Precision‐Cut Bladder Slices: A Novel Model for the Study of Bladder Fibrosis and Potential Anti‐Fibrotic Agents

ABSTRACT

Objectives
Precision‐cut tissue slice culture is an innovative ex vivo approach for studying fibrosis pathogenesis. Here, we report for the first time the use of human precision‐cut bladder slices (PCBS) to investigate fibrotic changes and evaluate anti‐fibrotic compounds.


Methods

Fresh bladder tissue was obtained from 16 patients undergoing surgery for non‐fibrotic conditions and 7 patients with documented bladder fibrosis. PCBS were cultured and stimulated with transforming growth factor β (TGF‐β) to induce fibrotic changes. Viability was assessed by ATP quantification. The anti‐fibrotic efficacy of pirfenidone (PFD), relaxin‐2 (RLN), bone morphogenetic protein 7 (BMP‐7), imatinib (IMA), and galunisertib (GAL) was evaluated. Fibrosis was quantified using qPCR analysis of collagen 1 (
COL1A1
), fibronectin (
FN1
), and cellular communication network factor 2 (
CCN2
) gene expression.



Results

PCBS remained viable over 48 h, with stable ATP levels. TGF‐β stimulation significantly increased
COL1A1
and
CCN2
expression, confirming induction of a fibrotic response. Treatment with PFD, IMA, and GAL effectively attenuated TGF‐β‐induced upregulation of fibrosis markers. At baseline, PCBS derived from fibrotic bladders exhibited elevated
COL1A1
expression compared to non‐fibrotic tissue, while
FN
and
CCN2
levels remained unchanged. PFD treatment notably reduced
CCN2
expression in fibrotic PCBS.



Conclusions
This study demonstrates that PCBS provide a viable and reproducible platform for modeling bladder fibrosis and screening anti‐fibrotic therapies. PFD, IMA, and GAL showed promising anti‐fibrotic effects, supporting further investigation into their therapeutic potential.