SLM含铜钴基合金的抗菌性能及细胞相容性的初步探索

目的 以SLM-CoCrW为对照，评价SLM-CoCrCu的抗菌性及生物相容性，为SLM-CoCrCu的潜在临床应用提供理论依据。方法 选择金黄色葡萄球菌为研究对象，采用平板法验证合金的抗菌性能。选择MC3T3-E1细胞为研究对象，采用CCK-8检测法评价合金对于细胞增殖的影响和细胞毒性等级。采用细胞骨架荧光染色标记法评价细胞在合金表面的粘附及生长，结合增殖率，反映合金的细胞毒性。采用流式细胞术评价材料对细胞早期凋亡的影响。结果 SLM-CoCrCu对金黄色葡球菌的生长有明显抑制作用，对金黄色葡萄球菌抗菌率可达98%，与SLM-CoCrW比较有显著性差异（P<0.01）。随培养时间的延长，SLM-CoCrCu细胞数量逐步增长，表现出对细胞增殖的促进作用。SLM-CoCrCu和SLM-CoCrW在与细胞共培养1、3、 7 d后，细胞增殖率有显著差异性（P=0.01）。共培养1 d后，不含铜组细胞增殖率优于含铜组；共培养3 d及7 d后，含铜组对细胞增殖影响明显优于不含铜组。两者对于细胞粘附及形态上的影响无明显差异。SLM-CoCrCu和SLM-CoCrW对于细胞凋亡的影响无明显差异。结论 SLM-CoCrCu相较于SLM-CoCrW有良好的抗菌性能。同时，SLM-CoCrCu具有良好的生物相容性，对细胞增殖、粘附及形态都无明显影响。

Preliminary Exploration on Antibacterial Property and Biocompatibility of SLM Copper-Cobalt Alloys

The work aims to use SLM-CoCrW as a control to evaluate the antimicrobial and biocompatibility of SLM-CoCrCu, and then provide a theoretical basis for the potential clinical application of SLM-CoCrCu. Staphylococcus aureus was selected as the research object, and the antimicrobial properties of the alloy were tested by plate method. MC3T3-E1 cells were selected as the research object. CCK-8 assay was used to evaluate the effect of alloy on cell proliferation and cytotoxicity. The adhesion and growth of cells on the alloy surface were evaluated by cytoskeleton fluorescence staining, and the cytotoxicity of the alloy was reflected by the proliferation rate. Flow cytometry was used to evaluate the effect of materials on early cell apoptosis. SLM-CoCrCu obviously inhibited the growth of staphylococcus aureus, and the antimicrobial rate of SLM-CoCrCu was 98%. There was a significant difference between SLM-CoCrCu and SLM-CoCrW (P<0.01). With the prolongation of culture time, the number of SLM-CoCrCu cells increased gradually, and promoted cell proliferation. SLM-CoCrCu and SLM-CoCrW co-cultured with cells for 1, 3 and 7 days showed significant difference in cell proliferation rate (P=0.01). After co-culture for 1 day, the cell proliferation rate of copper-free group was better than that of copper-free group. After co-culture for 3 days and 7 days, the effect of copper-containing group on cell proliferation was significantly better than that of copper-free group. There was no significant difference in cell adhesion and morphology between the two groups. There was no significant difference between SLM-CoCrCu and SLM-CoCrW on apoptosis. Compared with SLM-CoCrW, SLM-CoCrCu has better antimicrobial properties. At the same time, SLM-CoCrCu has good biocompatibility and has no significant effect on cell proliferation, adhesion and morphology.