摘要
背景:偏磷酸钙具有优异的细胞相容性能和降解性能及细胞亲和性,人骨髓间充质干细胞可以在多孔偏磷酸钙孔洞内生长和增殖,但有关偏磷酸钙纳米粒子的研究较少。目的:制备纳米级偏磷酸钙微粒,通过流式细胞术快速检测不同浓度纳米级偏磷酸钙微粒对人骨髓间充质干细胞凋亡的影响。方法:采用湿法球磨法制备偏磷酸钙纳米粒子,通过扫描电镜和透射电镜观察纳米粒子的形貌,通过X射线衍射分析确定纳米粒子的晶体结构。将偏磷酸钙纳米粒子混入CYAGON OricellTM基础培养基,使得偏磷酸钙纳米粒子的质量浓度分别为10,1,0.1 mg/L,将其与人骨髓间充质干细胞共培养7 d,通过流式细胞术分析偏磷酸钙纳米粒子质量浓度与细胞凋亡的关系。结果与结论:采用湿法球磨法成功制备了偏磷酸钙纳米粒子,直径为10-30 nm,粒径分布较均匀,分散性较好,但晶体形状不规则;X射线衍射分析晶相检测其主晶相为β-Ca(PO3)2晶体。10 mg/L质量浓度组细胞G0/G1和G2/M比例高于1,0.1 mg/L质量浓度组(P<0.01);10 mg/L质量浓度组细胞早期、中晚期、总细胞凋亡率高于1,0.1 mg/L质量浓度组(P<0.01);说明偏磷酸钙纳米粒子对人骨髓间充质干细胞的增殖有影响,当其质量浓度从1 mg/L增加至10 mg/L后,细胞凋亡率显著增加。
BACKGROUND：Calcium metaphosphate has excel ent biocompatibility, degradability, and cel affinity. Human bone marrow mesenchymal stem cel s can grow and proliferate in the pores of the porous calcium metaphosphate, but less is known about calcium metaphosphate nanoparticles.
OBJECTIVE：To prepare calcium metaphosphate nanoparticles, and to analyze the effect of calcium metaphosphate nanoparticles at different concentrations on apoptosis of human bone marrow mesenchymal stem cel s by flow cytometry.
METHODS：The calcium metaphosphate nanoparticles were prepared by wet bal mil ing. Scanning electron microscopy and transmission electron microscopy were used to observe the morphology of the calcium metaphosphate nanoparticles, and the crystal structure of nanoparticles was analyzed by X-ray diffraction. Calcium metaphosphate nanoparticles were mixed in the CYAGON Oricel TM basal medium, and the concentrations of calcium metaphosphate nanoparticles in the medium were 10, 1, 0.1 mg/L. Human bone marrow mesenchymal stem cel s were cultured for 7 days in the above-mentioned media, and apoptosis of human bone marrow mesenchymal stem cel s was analyzed by flow cytometry.
RESULTS AND CONCLUSION：Calcium metaphosphate nanoparticles were successful y prepared by wet bal mil ing, irregular in shape, and the mean diameter was 10-30 nm. X-ray diffraction results showed the crystal structure of nonaparticles was mainlyβ-Ca（PO3）2. The cel ratio of G0/G1 phase and G2/M phase in 10 mg/L group was obviously higher than that in 1, 0.1 mg/L groups （P〈0.01）. The cel apoptosis rates during the early, middle, late stages in 10 mg/L group were obviously higher than those in 1, 0.1 mg/L groups （P〈0.01）, and the total cel apoptosis was also significantly increased in 10 mg/L group （P〈0.01）. These findings indicate that human bone marrow mesenchymal stem cel s proliferation can be inhibited by calcium metaphosphate nanoparticles, and apoptosis rate is increased significantly when the concentration of calcium metaphosphate nanoparticles increases from 1 mg/L to 10 mg/L.
出处
《中国组织工程研究》
CAS
CSCD
2013年第38期6797-6803,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金资助项目(30901468)~~
