Three-Dimensional Cartilage CultureModel – Cell Culture and Analysis ofChondrogenesis

Abstract: Background: Articular cartilage injuries are a major challenge in orthopedic surgery. Articular cartilage has a low metabolic activity, and it is avascular. Therefore, when injuries occur cartilage has a limited capacity to fully regenerate. Considering that there is a lack of curative surgical approaches to treat cartilage defects, new and more effective interventions are needed. Scaffold-based techniques have a promising potential to address this issue. However, there are still knowledge gaps when it comes to optimal conditions and combination of materials to achieve desired results. Objective: To determine if polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyethylene glycol (PEG) and chitosan (CS)-based scaffolds can be used as matrix to generate cartilage in vitro. Methods: Chondrocytes were cultured for six weeks in different scaffolds composed of PVA, PVP, PEG and CS in different ratios. Sections from the scaffolds and cell pellets were stained with different histological stains, 4′,6-diamidino-2-phenylindole (DAPI) and immunohistochemistry to identify cell nuclei, collagen, proteoglycans, and type II procollagen after the cultivation period. The sections were then examined manually under microscope and photographed. Results: Cells were found in all scaffolds seeded with chondrocytes after the cultivation period. The histological stains had low specificity in identifying cartilage components in scaffolds. The cells cultured with chondrogenic growth factors had higher cell proliferation and more production of collagen and proteoglycans compared to the cells cultured without chondrogenic growth factors. Conclusion: This study indicates that PVA, PVP, PEG and CS polymers can be used to culture chondrocytes in vitro. Chondrogenic growth factors seem to have an important role in generating cartilage in vitro.