摘要
目的:探讨负载二甲基乙二酰甘氨酸(dimethyloxalyl glycine,DMOG)的光交联明胶(gelatin methacryloyl,GelMA)水凝胶的促成骨分化作用以及对大鼠颅骨骨缺损的修复效果。方法:制备GelMA-DMOG水凝胶后,扫描电镜下观察其表面特征,体外检测其降解时间;将小鼠成骨前体细胞MC3T3-E1培养于GelMA和GelMA-DMOG水凝胶表面,鬼笔环肽染色荧光观察细胞骨架形态,CCK-8法检测其细胞毒性;成骨诱导后,通过碱性磷酸酶(alkaline phosphatase,ALP)染色、茜素红染色和qRTPCR检测成骨相关基因的表达,观察其对MC3T3-E1细胞成骨向分化的影响;构建大鼠颅骨骨缺损模型,4周和8周后观察GelMA和GelMA-DMOG水凝胶对骨缺损的修复效果。结果:GelMA-DMOG水凝胶孔径大小、分布及降解时间与GelMA水凝胶相比没有明显差异;两种水凝胶对MC3T3-E1细胞增殖均无明显影响;与培养在24孔板上的对照组相比,GelMA组可促进MC3T3-E1细胞成骨向分化,而GelMA-DMOG组明显增强了细胞的成骨向分化和成骨相关基因Runx2、ALP和骨钙素的表达;体内实验发现与GelMA组相比,GelMA-DMOG水凝胶处理组具有明显的促成骨成血管作用和较好的骨缺损修复效果。结论:GelMADMOG水凝胶相对于GelMA材料自身的理化性能没有改变,且体外和体内实验显示该材料对于骨缺损修复具有较好的促进作用,为骨组织工程治疗骨缺损的临床应用开辟了新途径。
Objective:To investigate the effect of dimethyloxallyl glycine(DMOG)-containing gelatin methacryloyl(GelMA) hydrogel on promoting osteogenic differentiation in vitro,and the effect of repairing calvarial bone defects in rats in vivo.Methods:After synthesizing GelMA-DMOG hydrogel,the surface characteristics were observed by scanning electron microscope,and its degradation time was detected in vitro.Mouse osteoblast precursor cells MC3T3-E1 were cultured on the surface of GelMA and GelMADMOG hydrogel.The cytoskeleton morphology was observed by fluorescence,and its cytotoxicity was detected by CCK-8 assay.After cultured cells in osteogenesis induction medium,alkaline phosphatase(ALP),alizarin red staining and qRT-PCR were used to detect the effect of two kinds of hydrogel on osteogenic differentiation of MC3T3-E1 cells.A rat calvarial bone defect model was constructed.The effects of GelMA and GelMA-DMOG hydrogels on bone defect repair were observed after 4 and 8 weeks.Results:Compared with GelMA hydrogel,there was no significant difference in the pore size,distribution,and degradation time of GelMA-DMOG hydrogel.The two groups of hydrogel had no significant effect on the proliferation of MC3T3-E1 cells.Compared with control group,the GelMA hydrogel cultured group slightly promoted osteogenic differentiation of MC3T3-E1 cells,while the GelMA-DMOG hydrogel cultured group significantly enhanced the osteogenic differentiation and the expression of osteogenesis-related genes,such as Runx2,ALP,and osteocalcin(OCN).In vivo,compared with control and GelMA group,the GelMA-DMOG hydrogel group has a much better calvarial bone repair effect.Conclusion:The physical and chemical properties of GelMA-DMOG hydrogel are not changed compared to GelMA hydrogel.In vitro and in vivo experiments show that the GelMA-DMOG hydrogel can promote the repair of bone defects,which opens up a new clinical application of bone tissue engineering in the treatment of bone defects.
作者
张燕萍
陈花花
顾永春
周培刚
ZHANG Yanping;CHEN Huahua;GU Yongchun;ZHOU Peigang(Department of Stomatology,Suzhou Ninth People’s Hospital,Suzhou 215200,China)
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2020年第10期1449-1455,共7页
Journal of Nanjing Medical University(Natural Sciences)
基金
苏州市第九人民医院院级科研基金(院201613)
作者简介
通信作者:张燕萍,E⁃mail:yanpingzhangsz@163.com
