Ascorbate stimulates type I and type III collagen in human Tenon's fibroblasts

Purpose: To better understand wound healing after glaucoma filtration surgery by measuring the production of type I and type III collagen in cultured Tenon's fibroblasts and determine the effect of ascorbic acid on collagen subtype production. Methods: An ELISA-type dot blot assay was used to directly measure the production of types I and III collagen by subconfluent cultures of fibroblasts from human Tenon's capsule. Because ascorbic acid is both high in aqueous humor and necessary for the production of collagen, we measured the dose response of type I and type III collagen production to ascorbic acid. Results: Ascorbic acid stimulated an increase in collagen production that reached a maximum level at 100 μg/ml. This is approximately half of the ascorbic acid concentration found in human aqueous humor. Unlike previous reports, we found no toxic effects from ascorbic acid at concentrations as high as 250 yg/ml over a 24-hour period. The lack of toxicity may result from the use of serum-free media in the assay. Conclusions: This culture system will be useful for exploring factors that may alter collagen production and could potentially affect wound healing.