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组织工程食管:基于灌流去细胞技术和间充质干细胞的再细胞化食管细胞外基质。

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

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan Province, People's Republic of China.

Department of Otorhinolaryngology Head and Neck Surgery, First Affiliated Hospital, Chengdu Medical College, Chengdu, Sichuan Province, People's Republic of China.

出版信息

Biomed Mater. 2021 Aug 27;16(5). doi: 10.1088/1748-605X/ac1d3d.

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. 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,> 0.05. Immunocytochemistry showed positive expression of-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.

摘要

灌流去细胞技术是一种有趣的技术,可以生成具有完整三维解剖结构和脉管系统的天然细胞外基质(ECM)。在这项研究中,首次通过脉管系统成功构建了灌流去细胞技术的食管 ECM(E-ECM)支架。并评估了 E-ECM 支架的理化和生物学特性。骨髓间充质干细胞(BMSCs)被诱导分化为肌细胞。将 E-ECM 支架与肌细胞重新接种到更大的网膜中,获得了具有细胞的食管 ECM(RE-ECM),即组织工程食管。结果表明,灌流后,食管细胞被完全均匀地去除。E-ECM 支架保留了原始的四层组织结构和脉管系统,具有优异的生物相容性。并且 E-ECM 支架的机械性能与新鲜食管无显著差异,>0.05。免疫细胞化学显示-sarcomeric 肌动蛋白呈阳性表达,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,肌细胞呈线性和连续再生并向深部迁移,组织血管化明显。第 1 周和第 2 周的细胞存活率分别为 98.5±3.0%和 96.4±4.6%。这表明肌细胞至少在 2 周内保持增殖和分化的能力,RE-ECM 中的细胞活性令人满意。因此,基于灌流去细胞技术和间充质干细胞的组织工程食管在食管修复方面具有巨大潜力。它被提出作为未来食管缺损重建的一种有前途的替代方法。

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