可注射壳聚糖温敏水凝胶对犬骨髓基质细胞生物学行为的影响  

Effects of injectable chitosan thermosensitive hydrogel on dog bone marrow stromal cells in vitro

作　　者：韩雪[1,2] 刘洪臣[1] 王东胜[1] 苏方[1] 鄂玲玲[1] 吴霞[1] 

机构地区：[1]解放军总医院口腔医学研究所,北京100853 [2]沈阳军区总医院口腔科,辽宁沈阳110016

出　　处：《上海口腔医学》2011年第2期113-118,共6页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(30973354)~~

摘　　要：目的:探讨可注射壳聚糖(chitosan,CS)基温敏水凝胶对犬骨髓基质细胞(bone marrow stromal cells,BMSCs)生物学活性的影响。方法:利用密度梯度离心法获得犬BMSCs,含10%胎牛血清的L-DMEM培养液进行原代培养并进行初步鉴定。合成壳聚糖温敏水凝胶,光镜下观察BMSCs在凝胶中的存活情况。制备凝胶浸提液,检测其对细胞增殖分化的影响。采用SPSS13.0软件包对实验数据进行单因素方差分析。结果:犬BMSCs可在壳聚糖温敏凝胶中存活。壳聚糖浸提液组对BMSCs的增殖与正常培养液组相比,无显著性差异;但在培养第1、4、7天时,矿化培养液组和矿化浸提液组的细胞增殖率明显低于未矿化的完全培养液组和壳聚糖浸提液组,第10天时,4组间无显著性差异。壳聚糖矿化浸提液组碱性磷酸酶和骨钙素活性明显提高。结论:制备的壳聚糖温敏水凝胶具有良好的生物相容性,可进行水凝胶复合BMSCs的体内外实验研究。PURPOSE： To investigate the biological effects of injectable chitosan（CS） thermosensitive hydrogel on dog bone marrow stromal cells（BMSCs） in vitro.METHODS： Dog BMSCs were obtained from dog bone marrow aspiration by density gradient centrifugation method,cultured in DMEM medium with 10% fetal bovine serum（FBS） and identified.Chitosan thermosensitive hydrogel was prepared and the BMSCs cultured in a medium containing hydrogel were observed under light microscope.The extract liquid from hydrogel was made and its effects on cell proliferation and differentiation were evaluated.Statistical significance was assessed with SPSS 13.0 software package for one-way ANOVA.RESULTS： Dog BMSCs survived in the extract liquid and there was no significant difference.The proliferation of BMSCs in mineralization condition medium was lower than normal medium after 1,4,7 days culture and was similar to that in normal medium after 10 days.The osteogenic extract liquid improved the secretion of alkaline phosphatase（ALP） and osteocalcin（OC）.CONCLUSIONS： The CS thermosensitive hydrogel possesses good biocompatibility and can be used for in vitro and in vivo experimental studies of CS hydrogel loaded with BMSCs.

关 键 词：壳聚糖 温敏水凝胶 细胞培养 骨髓基质细胞 

分 类 号：R318.08[医药卫生—生物医学工程]