【摘要】 背景:许多研究发现三维结构的纳米材料能促进细胞增殖。目的:构建一种新型纳米金/壳聚糖仿生功能支架,观察角质形成细胞在支架上的贴壁、增殖情况。设计、时间及地点:细胞观察性实验,于2006-05/2007-04在南通大学公共卫生学院进行。材料:1.0～2.0d龄SD大鼠由南通大学实验动物中心提供,壳聚糖(脱乙酰度>90%)由浙江省玉环县化工厂提供,氯金酸由上海试剂一厂提供。方法:用化学还原法制备纳米金胶,通过自组装技术将纳米金胶组装到壳聚糖膜表面,得到纳米金/壳聚糖膜仿生功能支架。将SD大鼠皮肤用2.5g/L胰酶消化得到角质形成细胞。将角质形成细胞分别接种在纳米金/壳聚糖仿生功能支架、壳聚糖膜、空白培养板上,计算培养6,12,24h后的贴壁率,用免疫组织化学和透射电子显微镜等手段对培养的角质形成细胞进行鉴定。主要观察指标:角质形成细胞在纳米金/壳聚糖膜上的的贴壁率;纳米金/壳聚糖膜上的角质形成细胞形态。结果:①6,12,24h角质形成细胞在纳米金/壳聚糖仿生功能支架上的贴壁率分别为25.52%,40.34%,58.72%,12,24h贴壁率明显高于其他两组。②免疫组织化学检测结果显示培养的细胞绝大部分为角质形成细胞。③透射电子显微镜观察可以看到角质形成细胞呈圆形,核大,异染色质较多,胞浆内富含线粒体,细胞活力强。结论:纳米金/壳聚糖仿生功能支架能显著提高角质形成细胞的贴壁、生长。更多还原

【Abstract】 BACKGROUND:Studies have reported that nanometer materials at a three-dimensional structure may promote cell proliferation.OBJECTIVE:To construct nano-sized gold colloid/chitosan scaffold and to observe the adhesion and proliferation of keratinocytes.DESIGN,TIME AND SETTING:An observation study was performed in the School of Public Health,Nantong Univerisity from May 2006 to April 2007.MATERIALS:SD rats of 1-2 days old were offered by the Experimental Animal Center of Nantong University,chitosan(deacetylation degree>90%)was purchased from Yuhuan Chemical Engineering Company of Zhejiang Province,and chlorauric acid was produced by the Shanghai No.1 Reagent Factory.METHODS:Nano-sized gold colloid was prepared by chemical reduction method.Nano-sized gold colloid/chitosan was fabricated by self-assembly technology.The keratinocytes were obtained from SD rats using 2.5 g/L trypsin digestion,and then cultured on three different extracellular matrices:nano-sized gold colloid/chitosan scaffold,chitosan film and cell culture plastic.At 6,12,24 hours after inoculation,the cell attached ratios were calculated.The cultured keratinocytes were identified and characterized by immunohistochemistry and transmission electron microscope.MAIN OUTCOME MEASURES:The attached ratio of keratinocyte cultured on nano-sized gold colloid/chitosan scaffold;cell morphology of the cultured keratinocytes on nano-sized gold colloid/chitosan scaffold.RESULTS:The attached ratios after inoculation 6,12,24 hours on the scaffold were 25.52%,40.34%,58.72%,respectively.And the attached ratios of 12 and 24 hours on the scaffold were significantly higher than that on the other two matrices.The result of immunohistochemistry demonstrated that the cells on the scaffold were mainly keratinocytes.It was showed that the keratinocytes were round and had big nuclei,many heterochromatin,mitochondrion in plasma and maintained strong cell activity well through transmission electron microscope.CONCLUSION:Nano-sized gold colloid/chitosan scaffold can promote attachment and proliferation of keratinocytes.更多还原