壳聚糖-阿仑膦酸钠对大鼠成骨细胞体外增殖和分化的影响  

Effect of chitosan combined with alendronate on proliferation and differentiation of rat osteoblasts in vitro

作　　者：张晓荣[1] 孙国栋[1] 邵晓虹 李志忠[1] ZHANG Xiao-rong;SUN Guo-dong;SHAO Xiao-hong;LI Zhi-zhong(Orthopaedic Department,Affiliated Hospital,Jinan University,Guangzhou 510630,China;Department of Material Science,College of Engineering,Jinan University,Guangzhou 510630,China)

机构地区：[1]暨南大学附属第一医院骨科,广州510630 [2]暨南大学理工学院材料科学系,广州510630

出　　处：《中国矫形外科杂志》2018年第18期1697-1703,共7页Orthopedic Journal of China

基　　金：广东省中医药局科研项目(编号:20173018)

摘　　要：[目的]观察三种壳聚糖-阿仑膦酸钠(Cs/ALN)复合材料对体外成骨细胞增殖与分化的影响。[方法]用酶消化法分离培养新生SD大鼠颅盖骨的成骨细胞。制备三种比例的Cs/ALN复合材料及不含ALN的单纯Cs支架材料。将细胞分别与4种材料复合培养,Transwell法观察材料对细胞的趋化作用;流式细胞术检测材料对细胞生长周期的影响;酶标仪法、ELISA法分别检测细胞培养上清液中碱性磷酸酶(ALP)及骨钙素(BGP)的含量。[结果]流式细胞术细胞周期检测结果显示10∶1 Cs/ALN复合材料对成骨细胞增殖的促进作用最强。细胞培养上清液ALP及BGP含的测定显示10∶1 Cs/ALN复合材料对成骨细胞分化的促进作用亦最强。5∶1 Cs/ALN和20∶1 Cs/ALN两种复合材料对成骨细胞的增殖和分化也有一定的促进作用;而Cs支架材料则对成骨细胞的增殖和分化无明显促进作用;Transwell试验结果显示,5∶1 Cs/ALN复合材料对成骨细胞的趋化作用最强。[结论]10∶1 Cs/ALN复合材料具有明显促进成骨细胞增殖、分化及一定的趋化作用。[Objective] To explore the effect of chitosan（Cs） combined with alendronate（ALN） in different proportions on proliferation and differentiation of rat osteoblasts in vitro. [Methods] The osteoblasts were isolated from neonate SD rats by being digested with collagenase and trypsin. Cs/ALN composites in ratio of 5：1, 10：1 and 20：1 were made as scaffolds for cell culture, additionally, a pure Cs scaffold without ALN also made. The osteoblasts in the third generation were co-cultured with the above four scaffolds. The effect of the four materials on the chemotaxis of the cells was observed by transwell assay, while the proliferation and cellcycle was assessed by flow cytometry, and the osteogenic differentiation was identified by determining alkaline phosphatase（ALP） and bone glaprotein（BGP） in the cell culture supernatant, [Results] The Cs/ALN in 10：1 proved the most powerful effect on proliferation of osteoblasts among the four materials determined by flow cytometry. In addition, the Cs/ALN in 10：1 presented the most powerful effect on osteogenic differentiation of the cell by detecting the ALP and BPG in the cell culture supernatant. Although both Cs/ALN in 5：1 and Cs/ALN in 20：1 also showed improving effect on proliferation and osteogenic differentiation of the cells in some extent, the pure Cs had no significant role on proliferation and differentiation of the cells. However, the Cs/ALN in 5：1 revealed the most powerful effect chemotaxis of the cells by the transwell assay. [Conclusions] The chitosan/alendronate composite in ratio of 10：1 material has significant improving effect on proliferation and differentiation, and some role on chemotaxis of osteoblasts.

关 键 词：SD大鼠 成骨细胞 壳聚糖 阿仑膦酸钠 复合材料 

分 类 号：R318[医药卫生—生物医学工程]