Laboratory of Tissue Engineering, Rigshospitalet, Faculty of Health and Medical Sciences, University of Copenhagen, Henrik Harpestrengs Vej 4C, 2100, Copenhagen, Denmark.
Division of Pediatric Surgery, Department of Surgery and Transplantation, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
Sci Rep. 2023 Aug 21;13(1):13589. doi: 10.1038/s41598-023-40049-0.
Autologous micrografting is a technique currently applied within skin wound healing, however, the potential use for surgical correction of other organs with epithelial lining, including the urinary bladder, remains largely unexplored. Currently, little is known about the micrograft expansion potential and the micromolecular events that occur in micrografted urothelial cells. In this study, we aimed to evaluate the proliferative potential of different porcine urothelial micrograft sizes in vitro, and, furthermore, to explore how urothelial micrografts communicate and which microcellular events are triggered. We demonstrated that increased tissue fragmentation subsequently potentiated the yield of proliferative cells and the cellular expansion potential, which confirms, that the micrografting principles of skin epithelium also apply to uroepithelium. Furthermore, we targeted the expression of the extracellular signal-regulated kinase (ERK) pathway and demonstrated that ERK activation occurred predominately at the micrograft borders and that ERK inhibition led to decreased urothelial migration and proliferation. Finally, we successfully isolated extracellular vesicles from the micrograft culture medium and evaluated their contents and relevance within various enriched biological processes. Our findings substantiate the potential of applying urothelial micrografting in future tissue-engineering models for reconstructive urological surgery, and, furthermore, highlights certain mechanisms as potential targets for future wound healing treatments.
自体微移植是一种目前应用于皮肤创伤愈合的技术,然而,其在外科矫正其他具有上皮衬里的器官(包括膀胱)方面的潜在用途在很大程度上仍未得到探索。目前,人们对微移植上皮细胞的微移植扩展潜力和微观分子事件知之甚少。在这项研究中,我们旨在评估不同猪膀胱微移植大小的体外增殖潜力,并且进一步探索微移植上皮细胞如何相互交流以及触发哪些微观细胞事件。我们证明,组织碎片的增加随后增强了增殖细胞的产量和细胞的扩增潜力,这证实了皮肤上皮的微移植原则也适用于尿路上皮。此外,我们针对细胞外信号调节激酶(ERK)通路的表达进行了研究,并证明 ERK 的激活主要发生在微移植的边缘,而 ERK 的抑制导致尿路上皮迁移和增殖减少。最后,我们从微移植培养基中成功分离出外泌体,并评估了它们在各种富含生物学过程中的含量和相关性。我们的研究结果为将来在重建泌尿外科手术的组织工程模型中应用尿路上皮微移植提供了依据,并进一步强调了某些机制作为未来伤口愈合治疗的潜在靶点。