Expression of hyaluronan synthases and hyaluronidases in the MG63 osteoblast cell line

Abstract

The expression of hyaluronan synthases (1, 2 and 3) and hyaluronidases (1, 2, 3, 4 and PH20) was examined in the MG63 osteoblast cell line induced to mineralize in vitro and compared to the rate of glycosaminoglycan production. Real-time PCR analysis demonstrated a 13-fold decrease in hyaluronan synthase 3 expression in mineralising MG63 cells; no significant change in hyaluronan synthase 2 expression in mineralising cells and hyaluronah synthase 1 was not expressed. In mineralising MG63 cells a 62-fold increase in hyaluronidase 2, a 13-fold increase in hyaluronidase 3, and a 3-fold increase in hyaluronidase 4 expression were observed when compared to non-mineralising cells; hyaluronidase 1 and PH20 expression was not detected. After 5 weeks in mineralising culture conditions a 2-fold increase in total 3H-glucosamine incorporation was observed in cells when compared to 24 h or 5 week control cultures. This was made up of a 5-fold decrease in hyaluronan production, a 2-fold increase in chondroitin sulphate/dermatan sulphate and a 10-fold increase in 3H-glucosamine incorporation into the non-glycosaminoglycan fraction. A 3-fold increase in 35SO4 incorporation into chondroitin sulphate/dermatan sulphate was also observed. Thus there is co-ordinate expression of genes that control hyaluronan metabolism such that there is a general decrease in the expression of hyaluronan synthases, an increase in the expression of hyaluronidases and a corresponding decrease in hyaluronan production by mineralising MG63 cells.