MP66-02 INVESTIGATING ACTINIDIN AS A NOVEL COLLAGENASE ON HUMAN PEYRONIE’s DISEASE CELLS

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You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology (MP66)1 May 2024MP66-02 INVESTIGATING ACTINIDIN AS A NOVEL COLLAGENASE ON HUMAN PEYRONIE's DISEASE CELLS Kevin Feng, Wongsakorn Kiattiburut, Jeremy Burton, and Jeffrey D. Campbell Kevin FengKevin Feng , Wongsakorn KiattiburutWongsakorn Kiattiburut , Jeremy BurtonJeremy Burton , and Jeffrey D. CampbellJeffrey D. Campbell View All Author Informationhttps://doi.org/10.1097/01.JU.0001009468.01097.19.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Peyronie's Disease (PD) is a connective tissue disorder in the penile tunica albuginea characterized by the formation of collagen rich plaques. Intralesional Collagenase Clostridium histolyticum injections are no longer available in Canada, which limits treatment options for PD. Actinidin, a soluble enzyme, possesses the capability to hydrolyze various types of collagens and fibrinogen. Our primary goal is to assess the dose-response efficacy and cytotoxicity of actinidin in reducing intercellular-bound collagen in human PD plaque models. METHODS: Human PD fibroblast cells were isolated and cultured in 2D models. Various treatment groups were administered, including media-only control and actinidin concentrations ranging from 0.5 mg/ml to 30 mg/ml. Actinidin was isolated from Hayward kiwi by filtration and adjustment to 7.0 pH. Toxicity was assessed using a MTT assay to generate a dose-response curve at multiple time points and concentrations. Collagen quantification and expression were measured using Soluble Collagen Quantification Assay Kit and Collagen I antibody after 24 hours of treatment. Exploration of extracellular matrix effects involved cell staining for the nucleus and actin filaments, followed by a twelve-hour fluorescent confocal microscope timelapse capturing fluorescein images. The total fluorescent area of each well was measured, and actin signals were normalized using nucleus signals. RESULTS: The dose-response curve revealed that the lethal concentration affecting 50% of the cells after a 48-hour period is approximately 15-20 mg/ml. Western blot and collagen quantification show that treatment concentrations of greater than 10 mg/mL actinidin significantly reduced cellular collagen of human PD fibroblasts compared to the control group (p=0.024). The high concentration actinidin treatment (30 mg/mL) exhibited a significantly higher normalized actin signal compared to other treatment groups (p<0.001). These findings suggest that actinidin may possess mechanisms that affect the plasma membrane integrity. CONCLUSIONS: Our ongoing investigations reveal actinidin's capacity to degrade PD cells by affecting extracellular collagen and cellular membrane. We recommend not exceeding 15 mg/ml as it will actinidin doses result in cell destruction. Future research will continue to delve into the molecular mechanisms of actinidin in collagen hydrolysis and assess its impact in an animal model. In light of the absence of enduring FDA-approved PD treatments in Canada, this novel approach holds promise for future therapeutic potential. Source of Funding: Canadian Urology Association Scholarship Foundation; The Lavergne Catalyst Grant (Department of Urology, Western University) © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1086 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Kevin Feng More articles by this author Wongsakorn Kiattiburut More articles by this author Jeremy Burton More articles by this author Jeffrey D. Campbell More articles by this author Expand All Advertisement PDF downloadLoading ...

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