Hyaluronic acid-dependent change in the extracellular matrix of mouse dermal fibroblasts that is conducive to cell proliferation

ABSTRACT Fluorescein isothiocyanate (FITC)-labelled hyaluronic acid, when incubated with subconfluent cultures of mouse dermal fibroblasts, was incorporated into the extracellular matrix. Deposition of the fluorescent label reached the maximum about 48 h after its addition. Hyaluronic acid decasaccharide, but not octasaccharide, inhibited the incorporation of the fluorescent label, suggesting that at least 10 sugar units in length are necessary for the incorporation of hyaluronic acid. A 2M-urea extract of the cell layer had the ability to bind [3H]hyaluronic acid. Again, the binding was inhibited by hyaluronic acid decasaccharide but not by octasaccharide, suggesting the presence in the urea extract of a hyaluronic acid-binding molecule that may participate in the incorporation of hyaluronic acid. A supramolecular aggregate prepared by rate-zonal sedimentation from the 2M-urea extract contained chondroitin sulphate proteoglycans capable of interacting with hyaluronic acid. Their core molecules were identical in size with those from a hyaluronic acid-binding chondroitin sulphate proteoglycan (PG-M) previously described in chick embryo fibroblasts. Immunofluorescence analyses with anti-proteoheparan sulphate antibodies indicated that both exogenous addition of hyaluronic acid and enhanced synthesis of hyaluronic acid caused a preferential decline in the proteoheparan sulphate level in the extracellular matrix. Subsequent to this change, the cells began transient DNA synthesis. We suggest that hyaluronic aciddependent modulation of the level of proteoheparan sulphate in the extracellular matrix could be a causal event of cell proliferation.