钙磷涂层镁合金对兔骨骼肌细胞生物学行为的影响

The influence of magnesium alloy with calcium phosphate coating on the biological behavior of rabbit skeletal muscle cells

目的探讨钙磷涂层镁合金对兔骨骼肌细胞生物学行为的影响。方法将兔骨骼肌细胞分别接种于钛合金(钛合金组)、镁合金(AZ31B组)及钙磷涂层镁合金(CaP-AZ31B组)试件上培养2、6、24h,计算细胞黏附率,扫描电镜观察细胞黏附形态;采用AZ31B及CaP-AZ31B浸提液培养兔骨骼肌细胞1、3、5、7d,同时以不加合金浸提液的细胞作为对照,倒置显微镜观察细胞形态,CCK-8法检测细胞增殖活力,Calcein-AM和EthD-1双染法观察培养4d后细胞存活情况,并测定细胞内总蛋白。结果随着培养时间的延长,3组细胞黏附率逐渐增加(P<0.05);培养2、6、24h时,3组细胞黏附率比较,差异有统计学意义(P<0.05)。浸提液培养1、3、5、7d,AZ31B组细胞相对增殖率低于对照组(P<0.05),CaP-AZ31B组高于AZ31B组(P<0.05);AZ31B组毒性评级为2级,CaP-AZ31B组毒性评级为1级;对照组、CaP-AZ31B组细胞活性较好,无明显细胞红染,AZ31B组部分细胞红染,但绝大多数细胞仍具有活性;3组细胞内总蛋白量呈时间依赖性增加(P<0.05),各时相点3组细胞内总蛋白量比较,差异有统计学意义(P<0.05),从高到低依次为对照组、CaP-AZ31组、AZ31B组。结论钙磷涂层镁合金对于兔骨骼肌细胞具着良好的生物相容性。

Objective To investigate the influence of magnesium alloy with calcium phosphate coating on the biological behavior of rabbit skeletal muscle cells. Methods Rabbit skeletal muscle cells were inoculated on titanium alloy, magnesium alloy （AZ31B） and magnesium alloy with calcium phosphate coating （CaP-AZ31B）,＆nbsp;and were cultured for 2, 6, 24 h. Cell adhesion rates were calculated, and cell adhesion morphology were observed by scanning electron microscope. In AZ31B and CaP-AZ31B leaching liquids, rabbit skeletal muscle cell were cultured for 1, 3, 5, 7 d, and cultured cells without alloy extracts were used as controls. Cell morphology was observed by inverted microscope, cell proliferation was detected by CCK-8 method, cell survival at 4 d after culture were evaluated by Calcein-AM and EthD-1 double staining, and the total cellular protein were determined in those 3 groups. Results Cell adhesion rates in 3 groups increased time-dependently （P 〈0.05）;At 2, 6, 24 h, the differences of cell adhesion rate among 3 groups had statistically significant （P 〈0.05）. Cell relative growth rate in AZ31B group was lower than that of control group at 1, 3, 5, 7 d after the culture with alloy extracts （P 〈0.05）, while the rate in CaP-AZ31B group was higher than that of AZ31B group at each timepoint （P 〈0.05）; Toxicity in AZ31B group was rated as 2 grade, and in CaP-AZ31B group rated as 1 grade; Cell activities were good in CaP-AZ31B group as well as the control group without obvious red-staining cells, while part of the cells were red-stained in AZ31B group with most of them still active; Intracellular total protein content in 3 groups increased in a time-dependent manner （P 〈0.05）, the differences of total protein content among 3 groups at each timepoint were statistically significant （P 〈0.05）, and the ranking of the total protein content from high to low was control group, CaP-AZ31 group, AZ31B group accordingly. Conclusion Magnesium alloy with calcium phosphate coating has a good biocompatibility to rabbit skeletal muscle cells.