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目的研究不同锶(Sr)掺杂量的羟基磷灰石(HA)的生物相容性及生物学活性。方法以Sr含量分别为0%、1%、5%、10%、20%的5组Sr掺杂的HA(Sr-HA)材料作为试样,检测各组试样浸提液中Ca2+、Sr2+浓度;通过溶血试验检测5组材料的体外溶血性;将L929成纤维细胞在5组材料浸提液中接种培养2、7d,通过MTT法检测5组材料体外细胞毒性,用倒置相差显微镜观察细胞形态;将MG63成骨细胞接种到5种材料表面,采用MTT方法检测培养1、4、12h后成骨细胞在材料表面的黏附活性和1、4、7d时细胞在材料表面的增殖活性,并对培养1、3、5d时成骨细胞碱性磷酸酶(ALP)活性进行检测。结果在5组Sr-HA材料浸提液中,Ca2+浓度以5%Sr含量组最高、1%Sr含量组最低,Sr2+浓度以20%Sr含量组最高、0%Sr含量组最低;各组Sr-HA溶血率均低于5%,显微镜观察5组材料表面接种培养的L929细胞形态与阴性对照组相似,MTT法检测5组材料体外细胞毒性评级为0级、1级;MG63成骨细胞在材料表面培养4、12h,5%Sr含量、10%Sr含量、20%Sr含量3组材料表面细胞黏附数量明显高于0%Sr含量、1%Sr含量组(P<0.05,P<0.01);MG63成骨细胞在材料表面培养4、7d,5%Sr含量、10%Sr含量、20%Sr含量3组材料表面细胞增殖数量明显高于0%Sr含量、1%Sr含量组(P<0.05);MG63成骨细胞在5组材料表面接种培养3d时,5%Sr含量、10%Sr含量2组ALP活性高于0%Sr含量、1%Sr含量2组(P<0.05),5d时5%Sr含量、10%Sr含量、20%Sr含量3组ALP活性高于0%Sr含量、1%Sr含量2组。结论不同Sr含量的Sr-HA具有良好的生物相容性;一定浓度的Sr掺杂能够有效提高HA的生物学活性,Sr含量为5%、10%时Sr-HA生物学效果最佳。
Objective To study the biocompatibility and biological activity of hydroxyapatite (HA) with different strontium (Sr) content. Methods Five Sr-doped HA (Sr-HA) materials with Sr content of 0, 1, 5, 10 and 20% were used as samples to detect Ca2 + and Sr2 + The hemolytic activity of five groups of materials was tested by hemolysis test. L929 fibroblasts were inoculated for 2 and 7 days in 5 groups of material extracts. The in vitro cytotoxicity of 5 materials were detected by MTT assay. The cells were observed by inverted phase contrast microscope MG63 osteoblasts were inoculated on the surface of five kinds of materials. MTT assay was used to detect the adhesion activity of osteoblasts on the surface of the material after 1, 4 and 12 h culture and the proliferation activity of the cells on the surface of the material at 1, 4 and 7 days Osteoblasts alkaline phosphatase (ALP) activity was detected at 1, 3 and 5 days after culture. Results In the five Sr-HA extracts, Ca2 + concentration was the highest in 5% Sr, the lowest in 1% Sr, the highest in Sr2 + and the lowest in 0% Sr. -HA hemolysis rates were less than 5%. The morphological changes of L929 cells inoculated on the surface of 5 groups were similar to those of the negative control group. The MTT assay was used to evaluate the cytotoxicity of 5 materials in vitro. The number of cell adhesion on the surface of the material was significantly higher than that of 0% Sr and 1% Sr (P <0.05, P <0.01) after 4,12h, 5% Sr, 10% Sr and 20% MG63 osteoblasts cultured on the surface of the material for 4, 7 days, 5% Sr content, 10% Sr content and 20% Sr content were significantly higher than 0% Sr and 1% Sr (P < 0.05). MG63 osteoblasts were cultured on the surface of 5 groups for 3 days. The ALP activity of 5% Sr and 10% Sr was higher than that of 0% Sr and 1% Sr (P <0.05) When 5% Sr content, 10% Sr content and 20% Sr content were higher than 0% Sr content, 1% Sr content was 2 groups. Conclusion Sr-HA with different content of Sr has good biocompatibility. Sr concentration of 5% Sr can enhance the biological activity of Sr effectively. Sr-HA with 10% concentration has the best biological effect.