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INTRODUCTION: Bladder cancer ranks among the top ten most frequently diagnosed cancers globally, with approximately 25% of diagnosed cases presenting as initially aggressive muscle-invasive tumours, marked by poor prognosis and frequent metastasis. Human amniotic membrane (hAM) is an extraembryonic membrane with antiinflammatory, antifibrotic and antimicrobial properties, however, despite the growing number of studies, the cellular mechanisms underpinning its anticancer activity remain poorly elucidated. In our investigation, we delved into the mechanisms by which hAM homogenate influences bladder cancer and normal urothelial cells. MATERIALS AND METHODS: The hAM homogenate was prepared from placentas collected from healthy donors after elective caesarean section according to the protocol described in. Wound healing assay was used to evaluate the influence of hAM homogenate on migration of normal porcine urothelial NPU cells, human papillary cancer urothelial RT4 cells and human muscle-invasive bladder cancer T24 cells. For the invasion of cancer cells, invasion assay was performed using transwell chambers with membranes of 8 μm pore size coated with diluted Matrigel matrix. The effect of hAM homogenate on the migration pattern of T24 cells stably transfected with enhanced green fluorescent protein (eGFP) was examined using time-lapse confocal microscopy. Protein expression levels of molecules involved in cell migration were measured by Western blot analysis, while gene expression was quantified by RT-qPCR. To compare statistical differences between at two experimental groups, parametric unpaired two-tailed Student’s t-test or the nonparametric Mann–Whitney test was used. RESULTS AND DISCUSSION: Our study revealed that hAM homogenate impedes the migration rate of bladder cancer RT4 and T24 cells, but not of normal NPU cells. In addition to cell migration, hAM homogenate significantly decreased the invasion rate of muscle-invasive bladder cancer T24 cells. Furthermore, this antimigratory effect was associated with the downregulation of FAK and Rho GTPases expression—crucial proteins in actin cytoskeleton reorganization. CONCLUSIONS: In summary, the findings highlight the significant potential of hAM homogenate as a valuable biomaterial in the field of medicine, particularly as a complementary component in the treatment of bladder cancer. [ABSTRACT FROM AUTHOR]

Urinary tract infections (UTIs) represent a serious global health issue, especially due to emerging multidrug-resistant UTI-causing bacteria. Recently, we showed that the human amniotic membrane (hAM) could be a candidate for treatments and prevention of UPEC and Staphylococcus aureus infections. However, its role against multidrug-resistant bacteria, namely methicillin-resistant S. aureus (MRSA), extended-spectrum beta-lactamases (ESBL) producing Escherichia coli and Klebsiella pneumoniae, vancomycin-resistant Enterococci (VRE), carbapenem-resistant Acinetobacter baumannii, and Pseudomonas aeruginosa has not yet been thoroughly explored. Here, we demonstrate for the first time that the hAM homogenate had antibacterial activity against 7 out of 11 tested multidrug-resistant strains, the greatest effect was on MRSA. Using novel approaches, its activity against MRSA was further evaluated in a complex microenvironment of normal and cancerous urinary bladder urothelia. Even short-term incubation in hAM homogenate significantly decreased the number of bacteria in MRSA-infected urothelial models, while it did not affect the viability, number, and ultrastructure of urothelial cells. The hAM patches had no antibacterial activity against any of the tested strains, which further exposes the importance of the hAM preparation. Our study substantially contributes to basic knowledge on the antibacterial activity of hAM and reveals its potential to be used as an antibacterial agent against multidrug-resistant bacteria. [ABSTRACT FROM AUTHOR]