Cartilage tissue engineering by co-transplantation of chondrocyte extracellular vesicles and mesenchymal stem cells, entrapped in chitosan-hyaluronic acid hydrogel.

Mesenchymal stem cells (MSCs) on injectable hydrogels are mostly used to regenerate articular cartilage, which would have a variety of outcomes. Chondrocyte extracellular vesicles (EVs) have attracted many attentions for their chondrogenic differentiation capacity; however, the roles of EVs in both chondrogenic differentiation of MSCs and cartilage regeneration are poorly understood yet. In the current study, to investigate the differentiation effects of human articular chondrocyte EVs on adipose-derived MSCs, they were cultured in injectable chitosan-hyaluronic acid (CS-HA) hydrogel and then treated with chondrocyte EVs for 21 days. The continuous treatment of EVs performed on MSCs increased chondrogenic genes' expressions ofSOX9andCOL2A1and induced expression of Col II protein. In addition, glycosaminoglycans secretion was detected in the EV-treated MSCs after about 14 days. The therapeutic efficiency of this hydrogel and EVs was studied in a rabbit osteochondral defect model. MRI results revealed that the cartilage regeneration capacity of EV-treated MSCs with CS-HA hydrogel was greater than the untreated MSCs or the EV-treated MSCs without hydrogel. Moreover, histological results showed hyaline-like cartilage in the CS-HA/MSC and CS-HA/EV/MSC groups in the cartilage defect sites. These findings suggested that the chondrocyte-EVs and CS-HA hydrogel could provide the preferable niche for chondrogenic differentiation of MSCs and cartilage regeneration in osteoarthritis cartilage injuries.

Cartilage tissue engineering using injectable functionalized Demineralized Bone Matrix scaffold with glucosamine in PVA carrier, cultured in microbioreactor prior to study in rabbit model.

Using 3D model of injectable scaffolds for cartilage tissue engineering is one of the challenges that should be addressed to avoid invasive surgery for treatment. For this purpose, chondrocytes on Demineralized Bone Matrix (DBM) scaffolds functionalized with glucosamine in 20% polyvinyl alcohol (PVA) as a carrier was applied to the micro-bioreactor in-vitro, then the study was continued on in-vivo stage. Scaffold biocompatibility tests were performed and the mechanical and physicochemical properties were studied showing the fact that DBM was functionalized by Glucosamine, scaffold degradation rate was 53% after 720 h and swelling ratio was 2.5 times after 16 h, injectable scaffold demonstrated better mechanical characteristics (P < 0.05) than other concentrations of PVA. Consequently, in-vitro tests, including live-dead imaging resulting in 99% viability after 14 days (P < 0.001), DAPI staining and scanning electron microscope imaging were performed to determine the number and viability of the cells on the scaffold, showing a cells proliferation property of this group compared with the control after 14 days (P < 0.0001), then relative gene expression was evaluated and protein expression was assessed. The overall chondrogenic gene expression improved (P < 0.05) compared to the control (2D culture). Subsequently, the scaffold were loaded with chondrocytes and injected into the cartilage lesion part After 24 weeks of surgery, MRI and immunocytochemistry were performed. Then all outputs proved that the scaffold plus cell group had a significantly higher topological score (P < 0.0001) than other groups compared to normal cartilage. Finally, studies have shown that transplantation of chondrocytes in DBM, polyvinyl alcohol and glucosamine scaffold through one surgical stage improves cartilage lesion and it can be considered as a breakthrough in tissue engineering.