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锶掺杂改性羟基磷灰石的生物学性能研究
高建勇,田刚,朱强,王少海,汪大林*
0
(第二军医大学长海医院口腔科, 上海 200433
*通信作者)
摘要:
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、7 d,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、12 h后成骨细胞在材料表面的黏附活性和1、4、7 d时细胞在材料表面的增殖活性,并对培养1、3、5 d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、12 h, 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、7 d,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3 d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),5 d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。
关键词:    羟基磷灰石类  成骨细胞  生物学活性  生物相容性
DOI:10.16781/j.0258-879x.2016.02.0138
投稿时间:2015-11-17修订日期:2016-01-09
基金项目:军事口腔医学国家重点实验室开放课题(2014KB07),上海市卫生局局级课题面上项目(20133124),第二军医大学长海医院"1255"基金项目(CH125541800).
Study on the biological properties of strontium doped hydroxyapatite
GAO Jian-yong,TIAN Gang,ZHU Qiang,WANG Shao-hai,WANG Da-lin*
(Department of Stomatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
*Corresponding author)
Abstract:
Objective To study the biocompatibility and biological activity of hydroxyapatite (HA) with different strontium (Sr) contents. Methods The Sr doped HA (Sr-HA) materials containing 0%, 1%, 5%, 10%, and 20% Sr were prepared and the concentrations of Ca2+ and Sr2+ in the leaching liquor of them were determined in each group. Hemolysis tests were performed in vitro for the 5 materials. L929 fibroblasts were seeded and cultured with leaching liquor of materials for 2 d and 7 d; cell morphology was observed under microscope and the in vitro cytotoxicity of the five materials was detected by MTT assay. The MG63 bone cells were seeded on the surface of the 5 materials; then cell adhesion was observed at 1, 4, 12 h of culture, and the proliferation of cells was observed on day 1, 4 and 7 of culture by MTT method; and alkaline phosphatase (ALP) activity of the osteoblasts was detected on day 1, 3, and 5. Results The highest Ca2+concentration was found in the leaching liquor of 5% Sr group, and the lowest was found in the 1% Sr group; the highest Sr2+concentration was found in the leaching liquor of 20% Sr group, and the lowest was found in 0% Sr group. The hemolytic rates of Sr-HA materials were lower than 5% in all the five Sr-HA groups. The morphology of L929 fibroblasts in the five Sr-HA groups was similar to that of the negative control group; and MTT assay showed that the overall cytotoxicity of the five Sr-HA groups was grade 0 or 1. After 4 and 12 h culture, the number of adhered MG63 osteoblasts in 5%, 10%, and 20% Sr groups were significantly more than those in 0% and 1% Sr groups (P<0.05, P<0.01). On day 4 and 7 of culture, the proliferation rates of MG63 osteoblasts in 5%, 10% and 20% Sr groups were significantly higher than those in 0% and 1% Sr groups (P<0.05). The ALP activity of MG63 osteoblasts in 5% and 10% Sr groups was significantly higher than that in 0% and 1% Sr groups on day 3 of culture (P<0.05); and that in 5%, 10%, and 20% Sr groups was higher than that in 0% and 1% Sr groups on day 5 of culture. Conclusion Sr-HA material with different Sr contents has good biocompatibility; the combination with certain concentration of Sr can effectively improve the biological activity of HA, with the best activity seen when Sr ratio is 5% or 10%.
Key words:  strontium  hydroxyapatites  osteoblasts  bioactivity  biocompatibility