Effect of ascorbic acid on protein synthesis and collagen hydroxylation in continuous flow organ cultures of adult mouse periodontal tissues

A continuous flow organ culture system (CFCS) was used to determine the effect of ascorbic acid on the synthesis of collagen and noncollagenous protein by bone of the alveolar process and periodontal ligament in organ cultures of adult mouse periodontium. For the last 24 h of 2 day cultures, 5 microCi/ml 3H-proline was added to the medium. Highly purified collagenase was used to separate the collagenous and noncollagenous proteins and the incorporation of isotope into each fraction measured. Collage synthesized in the presence of less than 10 micrograms/ml ascorbic acid was found to be highly under-hydroxylated (pro:hypro ap. acts. 2.3-3.1) in both tissues. When the ascorbic acid levels were between 25 and 100 micrograms/ml, the synthesis of collagenous proteins was selectively stimulated and hydroxylation significantly improved (pro:hypro sp. acts 1.72-1.89). The effect of ascorbic acid was not related to tissue viability since tissues cultured initially in the absence of ascorbic acid were able to recover completely when compared to controls given ascorbic acid continuously. The proportion of radioactivity in collagen and noncollagenous protein, collagen hydroxylation, and percentage of collagen synthesized as type III (av.23%) in bone of the alveolar process was similar to that found in vivo. However, in the periodontal ligament in vitro the proportion of noncollagenous protein synthesized was increased from 70% to 87% and the percentage of type III collagen increased from 14% to 26% compared to in vivo results.