(006) ADVANCING PD.SPHERE: ENHANCED PROTOCOLS FOR A MORPHOLOGICALLY AND PATHOLOGICALLY ACCURATE 3D CELLULAR MODEL IN PEYRONIE’S DISEASE RESEARCH
Notice bibliographique
Résumé
Abstract Introduction Peyronie's disease (PD) affects about 10% of men and is often studied using traditional monolayer fibroblast monolayer cultures and limited animal models, which do not adequately replicate the complex cellular interactions of PD. We have developed an advanced three-dimensional (3D) cellular model named PD.SPHERE using a novel 'micro-gravity' cell culture technique from human PD tissues. Significant advancements in the culturing protocol have improved the morphology and physiopathology of these models, providing an essential platform for in vitro treatment screening. Objective The aim is to refine the culturing protocol for generating PD.SPHEREs that more accurately mimics the morphological and physiological features of PD plaques and to assess the efficacy of these refined models as tools for treatment screening. Methods Fibroblasts from human PD tissues were isolated and cultured using the 'micro-gravity' technique with modifications. The PD fibroblast cells were cultured and propagated in a non-adhesive 96-well plate, with and without 10 ng/ml transforming growth factor-beta (TGF-β) supplementation, under micro-gravity conditions. The plates were then placed on an orbital shaker inside the incubator and incubated at 37°C with 5% CO2 and 95% relative humidity. We confirmed the appropriate morphology and cellular compositions, including α-smooth muscle actin (α-SMA) and cellular collagen, using microscopic assays. The refined 3D model was subsequently used to evaluate the effects of collagenase from Clostridium histolyticum (CCH) and actinidin, a collagenase-like enzyme derived from kiwi fruits. Results Our findings from the refined 3D model revealed significant changes in cellular morphology. Specifically, PD.SPHEREs developed from cells that were not initially supplemented with TGF-β, but were later cultured in micro-gravity with TGF-β supplementation, and PD.SPHEREs from cells both propagated and cultured with TGF-β were approximately 2.3 and 3.5 times larger, respectively, than those PD.SPHEREs propagated and cultured entirely without TGF-β (P < 0.0001). The collagen content was significantly highest (P < 0.01) in PD.SPHEREs that were both propagated and cultured with TGF-β, compared to all other groups. The refined model was susceptible to both CCH and actinidin. The size and collagen content of the refined PD.SPHEREs significantly decreased (P < 0.05) following treatment with CCH and actinidin. Conclusions The advancements made in the culturing protocol of PD.SPHEREs represent a significant improvement in the modeling of PD for in vitro studies. Our results demonstrate the critical role of TGF-β in the growth and morphology of PD.SPHEREs. The increase in sized collagen content of the spheroids when cultured in the presence of TGF-β suggests that this factor plays a substantial role in cellular proliferation and cellular matrix accumulation, which are key aspects of PD pathogenesis. Furthermore, the increased collagen content in PD.SPHEREs cultured with TGF-β aligns with the known pathophysiology of PD, where collagen accumulation contributes to plaque formation and disease progression. This finding underscores the utility of our model in replicating key pathological features of PD and supports the potential of PD.SPHERE as an effective tool for screening and evaluating the efficacy of treatments aimed at modulating these pathological processes. Disclosure No.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
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 | 0,005 | 0,003 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,001 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| 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 | 0,001 |
| 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 ».