大鼠胰腺去细胞天然支架生物相容性的鉴定

发布时间：2018-05-18 01:05

本文选题：去细胞 + 胰腺　；参考：《解剖学报》2014年04期

【摘要】：目的通过离体灌注化学去垢剂的方法制备大鼠胰腺去细胞天然生物支架,并对支架的完整性、生物相容性进行检验。方法健康成年SD大鼠30只,分别经胆管与血管两条途径灌注十二烷基硫酸钠和曲亚通X-100等药品洗脱,获取胰腺去细胞生物支架。通过HE染色、扫描电子显微镜和透射电子显微镜等观察细胞残留于去细胞支架的生物学特性,分光光度计法鉴定残留去垢剂量,酶联免疫吸附剂测定法测定残留支架蛋白含量及生长因子,并用腹壁与皮下包埋实验检验其炎症反应,MTT法测定支架细胞毒性,内皮细胞共培养测定支架生物相容性。结果 HE染色、扫描电子显微镜和透射电子显微镜观察均表明去细胞支架无细胞残留,细胞外支架连续性完好,脉管支架保存完整;药物残留小于规定标准;细胞外支架生长因子及支架蛋白存留量等指标上血管组均优于胆管组;腹壁与皮下包埋实验显示,去细胞支架炎性反应较对照组明显减轻;MTT法显示无细胞毒性;内皮细胞共培养有黏附趋势。结论使用两种灌注法制备胰腺去细胞生物支架,均可将细胞去除彻底,支架生物相容性好。但就血管组细胞外支架保留完整性,生长因子保留更多,则血管灌注途径优于胆管灌注途径。
[Abstract]:Aim to prepare the acellular natural scaffold of rat pancreas by in vitro infusion of chemical defouling agent, and to test the integrity and biocompatibility of the scaffold. Methods 30 healthy adult SD rats were perfused with drugs such as sodium dodecyl sulfate and triazone X-100 respectively through bile duct and blood vessel to obtain acellular pancreatic scaffold. He staining, scanning electron microscope and transmission electron microscope were used to observe the biological characteristics of cells remaining in the scaffold, and the residual defouling dose was identified by spectrophotometer. The contents of residual stents protein and growth factors were determined by Elisa. The cytotoxicity of stents was determined by MTT assay and the biocompatibility of stents was determined by co-culture of endothelial cells. Results HE staining, scanning electron microscopy and transmission electron microscopy showed that there were no cell residues in the acellular stents, the extracellular stents were intact, the vascular stents were intact, and the drug residues were less than the prescribed standard. The upper vascular group was superior to the bile duct group in the indexes of extracellular stent growth factor and stenting protein retention, and the inflammatory response of acellular stents in abdominal wall and subcutaneous embedding was significantly less than that in the control group, and no cytotoxicity was observed by MTT assay. The co-culture of endothelial cells has the tendency of adhesion. Conclusion the acellular scaffolds of pancreas prepared by two kinds of perfusion methods can remove the cells completely and have good biocompatibility. However, the vascular perfusion pathway is superior to the bile duct perfusion pathway for the integrity of extracellular stent and more growth factors in vascular group.
【作者单位】：温州医科大学第一临床医学院;温州医科大学生物支架移植与免疫研究所;温州医科大学基础医学中心;
【基金】：国家级大学生创新创业训练计划资助项目(201310343006,201210343008) 浙江省自然科学基金资助项目(LY12H15002) 浙江省大学生科技创新活动计划“新苗人才计划”资助项目(2012R413004) 浙江省教育厅资助项目(Y201327980)
【分类号】：R318.08

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