Precision-Cut Bladder Slices: A Novel Model for the Study of Bladder Fibrosis and Potential Anti-Fibrotic Agents
Yutao Lu1, Natasja Patricia Simonsen1, Jens Christian Djurhuus1
1Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
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.
