灌注法制备肺脱细胞基质

Pulmonary perfusion for preparation of the acellular matrix

背景:对组织器官进行脱细胞,细胞外基质在经过去除细胞和可溶性蛋白处理之后,可维持正常的器官外形及组织结构基质成分。目的:利用灌注法制备肺脱细胞基质。方法:将40只Wistar大鼠随机均分为2组,常规麻醉处理后打开胸腔,获得完整的肺组织,实验组利用Langendorff灌注模型构建大鼠全肺脱细胞基质支架,对照组不予以特殊处理。动态观察和记录实验组肺脏颜色和形态变化,分别从2组大鼠肺脏不同部位获取小块组织,利用电子显微镜进行组织学观察,观察两组的Weigert弹力纤维联合Von Gieson结缔组织染色和苏木精-伊红染色情况。结果与结论:经1%脱氧胆酸钠溶液灌注后,实验组大鼠肺脏颜色逐渐发生改变,表现为从内至外呈分段分叶状逐渐变为白色半透明状,并可观察到清晰肺叶结构,最终肺脏呈现出均一的白色半透明状。经Weigert弹力纤维联合Von Gieson结缔组织染色和苏木精-伊红染色发现,对照组新鲜肺组织切片中含有大量细胞,其中包括毛细血管和成纤维细胞及内皮细胞等,细胞呈整齐排列状,具有完整的肺泡结构,弹性纤维结构清晰,胶原纤维也呈整齐排列状,结构较为紧凑;实验组肺组织细胞基本已消失,仍具有完整的肺泡形态结构,但呈较为疏松的状态且裂隙增大,弹性纤维保存良好,胶原纤维呈较疏松排列状。表明利用灌注法课可有效构建大鼠全肺组织基质支架。

BACKGROUND：The extracelular matrix with removal of cels and soluble proteins can maintain the normal shape of organs and matrix components. OBJECTIVE：To prepare the acelular matrix of lung tissue using perfusion method. METHODS：Forty Wistar rats were randomly divided into two groups： under routine anesthesia, the chest was open to obtain complete lung tissue for construction of rat lung acelular matrix scaffold using Langendorff perfusion model in experimental group, and there was no treatment in control group. Lung tissue color and shape were observed and recorded dynamicaly. A smal tissue from different sites of the lung was taken from each group and observed histologicaly under electron microscope. The elastic fibers and connective tissues were highlighted by a Weigert or von Gieson staining, respectively. Hematoxylin-eosin staining was also employed. RESULTS AND CONCLUSION：After perfusion with 1% sodium deoxycholate, the lung tissue of the experimental group gradualy appeared with a piecewise and lobulated translucent appearance in white color from the inside to the outside, and the lung structure was clear. Eventualy, the lung tissue became uniform white translucent. The Weigert-Von Gieson staining and hematoxylin-eosin staining showed that there were a large amount of cels in fresh lung tissue sections of the control group, including capilaries, fibroblasts and endothelial cels; the cels were arranged neatly, and had complete alveolar structure; the elastic fiber structure was clear, and the colagen fiber was arranged neatly and compactly. In the experimental group, the lung cels nearly disappeared, and the alveolar structure was stil intact but in a loose state; the elastic fibers were preserved wel and the colagen fibers were loosely arranged. These findings indicate that the perfusion method can be used to effectively construct the lung matrix scaffold in rats.