目的从鱿鱼软骨中分离纯化非变性II型胶原蛋白,并对其进行结构表征。方法将鱿鱼软骨冻干后粉碎成200目软骨粉,用4%(w/w)Na OH溶液去除多糖、0.5 M EDTA溶液脱灰对软骨进行前处理,采用胃蛋白酶水解提取,Na Cl盐析用纯水透析后冷冻干燥得...目的从鱿鱼软骨中分离纯化非变性II型胶原蛋白,并对其进行结构表征。方法将鱿鱼软骨冻干后粉碎成200目软骨粉,用4%(w/w)Na OH溶液去除多糖、0.5 M EDTA溶液脱灰对软骨进行前处理,采用胃蛋白酶水解提取,Na Cl盐析用纯水透析后冷冻干燥得到II型胶原蛋白样品。通过氨基酸组成分析、SDS-PAGE电泳、紫外吸收光谱、电镜扫描等对制备的样品鉴定。结果制备的II型胶原蛋白具有3条α1链,分子量112 k Da,在224.5 nm波长处有最大紫外吸收,具有三螺旋结构特征。展开更多
In order to evaluate the cytocompatibility and biocompatibility of a new kind of chitosan blend film as a carrier of corneal endothelial cell, rabbit corneal endothelial cells cultured in vitro were breeded onto the f...In order to evaluate the cytocompatibility and biocompatibility of a new kind of chitosan blend film as a carrier of corneal endothelial cell, rabbit corneal endothelial cells cultured in vitro were breeded onto the film. After a cell monolayer formed, the scanning electron micrography was performed. After inplanted into anterior chamber, slit lamp observation, thickness metering, specular microscopy and HE staining were performed at random time after operation to evaluate the biocompatibility. Inflmmnation in anterior, thickness of cornea, cell density, hexagonality and cell size of the surgical cornea were taken as the indexes of biocompatibility. The cultured cells exhibited a confluent monolayer 10 days after incubation, which proved the satisfactory cytocompatibility of this film. Biocompatibility assay results suggested the implantation feasibility of the film as a carder of corneal endothelial cells.展开更多
文摘目的从鱿鱼软骨中分离纯化非变性II型胶原蛋白,并对其进行结构表征。方法将鱿鱼软骨冻干后粉碎成200目软骨粉,用4%(w/w)Na OH溶液去除多糖、0.5 M EDTA溶液脱灰对软骨进行前处理,采用胃蛋白酶水解提取,Na Cl盐析用纯水透析后冷冻干燥得到II型胶原蛋白样品。通过氨基酸组成分析、SDS-PAGE电泳、紫外吸收光谱、电镜扫描等对制备的样品鉴定。结果制备的II型胶原蛋白具有3条α1链,分子量112 k Da,在224.5 nm波长处有最大紫外吸收,具有三螺旋结构特征。
基金the High Technology Research and Development Program of China(2003AA625050)the National Natural Science Foundation of China(30070220)
文摘In order to evaluate the cytocompatibility and biocompatibility of a new kind of chitosan blend film as a carrier of corneal endothelial cell, rabbit corneal endothelial cells cultured in vitro were breeded onto the film. After a cell monolayer formed, the scanning electron micrography was performed. After inplanted into anterior chamber, slit lamp observation, thickness metering, specular microscopy and HE staining were performed at random time after operation to evaluate the biocompatibility. Inflmmnation in anterior, thickness of cornea, cell density, hexagonality and cell size of the surgical cornea were taken as the indexes of biocompatibility. The cultured cells exhibited a confluent monolayer 10 days after incubation, which proved the satisfactory cytocompatibility of this film. Biocompatibility assay results suggested the implantation feasibility of the film as a carder of corneal endothelial cells.
