纳米丝强化型生物骨水泥治疗椎体骨质疏松骨折的细胞学机制考察

Cytological mechanism of nanowire enhanced biological bone cement on the treatment of vertebral osteoporosis fracture

目的分析RGD/纳米壳聚糖强化型磷酸钙骨水泥（CPC）与硫酸钙骨水泥（CSC）对体外间充质干细胞（MSC）粘附、增殖能力及促成骨分化能力的影响。方法取大鼠骨髓MSC,分别与CPC（CPC组）、壳聚糖强化型CPC（CPC-CH组）、RGD基团修饰的壳聚糖强化型CPC（CPC-CH-RGD组）、CSC（CSC组）、壳聚糖强化型CSC（CSC-CH组）以及RGD基团修饰的壳聚糖强化型CPC（CSC-CH-RGD组）体外接触式共培养4周。取培养液检测粘附分子细胞间黏附分子1（ICAM-1）及E选择素（E-selaction）含量、MSC细胞增殖率、凋亡率;并观察成骨细胞分化情况和矿化钙结节,测定成骨分化标记物OC、BMP的m RNA表达水平。结果使用CPC还是CSC作为基质骨水泥对MSC黏附、增殖、凋亡及促成骨分化功能无影响（P〉0.05）,与强化剂的配伍对MSC黏附、增殖、凋亡及促成骨分化功能有影响（P〈0.05）。其中CPC-CH-RGD组和CSC-CH-RGD组ICAM-1水平、E-selaction、MSC增殖率、成骨分化阳性率、钙结节数、OC m RNA表达量,BMP m RNA表达量均偏高,MSC凋亡率偏低,与其他各组相比,差异有统计学差异（P〈0.05）。结论 RGD/纳米壳聚糖强化型骨水泥能够促进体外MSC黏附、增殖及促成骨分化功能,可能为其改善骨质疏松患者预后的重要细胞学机制之一。

Objective To analyze the influence of calcium phosphate cement （CPC） and calcium sulfate cement （CSC） enhanced by RGD/CH itosan nanopartieles on the adhesion, proliferation and bone differentiation ability of Mesenchymal stem cells （MSC） in vitro. Methods The MSC of rat bone marrow were extracted and co-cultured with different kinds of bone cement, including CPC （CPC group）, ehitosan enhanced CPC （CPC-CH group）, RGD/CH itosan enhanced CPC （CPC-CH-RGDgroup）, CSC （CSC group）, chitosan enhanced CSC （CSC-CH group）, RGD/CH itosan enhanced CSC （CPC-CH-RGD group）. After 4 weeks, the co-cultural liquid was extracted to detect intercellular adhesion molecule lever, E-selaetion lever, MSC proliferation rate and apoptosis rate. Further more, osteoblast differentiation and mineralization calcium nodules were observed, mRNA expression of osteoblasts differentiation markers such as OC and BMP were detected too. Results Use CPC or CSC as the matrix of bone cement had no effect on MSC adhesion, proliferation, apoptosis and bone differentiation function （P〉O.05）, and the kinds of enhancer had effect on MSC adhesion, prolifer- ation, apoptosis, and bone differentiation function （P〈0.05）. The ICAM-1 level, E-selaetion lever, MSC proliferation rate, the osteogenesis differentiation rate, calcium nodules, OC mRNA expression and BMP mRNA expression quantity of both CPC-CH-RGD and CSC-CH-RGD group were higher than those of other groupsl and all the difference had statistical significance （P〈0.05）. Conclusion RGD/CH itosan nanopartieles enhanced bone cement can promote MSC adhesion, proliferation, and contribute to bone differentiation function in vitro, may be the one of the important cytology mechanisms of improving the prognosis of patients with osteoporosis.