Ruan Hongjiang, Liu Junjian, Fan Cunyi, Zheng Xuebin, Chen Yikai
Department of Orthopaedics, the Sixth People's Hospital, Shanghai Jiaotong University, Shanghai, 200233, P.R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2009 Feb;23(2):226-30.
To prepare silver-containing hydroxyapatite coating (hydroxyapatite/Ag, HA/Ag) and investigate its antibacterial property and biocompatibility in vitro.
Vacuum plasma spraying technique was adopted to prepare HA/Ag coating on titanium alloy substrate (3% Ag). After incubating the HA/Ag and the HA coating under staphylococcus aureus and pseudomonas aeruginosa suspensions of 2% tryptic soy broth (TBS) medium for 2, 4 and 7 days, respectively, the biofilm on the coatings was examined by confocal laser scanning microscope, and the bacterial density and viable bacterial percentage of bacterial biofilm were calculated. Meanwhile, the micro-morphology of bacterial biofilm was observed by SEM, the cytotoxicity was detected via MTT and the biocompatibility of biofilm was evaluated by acute haemolysis test.
Compared with HA coating, the bacterial biofilm's thickness on the surface of HA/Ag coating witnessed no significant difference at 2 days after culture (P > 0.05), but decreased obviously at 4 and 7 days after culture (P < 0.01). The bacterial density of the biofilm increased with time, but there was no significant difference between two coatings (P > 0.05) at 2, and 7 days after culture. The viable bacterial percentage of the biofilms on the surface of HA/Ag coating decreased obviously compared with that of HA coating at 2, 4 and 7 days after culture (P < 0.01). The MTT notified the cytotoxic grade of both coatings was zero. The acute haemolysis assay showed that the hemolytic rate of HA/Ag and HA coating was 0.19% and 0.12%, respectively.
With good biocompatibility, significant antibacterial property against staphylococcus aureus and pseudomonas aeruginosa, no obvious cytotoxicity and no erythrocyte destruction, the vacuum plasma sprayed HA/Ag coating is a promising candidate for the surface of orthopedic metal implants to improve their osseointegration and antibacterial property.
制备含银羟基磷灰石涂层(羟基磷灰石/银,HA/Ag),并体外研究其抗菌性能和生物相容性。
采用真空等离子喷涂技术在钛合金基底(含3%银)上制备HA/Ag涂层。将HA/Ag涂层和HA涂层分别置于2%胰蛋白胨大豆肉汤(TBS)培养基的金黄色葡萄球菌和铜绿假单胞菌悬液中孵育2、4和7天,之后用共聚焦激光扫描显微镜检测涂层上的生物膜,计算生物膜的细菌密度和活菌百分比。同时,通过扫描电子显微镜观察细菌生物膜的微观形态,采用MTT法检测细胞毒性,并通过急性溶血试验评估生物膜的生物相容性。
与HA涂层相比,培养2天后HA/Ag涂层表面细菌生物膜的厚度无显著差异(P>0.05),但培养4天和7天后明显降低(P<0.01)。生物膜的细菌密度随时间增加,但培养2天和7天后两种涂层之间无显著差异(P>0.05)。培养2、4和7天后,HA/Ag涂层表面生物膜的活菌百分比与HA涂层相比明显降低(P<0.01)。MTT法显示两种涂层的细胞毒性等级均为零。急性溶血试验表明,HA/Ag涂层和HA涂层的溶血率分别为0.19%和0.12%。
真空等离子喷涂的HA/Ag涂层具有良好的生物相容性,对金黄色葡萄球菌和铜绿假单胞菌有显著抗菌性能,无明显细胞毒性且无红细胞破坏,是改善骨科金属植入物表面骨整合和抗菌性能的有前景的候选材料。