A Biomimetic Porcine Urothelial Model for Assessing Escherichia coli Pathogenicity

Abstract

Urinary tract infections can be severe, sometimes fatal, diseases whose etiological pathogens are predominantly uropathogenic strains of E. coli (UPEC). To investigate the UPEC pathogenesis, several models have already been established with minor or major disadvantages. The aim was to develop a simple, fast, and inexpensive biomimetic in vitro model based on normal porcine urothelial (NPU) cells that are genetically and physiologically similar to human bladder urothelium and to perform basic studies of E. coli pathogenicity. Initially, the model was tested using a set of control E. coli strains and, subsequently, with human E. coli strains isolated either from patients with urinary infections or from the feces of healthy individuals. A drop in viability of NPU cells was used as a measure of the pathogenicity of the individual strain tested. To visualize the subcellular events, transmission and scanning electron microscopy was performed. The strains were tested for the presence of different virulence-associated genes, phylogroup, type of core lipid, O-serotype, and type of lipopolysaccharide and a statistical analysis of possible correlations between strains’ characteristics and the effect on the model was performed. Results showed that our model has the discriminatory power to distinguish pathogenic from non-pathogenic E. coli strains, and to identify new, potentially pathogenic strains.

The Golgi complex: An organelle that determines urothelial cell biology in health and disease

“…We have found that more glycocalyx is present in these vesicles when UCs are not terminally differentiated, which is usually the case when UCs are grown in vitro (see Fig. 1C, F, K in the reference (Predojević et al 2022 ).…”

Molecular, morphological and functional properties of tunnelling nanotubes between normal and cancer urothelial cells: New insights from the in vitro model mimicking the situation after surgical removal of the urothelial tumor

“…In this study, we first examined spontaneously formed TNTs in monocultures (homocellular TNTs) of normal porcine urothelial (NPU) cells and invasive urothelial cancer T24 cells. The NPU cell model corresponded to normal human urothelial cells due to the structural and functional compatibility of porcine-derived NPU cells with normal human urothelial cells ( Visnjar et al, 2012 ; Resnik et al, 2015 ; Višnjar and Kreft, 2015 ; Resnik et al, 2018 ; Predojevic et al, 2022 ). Next, TNTs were studied in cocultures (heterocellular TNTs) of NPU and T24 cells to simulate intercellular communication in cancer tissues.…”

Methods and criteria for validating the multimodal functions of perinatal derivatives when used in oncological and antimicrobial applications

“…For example, a multilayered biomimetic porcine urothelial model that has been shown to react differently to pathogenic vs. non-pathogenic E. coli strains has been used for evaluating the antimicrobial properties of PnDs ( Ramuta T.Ž. et al, 2021 ; Predojević et al, 2022 ). Similar complex in vitro models have been established to study the host-pathogen interactions in the airway mucosa ( Marrazzo et al, 2016 ; Hasan et al, 2018 ), intestine ( Pearce et al, 2018 ; García-Díaz et al, 2022 ) and skin ( Bolle et al, 2020 ), but have not yet been used for evaluation of PnDs.…”