Stepwise Differentiation of Engineered Human Meniscus Tissues in 3D-Printed Flexible Scaffolds
Notice bibliographique
Résumé
Background Meniscus injuries are common with approximately 850,000 patients undergoing surgical treatment in the United States yearly. To investigate regenerative strategies for avascular meniscus tears, our lab has been utilizing bovine meniscus explants for investigating regenerative strategies for tears and crosstalk between joint tissues. However, inter-species crosstalk between bovine meniscus and human derived joint tissues remains a concern. This study was designed to engineer and validate a model of human meniscus for engineered healing of avascular meniscus tears under the crosstalk between meniscus, synovial membrane, and IFP. Methods A wedge-shaped scaffold was 3D-printed with Ecoflex F Blend C1200 polymer. Synovial mesenchymal stem/progenitor cells (syMSCs) were isolated from surgically removed synovium of anonymous adult donors as per our previous methods. The scaffolds were then loaded with collagen bioink and 2,000,000 cell /mL syMSCs and then placed into the PDMS molds. Experiment 1: For the first 2 weeks, the tissue constructs were cultured in Fibrogenic induction supplemented (FIS) media with connective tissue growth factor (CTGF). For weeks 3-4, the tissue constructs were cultured in chondrogenic induction supplemented media (CIS) and transforming growth factor beta-3 (TGFβ-3). Experiment 2: For 4 weeks tissue constructs were cultured in FIS with CTGF and CIS with TGFβ-3. No loading was applied. Experiment 3: Tissue constructs were cultured in combined FIS (w/ CTGF) and CIS (w/ TGFβ-3) with no FBS and no loading. Results At 4 weeks, histology showed new tissue integration into the pores throughout the scaffolds. Picrosirius Red (PR) and Alcian Blue (AB) staining suggest the de novo tissues in the no loading group are dense fibrocartilaginous tissues. Polarized images of PR-stained section showed densely aligned collagen fibers, reminiscent of native meniscus, comprising circumferential fibers and radial tie fibers. Mechanical loading resulted in less mature fibrous tissue matrix with limited cartilaginous matrix. Combined culture with FIS (w/ CTGF) and CIS (w/ TGFβ-3) showed the most mature and dense fibrocartilage tissue development after 4 weeks. RNA sequencing data show similar expression levels of fibrochondrocyte-specific genes, including COL1A2, ACAN, COMP, and COL1A1, between engineered and native human meniscus tissue. Conclusion Our study suggests a reliable and effective approach to engineer human meniscus-like tissues that can serve as an experimental explant model for various types of meniscus explant research. Combined culture with FIS and CIS creates more mature fibrocartilage structure than step-wise differentiation under no physiological loading. These findings may have implications for engineering human meniscus-like tissue for various types of meniscus research.
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 machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 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 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,001 | 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 source (Gemma direct ou Codex distillé), 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 ».