Ascorbate modulation of bovine chondrocyte growth, matrix protein gene expression and synthesis in three-dimensional collagen sponges.

Marie-Claire Ronzière (1) , Stéphane Roche (1) , Jérome Gouttenoire (1) , Olivier Demarteau (1) , Daniel Herbage (1) , Anne-Marie Freyria (1)

1 IBCP - Institut de biologie et chimie des protéines [Lyon]

This report completes a previous study on the growth and metabolism of fetal bovine epiphyseal chondrocytes cultured, within native or cross-linked collagen sponges carried out without the addition of fresh ascorbate. At low initial cell density (2.3 x 10(6)cells/cm(3)) cell proliferation and a low matrix deposition were observed, whereas at high initial cell density (2.3 x 10(7)cells/cm(3)) there was an absence of cell proliferation, but the deposition of a cartilage-like matrix was measured. In both cases, only traces of type I collagen (marker of chondrocyte dedifferentiation) were detected. In this report, we observed, after 1 month in culture with ascorbate, in both type of scaffolds and initial cell densities, an increase in cell proliferation (2-fold) and in expression of genes encoding for collagen types I, II, X and MMP-2 and -13, but no change in the level of matrix deposition (collagen and GAG). With regard to the proteins present, the main differences with or without ascorbate concerned the increase of neosynthesised type I collagen (up to 35% of the total collagen deposited in the sponge) and of the MMP-2 active form. In conclusion, these results show that ascorbate is an important factor to consider when preparing cartilage constructs for its action on chondrocyte phenotype modulation and proliferation.This report completes a previous study on the growth and metabolism of fetal bovine epiphyseal chondrocytes cultured, within native or cross-linked collagen sponges carried out without the addition of fresh ascorbate. At low initial cell density (2.3 x 10(6)cells/cm(3)) cell proliferation and a low matrix deposition were observed, whereas at high initial cell density (2.3 x 10(7)cells/cm(3)) there was an absence of cell proliferation, but the deposition of a cartilage-like matrix was measured. In both cases, only traces of type I collagen (marker of chondrocyte dedifferentiation) were detected. In this report, we observed, after 1 month in culture with ascorbate, in both type of scaffolds and initial cell densities, an increase in cell proliferation (2-fold) and in expression of genes encoding for collagen types I, II, X and MMP-2 and -13, but no change in the level of matrix deposition (collagen and GAG). With regard to the proteins present, the main differences with or without ascorbate concerned the increase of neosynthesised type I collagen (up to 35% of the total collagen deposited in the sponge) and of the MMP-2 active form. In conclusion, these results show that ascorbate is an important factor to consider when preparing cartilage constructs for its action on chondrocyte phenotype modulation and proliferation.

Mots clés: Ascorbate; Bovine chondrocytes; Collagen biomaterials; Cartilage engineering

Domaines: Biochimie, Biologie Moléculaire

Marie-Claire Ronzière, Stéphane Roche, Jérome Gouttenoire, Olivier Demarteau, Daniel Herbage, et al.. Ascorbate modulation of bovine chondrocyte growth, matrix protein gene expression and synthesis in three-dimensional collagen sponges.. Biomaterials, 2003, 24, pp.851-861. ⟨10.1016/S0142-9612(02)00418-0⟩. ⟨hal-00313309⟩