【摘要】 设计合成了3种不同序列的基质金属蛋白酶(MMP)敏感肽,与活性基团马来酰亚胺修饰后的透明质酸进行共价交联,制备一类对MMP具有响应性的透明质酸仿生杂化水凝胶,并研究其对诱导兔骨髓间充质干细胞(BMSCs)向软骨方向分化的影响。以N-羟基硫代琥珀酰亚胺(Sulfo-NHS)与1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC·HCl)为催化剂,活化透明质酸羧基后,与N-(2-氨基乙基)马来酰亚胺三氟乙酸盐(AEM)常温搅拌反应,得到接枝马来酰亚胺基团的透明质酸(HA-MAL),马来酰亚胺基团与MMP敏感肽上的巯基在常温下即可发生加成反应。共价交联MMP敏感肽的水凝胶具有良好的机械性能,能够包埋细胞3D培养达4周以上。对水凝胶3D及孔板2D培养的BMSCs进行实时荧光定量PCR测定软骨方向分化的特征性基因Marker表达,结果显示,3种水凝胶对COL1A2、COL2A1、ACAN、SOX9 4种基因表达具有不同程度的上调作用,可促进BMSCs的软骨分化。更多还原

【Abstract】 Three different sequences of matrix metalloproteinases(MMP)sensitive peptides were designed and covalently crosslinked with hyaluronic acid which was modified with active group of maleimide to prepare a class of hyaluronic acid bionic hybrid hydrogel and then its effect on the cartilage differentiation of rabbit bone marrow mesenchymal stem cells(BMSCs)was studied.The hyaluronic acid(HA)was activated with(Sulfo-NHS)and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(EDC·HCl)as catalyst,then N-(2-aminoethyl)maleimide trifluoroacetate(AEM)was grafted with activated HA at room temperature to obtain HAMAL.The addition reaction of the maleimide group with the mercapto group on the MMP-sensitive peptide could be carried out at room temperature.Covalently cross-linked MMP-sensitive peptide hydrogel has good mechanical properties,cells can be embedded in which for more than 4 weeks.The results show that compared with 2 Dorifice plate cells culture,hydrogels have different effects on the up regulation of COL1 A2,COL2 A1,ACAN and SOX9 gene expression,which can promote BMSCs to cartilage differentiation.更多还原