纳米β磷酸三钙/水凝胶复合材料促进骨再生的实验研究  

Experimental study on promoting bone regeneration of nanoβtricalcium phosphate/hydrogel composite

作　　者：徐人杰[1,2] 马进进 周晓强 虞宵 颜勇卿 杨志杰 杨惠林 赛吉拉夫[2] 陈广祥 XU RenJie;MA Jinjin;ZHOU Xiaoqiang;YU Xiao;Yan Yongqing;YANG Zhijie;YANG Huilin;SAIJILAFU;CHEN Guangxiang(Department of Orthopaedics,Suzhou Municipal Hospital/The Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou,Jiangsu 215000,China;不详)

机构地区：[1]南京医科大学附属苏州医院/苏州市立医院骨科,江苏215000 [2]苏州大学骨科研究所,江苏215000 [3]中国科学院大学宁波华美医院骨科,浙江315000 [4]苏州鼎安科技有限公司,江苏215000

出　　处：《中国骨科临床与基础研究杂志》2021年第3期108-115,共8页Chinese Orthopaedic Journal of Clinical and Basic Research

基　　金：江苏省卫健委医学科研项目重点项目(K2019010);浙江省医药卫生科技面上项目(2019319060)。

摘　　要：目的观察纳米β磷酸三钙(TCP)/水凝胶复合材料对骨再生的促进作用。方法制作纳米β-TCP/甲基丙烯酸酰化明胶(GelMA)水凝胶复合材料。根据培养基的不同,分为空白对照组、GelMA组和纳米β-TCP/GelMA组,将大鼠骨髓间充质干细胞(BMSCs)与上述培养基共同培养。通过电镜观察、细胞增殖实验、碱性磷酸酶(ALP)染色,了解纳米β-TCP/GelMA复合材料的生物相容性及体外骨诱导活性。制作大鼠颅骨缺损模型,根据植入材料的不同,分为空白对照组、GelMA组和纳米β-TCP/GelMA组,术后4、8周取颅骨标本,通过Micro-CT扫描计算骨量分数和骨密度,对新骨形成情况进行组织学观察。结果大鼠BMSCs在材料表面培养48 h,扫描电镜观察发现其在GelMA组和纳米β-TCP/GelMA组表面良好贴附及铺展;细胞与材料共培养1、3、5 d,BMSCs数量随时间推移不断增加,3、5 d时纳米β-TCP/GelMA组OD值最高(P<0.05),7 d时与GelMA组相比,纳米β-TCP/GelMA组ALP表达明显增强。大鼠颅骨缺损区Micro-CT检查显示,纳米β-TCP/GelMA组骨量分数和骨密度高于空白对照组和GelMA组,差异具有统计学意义(P<0.05);组织学观察结果显示,与空白对照组、GelMA组相比,纳米β-TCP/GelMA组新生骨更丰富。结论纳米β-TCP/水凝胶复合材料具有良好的生物相容性,体外可促进大鼠BMSCs增殖和成骨分化,在大鼠颅骨缺损模型中具有良好的促进骨再生能力。Objective To observe the effect of nanoβtricalcium phosphate(TCP)/hydrogel composite in promoting bone regeneration.Methods NanoβTCP/methacrylate gelatin(GelMA)composite material was prepared.Three groups,blank control group,GelMA group and nanoβ-TCP/GelMA group were divided according to the different culture medium,and rat bone marrow mesenchymal stem cells(BMSCs)were cultured on the above media.The biocompatibility and in vitro bone induction activity of nanoβ-TCP/GelMA composites were investigated by electron microscope observation,cell proliferation assay and alkaline phosphatase(ALP)staining.The rat calvarial defect models were made and divided into blank control group,GelMA group and nanoβ-TCP/GelMA group according to the different implants.The calvarial specimens were taken for Micro-CT scan at 4 and 8 weeks postoperatively.The bone volume fraction(BV/TV)and bone mineral density(BMD)were calculated,and then the formation of new bone was observed histologically.Results After the rat BMSCs were cultured on the surface of the materials for 48 hours,scanning electron microscopy showed that the cells were attached and spread well on the surface of the materials in GelMA group and nanoβ-TCP/GelMA group.When cells and materials were co-cultured for 1,3 and 5 days,the number of BMSCs increased with time.When cells were co-cultured for 3 and 5 days,the OD value inβ-TCP/GelMA group was the highest among 3 groups(P<0.05).Compared with GelMA group,significant promotion of ALP expression was found inβ-TCP/GelMA group when the cells were co-cultured for 7 days.Micro-CT examination of the specimen showed that BV/TV and BMD in nanoβ-TCP/GelMA group were higher than those in blank control group and GelMA group(P<0.05).Histological observation showed that new bone was more abundant in nanoβ-TCP/GelMA group when compared with blank control group or GelMA group.Conclusion The nanoβ-TCP/hydrogel composite material has good biocompatibility,could promote the proliferation and osteogenic differentiation of rat BMSCs i

关 键 词：纳米复合物 磷酸钙类 水凝胶 生物相容性材料 骨代用品 骨髓细胞 干细胞 细胞 培养的 颅骨缺损 大鼠 骨生成 骨再生 

分 类 号：R318.17[医药卫生—生物医学工程] R329.471[医药卫生—基础医学]