Isolation and partial characterization of hyaluronan-protein-collagen complex (HA-PC) from fetal sheep skin of different gestational ages.

Hyaluronic acid (HA) has a positive effect on cell migration, differentiation and wound healing. Earlier work from our laboratory has shown the presence of biologically active proteins associated with HA. The protein associated with HA of fetal sheep skin varies in molecular weight depending on its gestational age. Specifically, the protein profile changes at 125 days of gestation, from a 60 KDa protein to a smaller protein of about 21 KDa. This time period coincides with the time that scarring becomes apparent in fetal sheep skin wounds. In this study, we have quantified changes in the proteins associated with HA with increasing gestational age, obtained amino acid profiles of these proteins with increasing gestational age, and proposed the existence of an HA-associated protein-collagen complex (HA-PC) which may serve as a scaffold for wound healing. Our results indicate that HA-PC content decreases from 42% of the dry weight at 75 days of gestation to 22% at 125 days of gestation. Protein content, in contrast, increases to 40% of the dry weight at 140 days of gestation. At the same time, collagen content increases from < 1% of the dry weight at 75 days to > 10% at 140 days. The increase in collagen content may account for the increase in total protein seen at 140 days. The expression of varying HA-PC's at different gestational ages may influence the kinetics of collagen fibrillogenesis and thus account for the previously noted late gestational change from "scarless" wound healing to "adult-like" wound healing in fetal sheep.