Tissue-engineered esophagus: recellular esophageal extracellular matrix based on perfusion-decellularized technique and mesenchymal stem cells

Abstract

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.