(080) Exploring the Effects of Actinidin as a Novel Collagenase on Human Peyronie’s Disease Cells
Notice bibliographique
Résumé
Abstract Introduction Peyronie’s Disease (PD) is a connective tissue disorder of the penile tunica albuginea that affects approximately 10% of men. The plaques that hallmark PD contain excessive amounts of collagen and can result in penile curvature, painful erections, and/or erectile dysfunction. Intralesional Collagenase Clostridium histolyticum injection for PD are no longer available in Canada, making treatment options limited. Hayward kiwi fruit are rich in natural actinidin, a soluble enzyme that has the unique ability to hydrolyze various types of collagen and fibrinogen. Actinidin is considered a collagenase alternative; however, there have been no studies evaluating the application of actinidin on in vitro cellular PD models. Objective To determine the effectiveness of actinidin at reducing the collagen component of a 2D human PD cell model. Secondly, to determine the cytotoxicity of kiwifruit extract on human PD cells. Methods Human PD tissues obtained during penile prosthesis implantation were isolated for fibroblast cells and cultured using complete media. The cells were plated on a 96-well plate and treated with one of 4 treatment groups: media, saline, and low and high concentration actinidin treatment (0.5 mg/ml to 37.5 mg/ml). The actinidin treatments were prepared by dissolving freeze-dried kiwi powder in saline and then filtered; the low concentration was further diluted with saline. Cells were incubated at 37°C for 24 hours and then collagen was extracted and measured using a soluble collagen quantification assay kit and a fluorescent microplate reader. To measure cytotoxicity, cells were stained for the nucleus and actin filaments using DAPI and Phalloidin-conjugated FITC, respectively. A twelve-hour timelapse capturing images of FITC and DAPI fluorescein were carried through using a fluorescent confocal microscope. The total fluorescent area of each well was measured using ImageJ. Actin signals were normalized by DAPI signals, and all values were analyzed using Python and Prism10. Results The higher concentration of actinidin treatment significantly reduced cellular collagen of human PD fibroblasts compared to the media treatment (P < 0.05). The high concentration actinidin treatment had a significantly higher normalized actin signal than other treatment groups (P < 0.0001). This ratio allows us to observe the treated cells with compromised plasma membranes. The low concentration of actinidin did not have any significant difference from media and saline treatment groups. A significant increased level of normalized actin indicated that actinidin may have mechanisms to disturb the plasma membrane. Conclusions Our preliminary study suggests that the natural collagenase actinidin can break down PD cells by reducing the extracellular collagen content and compromising the cellular membrane. Further studies are underway to investigate treatment in 3D models and to determine the optimal actinidin concentration that can further reduce the collagen composition in human PD cell models. Actinidin has the potential to be a novel treatment option for patients suffering from PD. Disclosure Yes, this is sponsored by industry/sponsor: KiwiEnzyme.com Ltd Clarification No industry support in study design or execution
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,002 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,001 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».