Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (4): 553-557.doi: 10.3969/j.issn.2095-4344.2368

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

Li Xingping1, 2, 3, Xiao Dongqin2, Zhao Qiao1, 2, Chen Shuo2, Bai Yiguang2, Liu Kang2, Feng Gang1, 2, Duan Ke1   

  1. 1Department of Orthopedics, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Institute of Tissue Engineering and Stem Cell, Nanchong Central Hospital, the Second Clinical Medical College of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; 3Department of Orthopedics, Chengfei Hospital, Chengdu 610031, Sichuan Province, China
  • Received:2020-02-18 Revised:2020-02-26 Accepted:2020-03-30 Online:2021-02-08 Published:2020-11-21
  • Contact: Feng Gang, MD, Professor, 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 Duan Ke, MD, Associate professor, Department of Orthopedics, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Li Xingping, Master candidate, 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
  • Supported by:
    the National Natural Science Foundation of China, No. 81171472; the Applied Foundation Project of Science and Technology Department of Sichuan Province, No. 2016JY0123, No. 2018JY0100; the Scientific Research Project of Sichuan Provincial Health and Family Planning Commission, No. 18PJ476; the Nanchong City School Cooperation Research Special Fund, No. NSMC20170310, No. 19SXHZ0099, No. 19SXHZ0236; the Foundation for Doctoral Research of Affiliated Hospital of Southwest Medical University, No. 019038

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Abstract: 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 below 5%. The bacterial survival rate decreased with the increase of copper ion concentration in the membrane. (3) CCK-8 detection of cells in contact culture with titanium tablets showed that compared with group A, the proliferation of bone marrow mesenchymal stem cells in group C was obvious, and the proliferation of cells in groups D and E was inhibited. CCK-8 detection of cells cultured with titanium slices showed that compared with group A, the proliferation of cells in groups C, D and E was significant. (4) The results showed that when the copper ion content in poly(lactic acid) solution was 0.1 g/L, the poly(lactic acid) film prepared on the surface of titanium had the strongest cell proliferation promoting capacity, but the bactericidal capacity was relatively weak. When the ion content was 1.0 g/L, the bactericidal capacity was the strongest. DMEM immersion for 6 hours showed no cytotoxicity. Therefore, the preparation of copper-loaded poly(lactic acid) film with appropriate concentration on the surface of titanium sheet can endow the antibacterial properties of the implant surface.

Key words: bone, materials, titanium, osteomyelitis, antibacterial, copper ions, implants

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