目的 探讨纯钛表面负载无定形多聚磷酸钙(Ca-polyP)涂层对成骨分化的影响.方法 取TA2级纯钛片,表面构建Ca-polyP涂层,采用场发射扫描电子显微镜、X射线能量色散谱仪、X射线衍射仪、傅立叶变换红外光谱仪和接触角测量仪对材料进行表征.将小鼠胚胎成骨前体细胞接种于纯钛片(对照组)以及负载Ca-polyP涂层的钛片(实验组)上进行细胞培养,采用CCK-8法测定细胞的增殖活性,通过碱性磷酸酶活性测定以及碱性磷酸酶染色评估两组钛片表面细胞的成骨分化能力.结果 多种物理表征方法检测结果证明20链长Ca-polyP涂层成功负载在钛片表面.CCK-8检测结果显示,两组钛片均无细胞毒性,且共培养7 d时实验组钛片表面的细胞增殖活性显著高于对照组,差异具有统计学意义(F=1 375.183,P＜0.001).随着共培养时间的延长,两组钛片表面细胞的碱性磷酸酶活性均逐渐增高,共培养7 d时实验组钛片表面细胞的碱性磷酸酶活性显著高于对照组,差异具有统计学意义(F=41.141,P＜0.01).碱性磷酸酶染色后荧光倒置显微镜下观察,实验组钛片上的蓝紫色深染结节明显多于对照组.结论 与纯钛相比,钛片表面负载Ca-polyP涂层可以显著增强小鼠胚胎成骨前体细胞的增殖和成骨分化能力.
Objective To investigate the effect of pure titanium surface-loaded amorphous calcium polyphosphate(Ca-polyP)coating on osteogenic differentiation.Methods TA2 titanium sheets were taken,coated with Ca-polyP on the surface,and characterized by field emission scanning electron microscopy,X-ray energy dispersive spectroscopy,X-ray diffractometry,Fou-rier transform infrared spectroscopy,and contact angle measurement.Mouse embryo osteoblast precursor cells were inoculated on pure titanium sheets(control group)and titanium sheets loaded with Ca-polyP coating(experimental group)for cell culture.The proliferation activity of cells was measured by CCK-8 assay.The osteogenic differentiation ability of cells on the surface of titanium sheets in both groups was assessed by alkaline phosphatase activity assay and alkaline phosphatase staining.Results Successful loading of 20-chain-length Ca-polyP coating on titanium sheet surface was demonstrated by various physical characterization me-thods.The results of CCK-8 assay showed that titanium sheets in both groups were non-cytotoxic,and the cell proliferation activity on the surface of titanium sheets in the experimental group was significantly higher than that in the control group after 7 days of co-culture(F=1 375.183,P＜0.001).With the extension of co-culture time,the alkaline phosphatase activity of cells on the surface of titanium sheets gradually increased in both groups,and the alkaline phosphatase activity of cells on the surface of titanium sheets in the experimental group was significantly higher than that in the control group after 7 days of co-culture(F=41.141,P＜0.01).Al-kaline phosphatase staining followed by inverted fluorescence mi-croscopy showed that there were significantly more blue-purple dark-ly stained nodules on titanium sheets in the experimental group than in the control group.Conclusion Titanium sheets loaded with Ca-polyP coating on the surface significantly enhances the proliferation and osteogenic differentiation of mouse embryo osteoblast precursor cells compared with pure titanium sheets.
单烁;周欢;陈胜男;魏子然;杨磊;于腾波
青岛大学附属医院运动医学科,山东青岛 266100||青岛大学运动医学与健康研究院健康科学与工程研究中心/河北省生物医学材料与智能诊疗重点实验室/生命科学与健康工程学院/河北工业大学青岛大学附属医院小儿外科青岛大学运动医学与健康研究院||康复大学青岛医院(青岛市立医院)骨科
基础医学
钛;磷酸钙类;小鼠胚胎干细胞;成骨分化
titanium;calcium phosphates;mouse embryonic stem cells;osteogenic differentiation
《青岛大学学报（医学版）》 2024 (001)
6-11 / 6
国家自然科学基金杰出青年基金(82025025);国家自然科学基金青年科学基金(31802022);山东省重点研发计划项目(2021SFGC0502);河北省自然基金绿色通道项目(C2021-202003);河北省自然基金创新研究群体项目(H2022202007);天津市多元投入基金项目面上项目(21JCYBJC01030);国家骨科与运动康复临床医学研究中心创新基金项目(2021-NCRC-CXJJ-Z-H-17)
10.11712/jms.2096-5532.2024.60.014
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