,远离钛片区域的菌落数量较多,E组未见明显菌落形成;扫描电镜显示,A、B组表面黏附大量的细菌,C、D、E组表面黏附的细菌逐渐减少;②活/死细菌染色显示,A、B组细菌存活率均在95%以上,C、D、E组细菌存活率均在5%以下,且随着膜内铜离子浓度的增加细菌存活率降低;③细胞与钛片接触共培养的CCK-8检测显示,与A组比较,C组骨髓间充质干细胞增殖明显,D、E组细胞增殖受到抑制;细胞与浸泡后钛片样品共培养的CCK-8检测显示,与A组比较,C、D、E组细胞增殖明显;④结果表明,当聚乳酸溶液中铜离子质量浓度为0.1 g/L时,在钛金属表面制备的聚乳酸薄膜促细胞增殖能力最强,但杀菌能力相对较弱;在铜离子质量浓度为1.0 g/L时,聚乳酸薄膜的杀菌能力最强,DMEM浸泡6 h后无细胞毒性。因此,在钛片表面制备含适宜浓度的载铜聚乳酸薄膜能赋予植入体表面抗菌性能。BACKGROUND:Titanium implants are widely used in the field of contemporary orthopedics due to their good histocompatibility and biological inertia.However,titanium implants alone are difficult to meet complex clinical needs,such as bacterial infections and slow bone regeneration,so it is urgent to develop titanium implants with biological functions.OBJECTIVE:To evaluate its antibacterial function and cellular activity in vitro,the cupride-carrying functional membrane was prepared on the surface of titanium in view of the important role played by copper ions in antibacterial activities.METHODS:Poly(lactic acid)films containing different concentrations of copper ions(0,0.1,0.5,1.0 g/L,and assigned to groups B,C,D,and E)were prepared on the surface of the titanium sheet by dipping.Staphylococcus aureus and escherichia coli were co-cultured with pure titanium tablets(group A)and four kinds of poly(lactic acid)coated titanium tablets.The formation and adhesion of bacterial colonies were observed by scanning electron microscopy.Living/dead bacteria were stained to observe the bacterial state.Bone marrow mesenchymal stem cells were cultured with five groups of titanium tablets(or five kinds of titanium samples after immersion in DMEM for 6 hours).Cell proliferation was evaluated by CCK-8 method.RESULTS AND CONCLUSION:(1)General observation showed that the number of bacterial colonies in the petri dish was largest and they were evenly distributed in groups A and B.The number of bacterial colonies close to the titanium sheet area decreased,while the number of bacterial colonies far away from the titanium sheet area was large in groups C and D.No obvious colony formation was found in group E.Scanning electron microscopy showed that a large number of bacteria adhered to the surface of groups A and B,while the number of bacteria adhered to the surface of groups C,D and E gradually decreased.(2)Living/dead bacteria staining showed that the bacterial survival rate in groups A and B was above 95%,and that in groups C,D and E was

关 键 词：骨 材料 钛 骨髓炎 抗菌 铜离子 植入物 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]