This study aimed to find an optimal formulation to form 3D hyaline-like cartilage substitutes using the tissue engineering triads. The primary cells taken from osteoarthritic patients were overexpressed with transcriptional factor SRY (Sex Determining Region Y)-Box 9 (SOX9) using Lipofectamine 2000™ through a non-viral transfection method. The transfected and non-transfected cells were seeded on poly(lactic-co-glycolic acid) (PLGA) based scaffolds with and without fibrin. The arrangement resulted in four experimental groups. The 3D ‘cells-scaffolds’ tissue constructs were cultured for three weeks and implanted ectopically in nude mice for four weeks. The evaluations include macroscopic and microscopic study, gene expression analyses, and sulfated glycosaminoglycan (sGAG) assay, focusing on the cartilage properties. A biomechanical evaluation was performed only on post-implanted constructs. All in vitro, two- and four-week post-implanted constructs exhibited firm and smooth hyaline-like cartilage appearance. In vitro constructs showed sparse cells distribution with minimal cartilaginous tissue formation. However, a high density, lacunae-encapsulated chondrocytes embedded within the basophilic ground substances was observed in all post-implanted constructs. It is supported by positive-brownish precipitation immunolocalisation against collagen type II. Besides, molecular analysis showed that COL2A1 and other cartilaginous markers were also expressed. Increased sGAG content and compressive strain could be observed in vitro and in vivo. Although quantitatively, no significant statistical differences were found between the four groups, the qualitative results indicated that SOX9-overexpressed cells, PLGA, and fibrin combination guides hyaline-like cartilage formation better than other groups. Hence, the combination may be studied in a big animal model to develop its potential for future clinical application.
Articular cartilage has poor repair capacity due to its avascular and aneural properties and has relatively few cells. This study investigated the ability of autologous implantation approach using three dimensional (3D) constructs engineered from bone marrow mesenchymal stem cells (BMSCs) seeded on poly(lactic-co-glycolic acid) (PLGA) with or without fibrin as cells carrier for the repair of osteochondral defect in rabbit model. The engineered 3D constructs – PLGA/Fibrin/BMSCs and PLGA/BMSCs – were cultured for 3 weeks in vitro and implanted autologously to the osteochondral defect created in the rabbit knee. The in vivo constructs were harvested and evaluated by means of gross observation, histology assessment, gene expression study, sulphated glycosaminoglycan (sGAG) production assay and biomechanical evaluation at 6 and 12 weeks post implantation. The results showed that the osteochondral defects treated with the PLGA/Fibrin/BMSCs constructs exhibited better repairment, more cartilaginous extracellular matrix, higher sGAG production, superior compressive strength and more intense expression of chondrogenic marker genes than the PLGA/BMSCs group. This study suggested that the PLGA/Fibrin/BMSCs has the potential to treat osteochondral defect and may be presented as a viable therapeutic option for those who would be in need from the life-extending benefits of tissue replacement or repair.
Introduction: Treating damaged articular cartilage is a predicament in the orthopaedic field due to its limited capacity for regeneration. Efforts to find the best available treatment for articular cartilage injuries are still actively ongoing. This study investigates the ability of the autologous implantation approach using three dimensional (3D) constructs engineered from bone marrow mesenchymal stem cells (BMSCs) seeded on poly(lactic-co-glycolic acid) (PLGA) with or without fibrin as cells carrier for the repair of full-thickness osteochondral defect in rabbit model. Methods: 3D PLGA was fabricated via solvent casting-salt leaching technique using salt as porogen. The BMSCs obtained from the New Zealand white rabbits were cultured and mixed with (1) autologous plasma-derived fibrin or, (2) culture media which served as the control, prior to ‘cells seeding’ procedure into the PLGA. After three weeks of in vitro culture, the engineered 3D constructs - PLGA/Fibrin/BMSCs (PFB) and PLGA/BMSCs (PB) - were implanted autologously to the osteochondral defect created in the rabbit’s knee. The in vivo constructs were harvested en bloc and evaluated by gross inspection and histology, gene expression, sulphated glycosaminoglycan (sGAG) production and biomechanical defect property at 6 and 12 weeks post implantation (n=6 for each group). Results: The results showed that the osteochondral defects treated with the PFB constructs exhibited better repairment, more cartilaginous extracellular matrix, higher sGAG content, significantly stronger compressive strength and greater expression of chondrogenic marker genes than the PB group. Conclusions: The findings suggest that PFB with optimal induction are feasible to treat osteochondral and may become a potential treatment modality aiming to treat articular cartilage disease in human.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.