Centre de recherche du CHU de Québec - Université Laval, Department of Surgery, Faculty of Medicine, Université Laval and Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Canada.
Department of Molecular Biology, Medical Biochemistry and Pathology, Faculty of Medicine, Université Laval, Canada.
Acta Biomater. 2019 May;90:192-204. doi: 10.1016/j.actbio.2019.04.010. Epub 2019 Apr 4.
Organs are needed for the long-term replacement of diseased or wounded tissues. Various technologies based on cells seeded in synthetic or biomaterial scaffolds, or scaffold-free methods have been developed in order to produce substitutes that mimic native organs and tissues. For cell-based approaches, the use of living allogeneic fibroblasts could potentially lead to the production of "off-the-shelf" bioengineered organs/tissues. However, questions remain regarding the outcome of allogeneic grafts in terms of persistence of allogeneic cells, tolerance and the host immune reaction against the tissue after implantation. To evaluate graft tolerance of engineered-tissues containing non-autologous fibroblasts, tissue-engineered skin substitutes (TESs) produced with syngeneic, allogeneic or xenogeneic fibroblasts associated with syngeneic, allogeneic or xenogeneic epithelial cells were grafted in mice as primary and secondary grafts. The immune response was evaluated by histological analysis and immunodetection of M2 macrophages, CD4- and CD8-positive T cells, 15, 19, 35 and 56 days after grafting. Tissue-engineered skin composed of non-autologous epithelial cells were rejected. In contrast, TESs composed of non-autologous fibroblasts underlying syngeneic epithelial cells were still present 56 days after grafting. This work shows that TES composed of non-autologous fibroblasts and autologous epithelial cells are not rejected after grafting. STATEMENT OF SIGNIFICANCE: We found that tissue-engineered skin substitutes produced by a scaffold-free cell-based approach from allogeneic fibroblasts and autologous epithelial cells are not rejected after grafting and allow for the permanent coverage of a full-thickness skin wounds. In the field of tissue engineering, these findings open the possibility of selecting a human fibroblastic or stromal cell population based on its biological properties and adequate biosafety, banking it, in order to produce "ready-to-use" bioengineered organs/tissues that could be grafted to any patient without eliciting immune reaction after grafting. Our results can be generalized to any organs produced from fibroblasts. Thus, it is a great step with multiple applications in tissue engineering and transplantation.
器官需要长期替代患病或受伤的组织。为了生产出模仿天然器官和组织的替代品，已经开发出了各种基于细胞接种在合成或生物材料支架上的技术，或无支架方法。对于基于细胞的方法，使用活的同种异体成纤维细胞有可能导致“现成的”生物工程器官/组织的生产。然而，关于同种异体移植物的结果，仍然存在一些问题，如同种异体细胞的持久性、耐受性以及宿主对植入后的组织的免疫反应。为了评估含有非自体成纤维细胞的工程组织的移植物耐受性，将用同基因、同种异体或异种成纤维细胞与同基因、同种异体或异种上皮细胞联合制成的组织工程皮肤替代物（TES）作为原发性和继发性移植物在小鼠中进行移植。通过组织学分析和 M2 巨噬细胞、CD4+和 CD8+T 细胞的免疫检测，在移植后 15、19、35 和 56 天评估免疫反应。由非自体上皮细胞组成的组织工程皮肤被排斥。相比之下，由非自体成纤维细胞和同基因上皮细胞组成的 TES 在移植后 56 天仍存在。这项工作表明，由非自体成纤维细胞和自体上皮细胞组成的 TES 在移植后不会被排斥。
我们发现，由同种异体成纤维细胞和自体上皮细胞通过无支架细胞方法制成的组织工程皮肤替代物在移植后不会被排斥，并允许永久性覆盖全层皮肤伤口。在组织工程领域，这些发现为选择基于其生物学特性和适当生物安全性的人类成纤维细胞或基质细胞群体提供了可能性，将其储存起来，以便生产出“即用型”的生物工程器官/组织，在移植后不会引起免疫反应，可以用于任何患者。我们的研究结果可以推广到任何由成纤维细胞产生的器官。因此，这是组织工程和移植领域的一个重大进展，具有多种应用。
