可注射葡萄糖酸内酯-海藻酸钠/β-磷酸三钙-聚乙二醇复合水凝胶支架的性能评价  

Properties of injectable gluconolactone-sodium alginate/beta-tricalcium phosphate/polyethylene glycol composite hydrogel scaffold

作　　者：刘印 刘琴[1,2,3] 陈娇 古先扬 陈佳文 马敏先 Liu Yin;Liu Qin;Chen Jiao;Gu Xianyang;Chen Jiawen;Ma Minxian(Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Stomatological Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Research Center of Tissue Engineering and Stem Cell Technique,Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Department of Oral Implantology,Guiyang Hospital of Stomatology,Guiyang 550004,Guizhou Province,China;National-Guizhou Joint Engineering Laboratory of Cell Engineering and Biomedicine Technique,Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学,贵州省贵阳市550004 [2]贵州医科大学附属口腔医院,贵州省贵阳市550004 [3]贵州医科大学组织工程与干细胞实验中心,贵州省贵阳市550004 [4]贵阳市口腔医院口腔种植科,贵州省贵阳市550004 [5]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,贵州省贵阳市550004

出　　处：《中国组织工程研究》2022年第27期4308-4313,共6页Chinese Journal of Tissue Engineering Research

基　　金：贵阳市科技计划项目(筑科合同[2018]1-81号),项目负责人:马敏先。

摘　　要：背景:可注射海藻酸钠水凝胶可以通过非侵入性或微创方式修复骨缺损,但力学性能欠佳,聚乙二醇基水凝胶具有弹性,将材料复合有望提高水凝胶支架的机械性能及细胞生物相容性。目的:探索葡萄糖酸内酯-海藻酸钠/β-磷酸三钙-聚乙二醇复合水凝胶的理化性能,以及对鼠骨髓间充质干细胞增殖和分化的影响。方法:以葡萄糖酸内酯为交联剂,分别制备葡萄糖酸内酯-海藻酸钠/β-磷酸三钙-聚乙二醇水凝胶(交联剂的浓度分别为5,10,20 g/L,依次记为A、B、C组)与葡萄糖酸内酯-海藻酸钠/β-磷酸三钙水凝胶(交联剂的浓度为10 g/L,记为D组),表征4组水凝胶的形态、机械性能与凝胶时间。将SD大鼠骨髓间充质干细胞分别与4组水凝胶共培养,利用CCK-8法检测细胞增殖,Live/Dead荧光染色观察细胞存活情况,Ⅰ型胶原免疫细胞化学染色检测成骨分化情况。结果与结论:①扫描电镜下,A、B、C组可见类似黏结性的丝状结构,A组孔径分布不均匀,B组孔径分布均匀且孔隙率高,C组孔隙率高但孔径大小不一,D组孔径分布不均且孔隙率低。②理化性质:B组的压缩应力高于D组(P<0.05),并且随着交联剂浓度的增加,A、B、C组支架的压缩应力增强(P<0.05);压缩50%后水凝胶的大体观显示,压缩后D组有较大裂纹,B组仅有较小裂纹;A、B、C组凝胶时间随加入交联剂浓度的增加而缩短,B组凝胶时间较D组稍延长(P<0.05)。③CCK-8实验检测显示,A、B组的细胞增殖速率高于C组(P<0.05),B组高于D组(P<0.05);Live/Dead染色显示,各组细胞存活率均较高,但C组死细胞较其他3组多。④免疫细胞化学染色显示,A、B、D组细胞呈多角形,C组细胞部分呈圆形,B组绿色荧光强度较其他3组高,且细胞骨架和微丝的形态及细胞数量也优于其他3组。⑤结果表明,葡萄糖酸内酯-海藻酸钠/β-磷酸三钙-聚乙二醇水凝胶具有良好的机械性能和稳定性,其中以10BACKGROUND:Injectable sodium alginate hydrogel can repair bone defects by non-invasive or minimally invasive methods,but the mechanical properties are poor and polyethylene glycol-based hydrogel has elasticity.The composite of materials is expected to improve the mechanical properties and cellular biocompatibility of hydrogel scaffolds.OBJECTIVE:To investigate the physical and chemical properties of glucolactone-sodium alginate/β-tricalcium phosphate-polyethylene glycol hydrogel and its effect on the proliferation and differentiation of murine bone marrow mesenchymal stem cells.METHODS:Gluconolactone as cross-linking agent was set to prepare glucolactone-sodium alginate/β-tricalcium phosphate-polyethylene glycol hydrogel(concentrations of cross-linking agent were 5,10,and 20 g/L,respectively,which were marked as groups A,B,and C)and glucolactone-sodium alginate/β-tricalcium phosphate hydrogel(concentration of cross-linking agent was 10 g/L,which was marked as group D).The morphology,mechanical properties,and gelation time of hydrogels in the four groups were characterized.Rat bone marrow mesenchymal stem cells were co-cultured with four groups of hydrogels.The proliferation of cells was detected by CCK-8 assay.Cell survival was observed by Live/Dead fluorescence staining.Osteogenic differentiation was detected by immunocytochemical staining of type Ⅰ collagen.RESULTS AND CONCLUSION:(1)The filamentous structure similar to adhesion was observed by scanning electron microscope in the groups A,B and C.The pore size distribution of group A was uneven.The pore size of group B was uniform and the porosity was high.The group C had high porosity but different pore sizes.The group D had uneven pore size distribution and low porosity.(2)Physicochemical properties:The compressive stress in group B was higher than that in group D(P<0.05).With the increase of gluconolactone concentration,the compressive stress of hydrogel in groups A,B and C increased(P<0.05).The macroscopic performance of hydrogel after 50%compression sh

关 键 词：组织工程支架 可注射水凝胶 海藻酸钠 Β-磷酸三钙 聚乙二醇 葡萄糖酸内酯 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]