Perfusion-decellularization was an interesting technique to generate a natural extracellular matrix (ECM) with the complete three-dimensional anatomical structure and vascular system. In this study, the esophageal ECM (E-ECM) scaffold was successfully constructed by perfusion-decellularized technique through the vascular system for the first time. And the physicochemical and biological properties of the E-ECM scaffolds were evaluated. The bone marrow mesenchymal stem cells (BMSCs) were induced to differentiate into myocytes in vitro. E-ECM scaffolds reseeded with myocytes were implanted into the greater omenta to obtain recellular esophageal ECM (RE-ECM), a tissue-engineered esophagus. The results showed that the cells of the esophagi were completely and uniformly removed after perfusion. E-ECM scaffolds retained the original four-layer organizational structure and vascular system with excellent biocompatibility. And the E-ECM scaffolds had no significant difference in mechanical properties comparing with fresh esophagi, p > 0.05. Immunocytochemistry showed positive expression of α-sarcomeric actin, suggesting that BMSCs had successfully differentiated into myocytes. Most importantly, we found that in the RE-ECM muscularis, the myocytes regenerated linearly and continuously and migrated to the deep, and the tissue vascularization was obvious. The cell survival rates at 1 week and 2 weeks were 98.5 3.0% and 96.4 4.6%, respectively. It was demonstrated that myocytes maintained the ability for proliferation and differentiation for at least 2 weeks, and the cell activity was satisfactory in the RE-ECM. It follows that the tissue-engineered esophagus based on perfusion-decellularized technique and mesenchymal stem cells has great potential in esophageal repair. It is proposed as a promising alternative for reconstruction of esophageal defects in the future.

灌注脱细胞是一种有趣的技术，可生成具有完整三维解剖结构和血管系统的天然细胞外基质 (ECM)。本研究首次通过血管系统灌注脱细胞技术成功构建了食管ECM（E-ECM）支架。并对 E-ECM 支架的理化和生物学特性进行了评估。体外诱导骨髓间充质干细胞（BMSCs）分化为肌细胞. 将重新植入肌细胞的 E-ECM 支架植入大网膜以获得再细胞食管 ECM (RE-ECM)，这是一种组织工程食管。结果表明，灌注后食管细胞被完全、均匀地去除。E-ECM支架保留了原有的四层组织结构和血管系统，具有优异的生物相容性。与新鲜食管相比，E-ECM支架的力学性能无显着差异，p > 0.05。免疫细胞化学显示α阳性表达-sarcomeric 肌动蛋白，表明 BMSCs 已成功分化为肌细胞。最重要的是，我们发现在RE-ECM肌层中，肌细胞线性连续再生并迁移至深层，组织血管化明显。1周和2周的细胞存活率分别为98.5±3.0%和96.4±4.6%。结果表明，肌细胞至少保持了2周的增殖和分化能力，并且在RE-ECM中细胞活性令人满意。由此可见，基于灌注脱细胞技术和间充质干细胞的组织工程食管在食管修复中具有巨大的潜力。它被提议作为未来食管缺损重建的有希望的替代方案。