Organotypic and 3D Reconstructed Cultures of the Human Bladder and Urinary Tract

ABSTRACT Recent clinical studies suggest that the deliberate colonization of the human bladder with a prototypic asymptomatic bacteriuria-associated bacterium, Escherichia coli 83972, may reduce the frequency of urinary tract infection in individuals with spinal cord injuries. However, the mechanism by which E. coli 83972 colonizes the bladder is unknown. We examined the role in bladder colonization of the E. coli 83972 genes papG and fimH, which respectively encode P and type 1 receptor-specific fimbrial adhesins. E. coli 83972 and isogenic papGΔ and papGΔ fimHΔ mutants of E. coli 83972 were compared for their capacities to colonize the neurogenic human bladder. Both strains were capable of stable colonization of the bladder. The results indicated that type 1 class-specific adherence and P class-specific adherence, while implicated as significant colonization factors in experiments that employed various animal model systems, were not required for colonization of the neurogenic bladder in human beings. The implications of these results with regard to the selection of potential vaccine antigens for the prevention of urinary tract infection are discussed.

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Molecular Analysis of Asymptomatic Bacteriuria Escherichia coli Strain VR50 Reveals Adaptation to the Urinary Tract by Gene Acquisition

Infection and Immunity ◽

10.1128/iai.02810-14 ◽

2015 ◽

Vol 83 (5) ◽

pp. 1749-1764 ◽

Cited By ~ 13

Author(s):

Scott A. Beatson ◽

Nouri L. Ben Zakour ◽

Makrina Totsika ◽

Brian M. Forde ◽

Rebecca E. Watts ◽

...

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Mouse Model ◽

Molecular Mechanisms ◽

Asymptomatic Bacteriuria ◽

Human Bladder ◽

Content Type ◽

E Coli ◽

Tract Infections ◽

K 12

Urinary tract infections (UTIs) are among the most common infectious diseases of humans, withEscherichia coliresponsible for >80% of all cases. One extreme of UTI is asymptomatic bacteriuria (ABU), which occurs as an asymptomatic carrier state that resembles commensalism. To understand the evolution and molecular mechanisms that underpin ABU, the genome of the ABUE. colistrain VR50 was sequenced. Analysis of the complete genome indicated that it most resemblesE. coliK-12, with the addition of a 94-kb genomic island (GI-VR50-pheV), eight prophages, and multiple plasmids. GI-VR50-pheVhas a mosaic structure and contains genes encoding a number of UTI-associated virulence factors, namely, Afa (afimbrial adhesin), two autotransporter proteins (Ag43 and Sat), and aerobactin. We demonstrated that the presence of this island in VR50 confers its ability to colonize the murine bladder, as a VR50 mutant with GI-VR50-pheVdeleted was attenuated in a mouse model of UTIin vivo. We established that Afa is the island-encoded factor responsible for this phenotype using two independent deletion (Afa operon and AfaE adhesin) mutants.E. coliVR50afaand VR50afaEdisplayed significantly decreased ability to adhere to human bladder epithelial cells. In the mouse model of UTI, VR50afaand VR50afaEdisplayed reduced bladder colonization compared to wild-type VR50, similar to the colonization level of the GI-VR50-pheVmutant. Our study suggests thatE. coliVR50 is a commensal-like strain that has acquired fitness factors that facilitate colonization of the human bladder.

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An encapsulated drug delivery system for recalcitrant urinary tract infection

Journal of The Royal Society Interface ◽

10.1098/rsif.2013.0747 ◽

2013 ◽

Vol 10 (89) ◽

pp. 20130747 ◽

Cited By ~ 9

Author(s):

Sheyda Labbaf ◽

Harry Horsley ◽

Ming-Wei Chang ◽

Eleanor Stride ◽

James Malone-Lee ◽

...

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Slow Release ◽

Polymeric Carrier ◽

Human Bladder ◽

Fluorescent Tracer ◽

Bladder Cells ◽

Uropathogenic Bacteria

One of the hallmarks of urinary tract infection, a serious global disease, is its tendency to recur. Uropathogenic bacteria can invade cells lining the bladder, where they form longer-term intracellular reservoirs shielded from antibiotics, re-emerging at a later date to initiate flare-ups. In these cases, only lengthy systemic antibiotic treatment can eradicate all the reservoirs. Yet, long courses of antibiotics are not ideal, as they can lead to side effects and an increase in antibiotic resistance. Moreover, most antibiotics lose some potency by the time they reach the bladder, and many cannot permeate cells, so they cannot access intracellular reservoirs. Here, using coaxial electrohydrodynamic forming, we developed novel core–shell capsules containing antibiotics as a prototype for a future product that could be infused directly into the bladder. Gentamicin was encapsulated in a polymeric carrier (polymethylsilsesquioxane) and these capsules killed Enterococcus faecalis , a common chronic uropathogen, in vitro in a dose-responsive, slow-release manner. Capsules containing a fluorescent tracer dye in place of gentamicin penetrated human bladder cells and released their dye cargo with no apparent toxicity, confirming their ability to successfully permeate cells. These results suggest that such antibiotic capsules could prove useful in the treatment of recalcitrant UTI.

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Palmitate Induced IL-6 and MCP-1 Expression in Human Bladder Smooth Muscle Cells Provides a Link between Diabetes and Urinary Tract Infections

PLoS ONE ◽

10.1371/journal.pone.0010882 ◽

2010 ◽

Vol 5 (5) ◽

pp. e10882 ◽

Cited By ~ 30

Author(s):

Andreas Oberbach ◽

Nadine Schlichting ◽

Matthias Blüher ◽

Peter Kovacs ◽

Holger Till ◽

...

Keyword(s):

Smooth Muscle ◽

Urinary Tract ◽

Smooth Muscle Cells ◽

Urinary Tract Infections ◽

Muscle Cells ◽

Bladder Smooth Muscle ◽

Human Bladder ◽

Tract Infections

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Dynamic persistence of intracellular bacterial communities of uropathogenic Escherichia coli in a human bladder-chip model of urinary tract infections

10.1101/2021.01.03.424836 ◽

2021 ◽

Author(s):

Kunal Sharma ◽

Neeraj Dhar ◽

Vivek V. Thacker ◽

Thomas Simonet ◽

François Signorino-Gelo ◽

...

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Epithelial Cells ◽

Urinary Tract Infections ◽

Bacterial Communities ◽

Antibiotic Administration ◽

Human Bladder ◽

Tract Infections ◽

Intracellular Bacterial Communities

AbstractUropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infections and is a major cause for antibiotic prescriptions. Previous studies have shown that infection of terminally differentiated urinary bladder cells leads to the formation of intracellular bacterial communities (IBCs). However, the precise role of IBCs in recurrence of infection and antibiotic persistence, is not completely understood in part because the in situ dynamic responses of bacteria within these structures to antibiotic stress is difficult to assess in animal models. Here, we develop and characterize a human bladder-chip model of UPEC infection wherein superficial bladder epithelial cells and bladder microvascular endothelial cells are co-cultured under flow in urine and nutritive media respectively, and the mechanics of bladder filling and voiding cycles mimicked by application and release of linear strain. Time-lapse microscopy showed that infection of epithelial cells under shear stress in diluted urine led to the rapid recruitment and diapedesis of neutrophils across the endothelial-epithelial barrier and the formation of neutrophil swarms and neutrophil extracellular traps. Subsequently, two cycles of antibiotic administration interspersed with recovery periods revealed both non-growing and rapidly proliferating IBCs. Multiple stages of IBC formation captured on-chip with single-cell resolution revealed that that bacterial killing within IBCs was substantially delayed, outcomes such as shedding of bacteria and exfoliation are not mutually exclusive and rapidly reseeded the infection, and in rare instances bacterial growth in IBCs continued for the entire period of antibiotic administration. These new insights into the early stages of pathogenesis revisit the role of IBCs as harbours of persistent bacterial populations, with significant consequences for non-compliance with antibiotic regimens.

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Dynamic persistence of UPEC intracellular bacterial communities in a human bladder-chip model of urinary tract infection

eLife ◽

10.7554/elife.66481 ◽

2021 ◽

Vol 10 ◽

Author(s):

Kunal Sharma ◽

Neeraj Dhar ◽

Vivek V Thacker ◽

Thomas M Simonet ◽

Francois Signorino-Gelo ◽

...

Keyword(s):

Urinary Tract ◽

Bacterial Communities ◽

Recovery Period ◽

Dynamic Responses ◽

Neutrophil Extracellular Trap ◽

Antibiotic Administration ◽

Human Bladder ◽

Tract Infections ◽

Intracellular Bacterial Communities ◽

Umbrella Cells

Uropathogenic Escherichia coli (UPEC) proliferate within superficial bladder umbrella cells to form intracellular bacterial communities (IBCs) during early stages of urinary tract infections. However, the dynamic responses of IBCs to host stresses and antibiotic therapy are difficult to assess in situ. We develop a human bladder-chip model wherein umbrella cells and bladder microvascular endothelial cells are co-cultured under flow in urine and nutritive media respectively, and bladder filling and voiding mimicked mechanically by application and release of linear strain. Using time-lapse microscopy, we show that rapid recruitment of neutrophils from the vascular channel to sites of infection leads to swarm and neutrophil extracellular trap formation but does not prevent IBC formation. Subsequently, we tracked bacterial growth dynamics in individual IBCs through two cycles of antibiotic administration interspersed with recovery periods which revealed that the elimination of bacteria within IBCs by the antibiotic was delayed, and in some instances, did not occur at all. During the recovery period, rapid proliferation in a significant fraction of IBCs reseeded new foci of infection through bacterial shedding and host cell exfoliation. These insights reinforce a dynamic role for IBCs as harbours of bacterial persistence, with significant consequences for non-compliance with antibiotic regimens.

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Scanning Electron Microscopy of Human Bladder Mucosa in Acute and Chronic Urinary Tract Infection

British Journal of Urology ◽

10.1111/j.1464-410x.1984.tb07160.x ◽

1984 ◽

Vol 56 (1) ◽

pp. 38-43 ◽

Cited By ~ 22

Author(s):

T. S. J. ELLIOTT ◽

R. C. B. SLACK ◽

M. C. BISHOP

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Bladder Mucosa ◽

Human Bladder ◽

Chronic Urinary Tract Infection ◽

Scanning Electron

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Transurethral resection of a bladder leiomyoma: A case report

Canadian Urological Association Journal ◽

10.5489/cuaj.1335 ◽

2014 ◽

Vol 8 (1-2) ◽

pp. 111 ◽

Cited By ~ 3

Author(s):

Goksel Hasan Goktug ◽

Ufuk Ozturk ◽

Nevzat Can Sener ◽

Can Tuygun ◽

Hasan Bakirtas ◽

...

Keyword(s):

Case Report ◽

Urinary Tract ◽

Magnetic Resonance ◽

Transurethral Resection ◽

Disease Recurrence ◽

Malignant Tumours ◽

Human Bladder ◽

Tract Infections ◽

Bladder Leiomyoma ◽

Bladder Mass

Benign mesenchimal tumour of the human bladder is rare. Insulin potentiation therapy mimics malignant tumours both clinically and radiologically. We present a patient we treated with transurethral resection (TUR) only. A 27-year old male patient presented to our clinic with frequency, dysuria and recurrent urinary tract infections. Magnetic resonance (MRI) revealed an endovesical bladder mass of 7 × 8 cm. We performed TUR in the same session for both diagnosis and treatment. The diagnosis was endovesical leiomyoma. Six months to a year after the operation, the MRI did not reveal disease recurrence. Even though TUR is recommended for smaller and endovesical tumours, we believe larger intravesical tumours may also be managed by TUR.

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