锶掺杂改性羟基磷灰石的生物学性能研究

目的：研究不同锶掺杂量的羟基磷灰石Sr-HA的生物相容性及生物学活性。方法：合成0%(A)、1%(B)、5%(C)、10%(D)、20%(E)掺锶含量不同的五组Sr-HA,检测各组试样浸提液中钙、锶离子浓度,通过溶血试验检测了五组材料体外溶血性。L929成纤维细胞材料表面接种培养2、5天,通过倒置相差显微镜观察细胞形态MTT法检测相对增值度测试五组材料体外细胞毒性。将MG63成骨细胞接种到五种材料表面,采用MTT方法检测1、4、12小时成骨细胞在材料表面的附着和1、4、7 d时细胞在材料表面的增殖情况;并对第1、3、5天成骨细胞ALP活性进行检测。结果：100%浸提液五组材料中Ca₂₊浓度5%(C)最高,1%(B)最低;Sr₂₊浓度,20%(E)最高,0%(A)组最低。各组掺锶羟基磷灰石组的溶血率均低于5%。显微镜观察L929细胞的形态与阴性对照组比较相似。MTT检测相对增殖度5组材料总体毒性评级为0级、1级。MG63成骨细胞在材料表面培养4、12小时, 5%(C)、10%(D)、20%(E)三组表面细胞粘附数量明显高于0%(A)、1%(B)组 (P<0.05,P<0.01);12小时：1%(B)细胞粘附数量高于0%(A)组(P<0.05)。MG63成骨细胞在材料表面培养4、7天,5%(C)、10%(D)、20%(E)三组表面细胞增殖数量明显高于0%(A)、1%(B)组 (P<0.05,)。ALP活性检查显示：3天时5%(C)、10%(D)两组ALP活性高于其它三组(P<0.05,)。5天时5%(C)、10%(D)、20%(E)三组ALP活性高于0%(A)、1%(B)两组(P<0.05,)。结论：不同锶含量掺杂的羟基磷灰石具有良好地生物相容性,一定浓度的锶离子掺杂能够有效的提高羟基磷灰石的生物学活性,我们认为5%(C)、10%(D)组掺杂Sr-HA生物学效果最佳。

Study on the biological properties of strontium doped hydroxyapatite

Objective: To study the biocompatibility and biological activity of hydroxyapatite with different strontium content. Methods: five groups of Sr-HA with 0% (A),1% (B), 5% (C), 10% (D), and,20% (E) respectively, were prepared to detect the concentration of calcium and strontium ions in each group. L929 fibroblasts were cultured for 2 days and 5 days, cell morphology was observed by inverted phase contrast microscope and the relative value of the five groups was detected by MTT assay. The MG63 bone cells were seeded into five kinds of material surface for1, 4, 12 hours, and 1, 4 and 7 days,MTT method was used to determine proliferation and adhesion of 0steoblasts at different time points ; and ALP activity of osteoblast were detected at1,3,5day.Results: In the five groups, the concentration of calcium was highest in 5%(C) group, and the lowest in 1% (B) group, the concentration of strontium was highest in20% (E) group, and the lowest in 0% (A) group.The hemolytic rate of strontium substituted hydroxyapatite was lower than 5%. The morphology of L929 cells was similar to that of the negative control group. The overall toxicity of the groups was detected with the MTT assay, and the toxicity of the 5 groups was 0 and 1. MG63 osteoblasts were cultured in for 4 and 12 hours, The number of cell surface adhesion in 5% (C), 10% (D), 20%(E) was significantly higher than that of A and B(P<0.05 P<0.01). At 12 hours, the number of cell adhesion in 1% (B) group was significantly higher than that in 0%(A) group(P<0.05).MG63 osteoblasts were cultured on the surface of the material for 4 and7days, days, the number of cell proliferation in5% (C), 10% (D) and20% (E), was significantly higher than that in 0% (A) group and 1% (B) group (P<0.05). ALP activity of 5% (C) group and 10% (D) group were higher than those in the other three groups at third day (P<0.05,). ALP activity of 5% (C), 10% (D),and20% (E) were higher than 0% (A) group and1% (B) group at fifth day (P<0.05). Conclusion: the hydroxyapatite with different strontium content is a good biocompatibility, and a certain concentration of strontium ions can effectively improve the biological activity of hydroxyapatite. We think that 5% (C) group and 10% (D) group are the best.