壳聚糖温敏凝胶负载釉基质蛋白对骨髓基质细胞的作用  

Effects of enamel matrix proteins loaded in chitosan thermosensitive hydrogel on bone marrow stromal cells in vitro

作　　者：吴广升[1] 张艺文[2] 王新文[1] 马志伟[1] 曹敏[1] 王勤涛[1] 

机构地区：[1]第四军医大学口腔医学院牙周黏膜病科,陕西西安710032 [2]第四军医大学口腔医学院牙体牙髓病科,陕西西安710032

出　　处：《上海口腔医学》2009年第2期178-182,共5页Shanghai Journal of Stomatology

基　　金：陕西省科学发展计划基金(2006K14-G3)~~

摘　　要：目的:探讨在壳聚糖温敏凝胶支架材料中釉基质蛋白(enamel matrix proteins,EMPs)对骨髓基质细胞(bone marrow stromal cells,BMSCs)增殖和碱性磷酸酶(alkaline phosphate,ALP)活性的影响。方法:合成壳聚糖温敏凝胶并加入适当浓度的EMPs,通过考马斯亮蓝试剂盒检测其对EMPs的缓释作用。用全骨髓培养法获得大鼠BMSCs,含10%胎牛血清的DMEM培养液进行原代培养。含EMPs浓度分别为0、50、100、150μg/mLDMEM培养液培养第3代大鼠BMSCs,MTT法测定各组细胞的增殖活性。MTT法及ALP试剂盒检测大鼠BMSCs在负载100μg/mLEMPs及不负载EMPs的壳聚糖温敏凝胶支架材料中的增殖情况及ALP的活性。采用SPSS11.0软件包对实验数据进行单因素方差分析及两样本t检验。结果:EMPs可在壳聚糖温敏凝胶中持续释放3周以上。DMEM培养液中,50μg/mLEMPs组从实验的第3天开始,显著促进大鼠BMSCs的增殖(P<0.01)。壳聚糖温敏凝胶负载100μg/mLEMPs后,于实验的第3天和第5天明显促进大鼠BMSCs的增殖(P<0.05)并且在实验的第7天(P<0.05)和第9天(P<0.01)显著促进大鼠BMSCsALP水平的提高。结论:壳聚糖温敏凝胶对EMPs具有缓释性,负载EMPs后,可以促进大鼠BMSCs增殖,提高ALP的活性。PURPOSE： To investigate the effects of enamel matrix proteins （EMPs） on proliferation, alkaline phosphate （ALP） activity of bone marrow stromal cells （BMSCs）seeded on the scaffold of chitosan thermosensitive hydrogel. METHODS： Chitosan thermosensitive hydrogel was prepared and its slow-releasing effect of EMPs was checked by eoomassie blue staining kit. Rat BMSCs were obtained from rat bone marrow aspiration and cultured in DMEM medium with 10% fetal bovine serum （FBS）. Rat BMSCs were exposed to various concentrations of EMPs （0,50,100 and 150μg/mL） and their proliferation rates were assessed by MTT assay. The proliferation rates and ALP activity of rat BMSCs were examined by MTT assay and ALP kit when BMSCs cultured on the scaffolds of chitosan thermosensitive hydrogel loading with or without 100μg/mL EMPs .The data was statistically analyzed with SPSS11.0 software package for a parametric one- way analysis of variance （ANOVA） and two-sample t test. RESULTS： The release of EMPs in chitosan thermosensitive hydrogel lasted for more than 3 weeks. In DMEM medium, 50μg/mL EMPs significantly enhanced BMSCs proliferation from day 3 over the experiment （P〈0.01）. In ehitosan thermosensitive hydrogel scaffolds loading 100μg/mL EMPs,both the proliferation at day 3 and 5 （P〈0.05）and the ALP activity at day 7 （P〈0.05）and 9（P〈0.01） of BMSCs in the experiment were promoted. CONCLUSION： EMPs loaded on the chitosan thermosensitive hydrogel exhibits significant effects on proliferation and ALP activity of rat BMSCs. Supported by Science Development Plan of Shanxi Province（Grant No. 2006 K14-G3）.

关 键 词：壳聚糖 温敏凝胶 釉基质蛋白 骨髓基质细胞 

分 类 号：R318.08[医药卫生—生物医学工程]