(237) INVESTIGATING THE EFFECTS OF ACTINIDIN AS A COLLAGENASE ON HUMAN PEYRONIE’S DISEASE CELLS

Abstract Introduction Peyronie's Disease (PD) is a condition characterized by a connective tissue disorder that affects the penile tunica albuginea, leading to the formation of collagen-rich plaques. These plaques can cause penile curvature, painful erections, and erectile dysfunction. With the withdrawal of Collagenase C. histolyticum injections from the Canadian market, treatment options for PD have become limited. Actinidin, a soluble enzyme capable of breaking down various forms of collagen and fibrinogen, has shown effectiveness in reducing collagen in human in vitro cell models. Objective To assess the cytotoxicity of actinidin and evaluate its efficacy in reducing intercellular bound collagen in a 2D cellular human Peyronie's disease plaque models. Methods A 2D cellular model was cultured using human fibroblast cells isolated from PD tissues (N ≥ 4). Actinidin treatment was prepared using isolated actinidin powder in media, filtered, and adjusted to pH 7. Cells were treated with 10 ng/ml of TGF-β to induce a PD-like cellular model for 24 hours and then exposed to actinidin concentrations ranging from 1 mg/ml to 20 mg/ml. Media-only and TGF-β-only controls were included. Cell proliferation was assessed at 24 hours at various actinidin concentrations using an MTT assay. Collagen I levels were quantified through western blot analysis after 24 hours of treatment. Results were analyzed using oneway Anova analysis. Results Results from both the cell proliferation and collagen I quantification revealed that TGF-b significantly increases cell proliferation (P = 0.042) and collagen I levels (P = 0.018). After treating the cells with various concentrations of actinidin, the cell proliferation assay revealed that cell viability at 20 mg/ml was significantly lower (P = 0.038) compared to the TGF-β control. Additionally, a trend was observed where cell proliferation appeared to slightly decrease at higher concentrations of actinidin, although there was no significant difference in cell viability between the other treated concentrations and the TGF-β control. Western blot analysis showed that Collagen I levels were significantly reduced after 24 hours of actinidin treatment at concentrations of 10 mg/ml and 20 mg/ml. Conclusions Our continuation preliminary results demonstrate that actinidin can reduce collagen I levels in PD cells. However, high doses of actinidin cause cell destruction, so we recommend that treatment should not exceed 20 mg/ml. Future studies will investigate the molecular mechanism by which actinidin hydrolyzes collagen and will evaluate its effects in an animal model. Given the lack of durable FDA-approved treatments for PD in Canada, this novel approach holds significant potential for future treatment options. Disclosure Yes, this is sponsored by industry/sponsor: KiwiEnzyme.com Ltd. Clarification: No industry support in study design or execution.