Regeneration of meniscus tissue using adipose mesenchymal stem cells-chondrocytes co-culture on a hybrid scaffold: In vivo study.

The meniscus has poor intrinsic regenerative capacity and its damage inevitably leads to articular cartilage degeneration. We focused on evaluating the effects of Polyvinyl alcohol/Chitosan (PVA/Ch) scaffold seeded by adipose-derived mesenchymal stem cell (ASC) and articular chondrocytes (AC) in meniscus regeneration. The PVA/Ch scaffolds with different molar contents of Ch (Ch1, Ch2, Ch4 and Ch8) were cross-linked by pre-polyurethane chains. By increasing amount of Ch tensile modulus was increased from 83.51 MPa for Ch1 to 110 MPa for Ch8 while toughness showed decrease from 0.33 mJ/mm in Ch1 to 0.11 mJ/mm in Ch8 constructs. Moreover, swelling ratio and degradation rate increased with an increase in Ch amount. Scanning electron microscopy imaging was performed for pore size measurement and cell attachment. At day 21, Ch4 construct seeded by AC showed the highest expression with 24.3 and 22.64 folds increase in collagen II and aggrecan (p ≤ 0.05), respectively. Since, the mechanical properties, water uptake and degradation rate of Ch4 and Ch8 compositions had no statistically significant differences, Ch4 was selected for in vivo study. New Zealand rabbits were underwent unilateral total medial meniscectomy and AC/scaffold, ASC/scaffold, AC-ASC (co-culture)/scaffold and cell-free scaffold were engrafted. At 7 months post-implantation, macroscopic, histologic, and immunofluorescent studies for regenerated meniscus revealed better results in AC/scaffold group followed by AC-ASC/scaffold and ASC/scaffold groups. In the cell-free scaffold group, there was no obvious meniscus regeneration. Articular cartilages were best preserved in AC/scaffold group. The best histological score was observed in AC/scaffold group. Our results support that Ch4 scaffold seeded by AC alone can successfully regenerate meniscus in tearing injury and ASC has no significant contribution in the healing process.