(133) Investigating Actinidin as a Potential Collagenase on Human Peyronie’s Disease Cells
Bibliographic record
Abstract
Abstract Introduction Peyronie's Disease (PD) is characterized by a connective tissue disorder in the penile tunica albuginea and the formation of collagen-rich plaques, leading to penile curvature, painful erections, and erectile dysfunction. Intralesional Collagenase Clostridium histolyticum injections are no longer available in Canada, making treatment options for PD limited. Actinidin is a soluble enzyme that has the ability to hydrolyze various types of collagen and fibrinogen. We have previously reported the efficacy of actinidin on reducing collagen in in vitro human PD models. Objective To determine the dose-response efficacy and cytotoxicity of actinidin in reducing the intercellular bound collagen in human PD plaque models. Methods Fibroblast cells are isolated from human PD tissue (N ≥ 3) and 2D models are cultured. These cells were subjected to various treatment groups including media-only control and 5 concentrations of actinidin treatment ranging from 0.5 mg/ml to 30 mg/ml. The collagenase treatment was prepared from Hayward kiwi by isolating actinidin by filtration and pH adjustment. A dose-response curve was generated at different time points at various actinidin concentrations using an MTT assay. After 24 hours of treatment, collagen extraction was performed and quantified using a Soluble Collagen Quantification Assay Kit and a fluorescent microplate reader. The effects of the treatments on the extracellular matrix were further investigated by staining the cells with DAPI for the nucleus and Phalloidin-conjugated FITC for actin filaments. A twelve-hour timelapse was conducted using a fluorescent confocal microscope to capture FITC and DAPI fluorescein images. The total fluorescent area of each well was measured, and actin signals were normalized by the DAPI 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. Treatment of 1 mg/mL actinidin significantly reduced cellular collagen of human PD fibroblasts compared to the control group (P < 0.001). The normalized stained β-actin signal was measured as the ratio between intracellular actin staining and nuclei count in each microscopic field. The high concentration actinidin treatment (30 mg/mL) exhibited a significantly higher normalized actin signal compared to other treatment groups (P < 0.0001). These findings suggest that actinidin may possess mechanisms that affect the plasma membrane integrity. Conclusions Our continuation of preliminary results demonstrates the ability of actinidin to break down PD cells by compromising extracellular collagen and the cellular membrane. High doses of actinidin cause cell destruction and we suggest that treatment should not exceed 10 mg/ml. Further studies will investigate the molecular mechanism of actinin in hydrolyzing collagen and to evaluate the effects on an animal model. Given the absence of durable FDA-approved treatments for PD in Canada, this novel option holds future treatment potential. Disclosure Yes, this is sponsored by industry/sponsor: KiwiEnzyme.com Ltd. Clarification: No industry support in study design or execution.
Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.
How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.002 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".