Abstract
Cell-based therapy using chondrocytes for cartilage repair suffers from chondrocyte dedifferentiation. In the present study, the effects of an integrated three-dimensional and dynamic culture on rabbit articular chondrocytes were investigated. Cells (passages 1 and 4) were encapsulated in alginate gel beads and cultured in spinner flasks in chondrogenic and chondrocyte growth media. Subcutaneous implantation of the cell-laden beads was performed to evaluate the ectopic chondrogenesis. It was found that cells remained viable after 35 days in the three-dimensional dynamic culture. Passage 1 cells demonstrated a proliferative growth in both media. Passage 4 cells showed a gradual reduction in DNA content in growth medium, which was attenuated in chondrogenic medium. Deposition of glycosaminoglycans (GAG) was found in all cultures. While passage 1 cells generally produced higher amounts of GAG than passage 4 cells, GAG/DNA became similar on day 35 for both cells in growth media. Interestingly, GAG/DNA in growth medium was greater than that in chondrogenic medium for both cells. Based on GAG quantification and gene expression analysis, encapsulated passage 1 cells cultured in growth medium displayed the best ectopic chondrogenesis. Taken together, the three-dimensional and dynamic culture for chondrocytes holds great potential in cartilage regeneration.
Highlights
IntroductionEstablished clinical treatments for cartilage damages often lead to inferior fibrocartilage-like reparative tissue compared to native hyaline cartilage
Articular cartilage has a very poor self-repair capability
Cells were collected by centrifuging and resuspended in chondrocyte growth medium, which consisted of high-glucose Dulbecco’s modified Eagle’s medium (DMEM), 10% fetal bovine serum (FBS; HyClone), 0.1 mM nonessential amino acids (Invitrogen), 0.4 mM proline (Invitrogen), 0.05 mg/mL vitamin C (Invitrogen), 100 units/mL penicillin (Invitrogen), and 100 units/mL streptomycin (Invitrogen)
Summary
Established clinical treatments for cartilage damages often lead to inferior fibrocartilage-like reparative tissue compared to native hyaline cartilage. Autologous chondrocyte implantation (ACI), a cell-based strategy, holds great promise in regenerating cartilage defects [1]. In this approach, chondrocytes are acquired from non-load-bearing areas of articular cartilage, expanded in vitro and injected into defects as a suspension [2]. Several clinical studies have documented hyaline-like repair tissue and functional improvements with ACI, its long-term efficacy remains controversial [1, 3]. Expansion of primary chondrocytes on two-dimensional (2D) plastic surfaces in vitro inevitably leads to the dedifferentiation [4]. Due to the loss of cartilage phenotype, dedifferentiated chondrocytes might play negative effects in cartilage regeneration [6]
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