适用于不同尺寸血管的脱细胞方法研究

将胰酶消化与反复冻融相结合，旨在建立一种适用于各种类型血管的通用脱细胞方法，如隐静脉、颈动脉和主动脉。隐静脉、颈动脉和主动脉经胰酶消化和反复冻融脱细胞处理后，采用苏木精—伊红染色、Masson三色染色及弹性纤维染色来定性评价脱细胞效果和细胞外基质的保存效果，采用Image-Pro-Plus 5.1图像处理软件作进一步定量评价；扫描电子显微镜观察胞外基质的完整性。结果显示，组织染色及定量分析表明此胰酶消化与反复冻融相结合的方法完全脱除了隐静脉、颈动脉和主动脉得细胞，细胞外介质结构保存良好且完整。扫描电子显微镜观察亦表明细胞外基质保存良好，且基质纤维致密规整。表明胰酶消化与反复冻融相结合的脱细胞方法是一种很有前景的制备各种不同类型血管支架的方法。

A general decellularization method for preparing vascular scaffolds from blood vessels of different types, diameters and wall-thicknesses

This study was to build a general decellularization method by combining trypsinization with repeated frozen/thawing treatment for different types of vessels including saphenous vein, carotid artery and aorta. Saphenous vein, carotid artery and aorta were decellularized by trypsinization and repeated frozen/thawing treatment. The efficiencies of cell removal and extracellular matrix (ECM) integrity were examined by Hematoxylin and eosin staining, Masson Trichrome staining and Weigert’s staining. A quantitative means based on image analysis software was used to quantify the ECM preservation. Scanning electron microscopy was used to show the extracellular matrix integrity. Histology staining and quantitative analysis demonstrated trypsinization with repeated frozen/thawing treatment decellularized the saphenous vein, carotid artery and aorta completely. The ECM structure was optimally preserved and integrity. Scanning electron microscopy examination also showed the ECM was well-preserved and the fibers were dense and orderly. The present results revealed that the trypsinization with repeated frozen/thawing method is a promising one for preparing decellularized vascular scaffolds of different types, sizes and wall-thicknesses of blood vessels.