Human Amniotic Membrane Enriched with Urinary Bladder Fibroblasts Promote the Re-Epithelization of Urothelial Injury

Abstract

Culturing cells in three-dimensional systems that include extracellular matrix components and different cell types mimic the native tissue and as such provide much more representative results than conventional two-dimensional cell cultures. In order to develop biomimetic bladder tissue in vitro, we used human amniotic membrane (AM) extracellular matrix as a scaffold for bladder fibroblasts (BFs) and urothelial cells. Our aims were to evaluate the integration of BFs into the AM stroma, to assess the differentiation of the urothelium on BFs-enriched AM scaffolds, and to evaluate the AM as a urothelial wound dressing. First, to achieve the optimal integration of BFs into AM stroma, different intact and de- epithelialized AM (dAM) scaffolds were tested. BFs secreted matrix metalloproteinase (MMP)-1 and MMP-2 and integrated into the stroma of all types of AM scaffolds. Second, to establish urothelial tissue equivalent, urothelial cells were seeded on dAM scaffolds enriched with BFs. The BFs in the stroma of the AM scaffolds promoted (1) the proliferation of urothelial cells, (2) the attachment of urothelial cells on AM basal lamina with hemidesmosomes, and (3) development of multilayered urothelium with expressed uroplakins and well-developed cell junctions. Third, we established an ex vivo model of the injured bladder to evaluate the dAM as a wound dressing for urothelial full-thickness injury. dAM acted as a promising wound dressing since it enabled rapid re-epithelization of urothelial injury and integrated into the bladder tissue. Herein, the developed urothelial tissue equivalents enable further mechanistic studies of bladder epithelial–mesenchymal interactions, and they could be applied as biomimetic models for preclinical testing of newly developed drugs. Moreover, we could hypothesize that AM may be suitable as a dressing of the wound that occurs during transurethral resection of bladder tumor, since it could diminish the possibility of tumor recurrence, by promoting the rapid re-epithelization of the urothelium.

The Antibacterial Activity of Human Amniotic Membrane against Multidrug-Resistant Bacteria Associated with Urinary Tract Infections: New Insights from Normal and Cancerous Urothelial Models

“…Our previous studies showed that hAM scaffolds enable the development of urothelium with molecular and ultrastructural properties comparable to those of native urothelium [ 92 ] and that hAM scaffolds enriched with the urinary bladder fibroblasts promote the re-epithelization of the urothelial injury [ 117 ]. Therefore, we believe that the hAM homogenate has the potential not only to decrease the number of bacteria, but also to promote the regeneration of the urothelium, allowing for more rapid tissue regeneration after bacterial infection.…”

Understanding of mouse and human bladder at single‐cell resolution: integrated analysis of trajectory and cell‐cell interactive networks based on multiple scRNA‐seq datasets

“…Bladder fibroblasts could promote re‐epithelization after urothelial injury through enhancement for cell proliferation, attachment to the basal lamina, and development of well‐organized cell junction between multilayered urothelial cells. 41 Also, the existence of laminin, collagen, and elastin in the bladder submucosa matrix was maintained as valuable bioactive factors even after the decellularization and extraction processes. 42 For these interstitial cells (eg, fibroblasts), their close proximity to the urothelium and smooth muscle cell (detrusor) seemed to suggest their modulating or bridging role in the bladder wall.…”

Systematic Review of the Application of Perinatal Derivatives in Animal Models on Cutaneous Wound Healing

“…As the most commonly used perinatal tissue is the hAM, we assume that the poor percentage of the in vitro characterization is due to the fact that the hAM has been characterized and tested many times before. Further, identification and preparation of the hAM are unambiguous and straightforward (Jerman et al, 2014;Poženel et al, 2019;Jerman et al, 2020;Ramuta et al, 2020;Weidinger et al, 2020), whereas cells and cell-derived secretomes or sEV require precise characterization. Since the vast majority of the researchers isolated cells from the perinatal tissues by themselves, adequate characterization is mandatory to assure that a pure cell population has been isolated.…”