Development of a Wound-Healing Protocol for In Vitro Evaluation of Urothelial Cell Growth

• Published in: Methods and protocols Vol. 6; no. 4; p. 64
• Main Authors: Foster, Christopher, Jensen, Todd, Finck, Christine, Rowe, Courtney K
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 05.07.2023; MDPI
• Subjects: Angiogenesis; Bladder; Cell growth; Comparative analysis; Cornea; Epidermal growth factor; Ethylenediaminetetraacetic acid; Fibroblast growth factor 2; Growth; growth factor; Growth factors; Health aspects; Insulin-like growth factor I; Insulin-like growth factors; Platelet-derived growth factor; Prostate; Quality of life; Regenerative medicine; Scientific equipment and supplies industry; Statistical analysis; Surgery; Transforming growth factor-a; Transforming growth factor-b1; Trauma; urethra; Vascular endothelial growth factor; Wound healing; United States; Connecticut; United States > US; growth factor; wound healing; urethra; regenerative medicine
• ISSN: 2409-9279; 2409-9279
• DOI: 10.3390/mps6040064

Urethral healing is plagued by strictures, impacting quality of life and medical costs. Various growth factors (GFs) have shown promise as therapeutic approaches to improve healing, but there is no protocol for in vitro comparison between GFs. This study focuses the development of a biomimetic in vitro urothelial healing assay designed to mimic early in vivo healing, followed by an evaluation of urothelial cell growth in response to GFs. Wound-healing assays were developed with human urothelial cells and used to compared six GFs (EGF, FGF-2, IGF-1, PDGF, TGF-β1, and VEGF) at three concentrations (1 ng/mL, 10 ng/mL, and 100 ng/mL) over a 48 h period. A commercial GF-containing medium (EGF, TGF-α, KGF, and Extract P) and a GF-free medium were used as controls. There was a statistically significant increase in cell growth for IGF-1 at 10 and 100 ng/mL compared to both controls ( < 0.05). There was a statistically significant increase in cell growth for EGF at all concentrations compared to the GF-free medium control ( < 0.05). This study shows the development of a clinically relevant wound-healing assay to evaluate urothelial cell growth. It is the first to compare GFs for future use in reconstructive techniques to improve urethral healing.