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Dynamically Urethra-Adapted and Obligations-Oriented Trilayer Hydrogels Integrate Scarless Urethral Repair

ABSTRACT: In urethral damage/stricture prevention, open and harsh urethral microenvironments as well as the isotropic compression and swelling properties of exogenous implants render urethral repair intractable. Here, a dynamically urethra-adapted and obligation-oriented trilayer hydrogel was constructed for integrated scarless urethral repair. The bottom diethylacrylamide -hydroxyethylacrylamide (D-H) hydrogel featuring high anti-fouling performance, the upper swellable verteporfin (VP)-loaded N,N'-methylenebisacrylamide-poly (N-isopropylacrylamide) (BP) with a hydrophilic Janus surface promote urethral regeneration through expediting cell migration and proliferation and the rigid and water-resistant Zein middle layer opposes urine voiding-arised BP shedding and resists inward BP swelling-driven urethral occlusion and urine permeation. Importantly, systematic proteomic and genomic analyses revealed that such hydrogel scaffolds expedite regeneration, alleviate inflammation and regulate extracellular matrix secretion, where BP swelling-induced outgrowth for fibrosis and scar formation is significantly inhibited by VP through the blockade of the scar-associated YAP signaling pathway. These microenvironment-adaptable design concepts provide a rationale for engineering urethral regeneration scaffolds.

ORGANISM(S): Oryctolagus cuniculus

PROVIDER: GSE301213 | GEO | 2025/07/01

REPOSITORIES: GEO

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Project description:Background: Urethral stricture is related to scar tissue fibrosis, but its pathogenesis is still unclear. This study aims to explore the mechanism of circular RNA (circRNA) in the occurrence and development of urethral stricture. Methods: CircRNA microarray was employed to analyze circRNA expression profiles between human urethral scar tissue and normal urethral tissue. The first nine circRNA differentially expressed were selected for RT-qPCR detection. Urethral scar fibroblasts were isolated from human urethral scar tissue and cultured in vitro. The cells were transfected with si-circ_0047339, LV-circ_0047339, and miR-4691-5p mimic and their corresponding negative controls. Related gene and protein expression was measured by RT-qPCR, Western Blot and immunofluorescence staining, and the cell function was detected by CCK-8 and EDU assay. The targeting relationship between miR-4691-5p and circ_0047339 or TSP-1 was analyzed by dual-luciferase reporter assay. Results: The results of circRNA microarray showed that there were 268 differential genes between urethral scar tissue and normal urethral tissue (circRNA expression difference multiple ≥2 and p value≤0.05). The enrichment analysis of KEGG (Kyoto encyclopedia of genes and genomes) showed that these circRNAs were significantly correlated with ECM-receptor interaction. The first nine differentially expressed circRNA were selected to predict the circRNA-miRNA network. RT-qPCR results showed that circ_0047339 was upregulated considerably in urethral scar tissue. After silencing circ_0047339, the proliferation of urethral scar cells decreased significantly, and the expression of collagen 1 (COL-1) and α-smooth muscle actin (α-SMA) also reduced. Circ_0047339 as a competing endogenous RNA (ceRNA) could increase the expression of TSP-1 by competitively binding miR-4691-5p. In addition, miR-4691-5p mimic transfection could inhibit the proliferation of urethral scar fibroblasts and the presentation of thrombospondin-1 (TSP-1), α-SMA and COL-1, while circ_0047339 overexpression eliminated this inhibition. Conclusion: Our results showed that the dysfunctional circ_0047339 might promote the growth and fibrosis of urethral scar fibroblasts through the ceRNA mechanism, thus promoting the development of urethral stricture via miR-4691-5p/TSP-1 axis.

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Dynamically Urethra-Adapted and Obligations-Oriented Trilayer Hydrogels Integrate Scarless Urethral Repair

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