医用新型Mg-Li-Ca合金材料体外生物相容性及生物活性评价

In vitro biocompatibility and bioactivity of a new medical Mg-Li-Ca alloy

背景:新型Mg-Li-Ca合金是否具有生物相容性和生物活性,目前还未被证实。目的:通过体外实验评价新型医用Mg-Li-Ca合金的组织相容性及生物活性,初步探讨其作为医用植入材料的可行性。方法:分别采用Mg-Li-Ca合金浸提液、纯Mg浸提液、AZ31B合金浸提液与α-MEM培养液培养第3代小鼠胚胎成骨细胞,培养1,3,5 d,采用MTT法检测细胞A值并计算相对增殖率;培养5,7 d,使用碱性磷酸酶试剂盒检测细胞碱性磷酸酶活性。将第3代小鼠胚胎成骨细胞与Mg-Li-Ca合金、纯Mg、AZ31B合金分别共培养于24孔板,培养1 d后扫描电镜观察材料表面黏附及增殖情况。结果与结论:Mg-Li-Ca合金、纯Mg和AZ31合金对成骨细胞的细胞毒性为Ⅰ级,无细胞毒性,且Mg-Li-Ca合金对细胞毒性明显小于纯Mg和AZ31B合金,对成骨细胞生长增殖无显著影响,呈现出良好的生物相容性。Mg-Li-Ca合金和纯Mg对成骨细胞正常合成碱性磷酸酶无影响,具有良好的生物活性;AZ31B对成骨细胞正常合成碱性磷酸酶有显著影响。成骨细胞可在Mg-Li-Ca合金、纯Mg和AZ31合金表面正常黏附生长。表明新型Mg-Li-Ca合金有望成为新型骨科植入材料。

BACKGROUND： Whether the new type of Mg-Li-Ca alloy has good biocompatibility and bioactivity or not has not been confirmed. OBJECTIVE： To evaluate the in vitro biocompatibility and bioactivity of a new medical Mg-Li-Ca alloy, and to find the possibility of this Mg-Li-Ca alloy as a new type of implant material. METHODS： The MC3T3-e1 （mouse osteoblasts） was used to be the test cells, then cultured and subcultured in vitro at passage 3. The cells were cultured and developed by leaching liquors of Mg-Li-Ca alloy, pure magnesium, and AZ31B alloy, and α-minimum essential medium, respectively. MTT assay was used to detect the absorbance values at days 1, 3, 5 of culture, and then the relative growth rate was calculated. Alkaline Phosphatase Colorimetric Assay Kit was used to check the activity of alkaline phosphatase at days 5 and 7 of culture. And the MC3T3-e1 cells were cocultured with Mg-Li-Ca alloy, pure magnesium, and AZ31B alloy, respectively; and at 1 day of coculture, cell adhesion and proliferation was observed under scanning electron microscopy. RESULTS AND CONCLUSION： Mg-Li-Ca alloy, pure magnesium, and AZ31B alloy were non-cytotoxic to osteoblasts, and the cytotoxicity of Mg-Li-Ca alloy was lower than that of pure magnesium and AZ31B alloy, indicating Mg-Li-Ca alloy had no influence on osteoblast proliferation and had good biocompatibility. The alkaline phoaphatase activity was not influenced by Mg-Li-Ca alloy and pure magnesium that had good bioactivity. AZ31B had a significant impact on the normal synthesis of alkaline phosphatase in osteoblasts. The MC3T3-e1 cells can adhere and grow normally on the surface of Mg-Li-Ca alloy, pure magnesium, and AZ31B alloy. These findings indicate that the new type of mg-Li-Ca alloy is expected to be a new bone implant material.