Cell stage-dependent effects of ascorbic acid on cultured porcine bone cells

Pig bone cells were isolated from fetuses or young animals. In culture, they proliferated for 6 days, became confluent and began differentiating. The effects of ascorbic acid (AsA) on cell proliferation, alkaline phosphatase (ALP) activity, non-collagenous protein (NCP) and collagen synthesis, were studied by adding AsA to the medium at different times during culture. AsA affected fetal and post-natal cells similarly: ALP activity, NCP and collagen synthesis were markedly reduced in cells treated before confluence, but were strongly and dose-dependently stimulated in cells treated after confluence. AsA also stimulated cell proliferation. The cell stage-dependent action of AsA suggests that it may interfere with differentiation. The effects of AsA on ALP activity and DNA content were not coupled to its effect on collagen synthesis, raising the question of whether AsA action is matrix-mediated.