In vitro antibacterial activity and biological safety of nano-silver composite coating on the surface of titanium base

doi:10.3969/j.issn.2095-4344.2014.52.005

Abstract

Abstract:

BACKGROUND: Hydroxyapatite is the main component of bone with the capacity of osteogenic induction. Nano-silver has broad-spectrum and efficient antibacterial activity, while hydroxyapatite/nano-silver composite coating on the surface of titanium base has either biological activity or antimicrobial properties.

OBJECTIVE: To study the antimicrobial properties of hydroxyapatite/nano-silver composite coating on the surface of titanium base and to evaluate the biocompatibility and security of hydroxyapatite/nano-silver composite coating on the surface of titanium base.

METHODS: Silver solution of 0, 0.5, 1.0 mmol/L was selected to prepare hydroxyapatite/nano-silver composite coating samples deposited on the surface of titanium base. Then, Staphylococcus aureus was cultured in the sample leaching solution for the in vitro antibacterial qualitative analysis. In accordance with national and international standardization for biocompatibility testing, healthy adult Kunming mice and New Zealand rabbits were subjected to pyrogen test, hemolysis test, acute toxicity test, skin irritation test for comprehensive evaluation of the material biocompatibility and safety.

RESULTS AND CONCLUSION: The hydroxyapatite/nano-silver composite coating on the surface of titanium base exhibited obvious antibacterial activity in vitro against Staphylococcus aureus, in a silver dose-dependent manner. There was no pyrogen response, hemolytic reaction, acute toxic reaction and skin stimulus response in the animal experiments. There was also no difference in the biocompatibility of hydroxyapatite materials containing silver or not. These findings indicate that the hydroxyapatite/nano-silver composite coating on the surface of titanium base has good antibacterial activity for Staphylococcus aureus in vitro as well as excellent biocompatibility.

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

全文链接：

Key words: hydroxyapatites, silver, Staphylococcus aureus, titanium

CLC Number:

R318

Xian Ai-ming, Zhang Xiao-gang, Cao Li, Xu En-jie, Peng Li-bin, Hu Yang. In vitro antibacterial activity and biological safety of nano-silver composite coating on the surface of titanium base [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(52): 8387-8392.

Figures/Tables 2

2.1 纳米复合涂层材料体外抗菌试验结果 2.1.1 定性抑菌分析空白对照组和纯羟基磷灰石组生长有大量的细菌，大体观察二者之间菌落数量接近，许多菌落堆积在一起形成菌苔，菌体表面有一层白色的类似黏稠状的物质，表明细菌生长代谢旺盛，说明纯钛和不含银的纯羟基磷灰石不具有抗菌活性。1.0羟基磷灰石/银材料样品细菌菌落数减少最明显，只有零星的菌落分布在培养基上；0.5羟基磷灰石/银材料菌落数比1.0羟基磷灰石/银材料菌落数多(图1)。 2.1.2 定量抑菌分析在培养24 h以后，0.5羟基磷灰石/银和1.0羟基磷灰石/银两种材料对金黄色葡萄球菌的抗菌率都在95%以上，0.5羟基磷灰石/银和1.0羟基磷灰石/银两种材料抗菌率差异无显著性意义(P > 0.05)；而2组抗菌率均高于纯钛和羟基磷灰石组(P < 0.05)，说明羟基磷灰石/纳米银复合涂层有着很好的抗菌性能(表1)。 2.2 实验动物数量分析所有新西兰兔及大鼠均进入结果分析。 2.3 纳米复合涂层材料组织相容性试验结果 2.3.1 热源实验注射材料浸提液后，3组新西兰兔3 h内体温升高数均低于0.6 ℃。各组间差异无显著性意义(P > 0.05；表2)。 2.3.2 溶血实验 0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层3种材料和空白对照组试管溶液离心后，上清液均澄清，无色透明，红细胞沉积到管底，无溶血现象。阳性对照组蒸馏水试管中呈现玫瑰红色，红细胞完全破裂溶解。阳性对照组的吸光度与0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层3种材料组相比差异有显著性意义(P < 0.01)，而0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层三种材料组吸光度接近(P > 0.05)。0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层三种材料组与空白对照组的吸光度接近(P > 0.05)，且3组材料溶血率均小于非直接接触血液的医用生物材料溶血国际标准(5%)(表3)。 2.3.3 急性毒性实验注射材料浸提液后，0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层三种材料组和对照组均没有小鼠出现异常症状，活动、进食、排泄正常，未见步态不稳、惊厥、瘫痪以及呼吸抑制等毒性反应，体质量有增加趋势，观察期间无小鼠死亡，0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层三种材料组和对照组小鼠体质量变化无显著性意义(P > 0.05；表4)。 2.3.4 皮肤刺激试验皮内注射刺激试验观察1-72 h，0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层3种材料组注射点与对照侧注射点均无明显刺激反应。原发性皮肤刺激试验观察1-72 h，0.5羟基磷灰石/银、1.0羟基磷灰石/银、纯羟基磷灰石涂层3种材料组与阴性对照侧注射点均无局部红肿、渗出、溃烂等刺激反应。阳性对照侧皮肤1-24 h逐渐出现轻微红斑、水肿，48 h出现明显红斑及水肿，72 h出现焦痂。"

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