Effects of Fibrinolytic Inhibitors on Chondrogenesis of Bone-marrow Derived Mesenchymal Stem Cells in Fibrin Gels

Abstract

The objective of this study was to examine the effect of two fibrinolytic inhibitors, aprotinin and aminohexanoic acid, on chondrogenesis of rabbit bone marrow mesenchymal stem cells (BM-MSCs). Rabbit BM-MSCs were obtained from the tibias and femurs of New Zealand White rabbits. Cell-fibrin constructs were made by mixing a cell-fibrinogen (10(7) cells/ml; 40 mg/ml fibrinogen) solution with a thrombin (5 IU/ml) solution and then divided into four groups: aprotinin control, aprotinin + transforming growth factor beta (TGF-beta), aminohexanoic acid control, and aminohexanoic acid + TGF-beta. Each of these groups was further treated with three different concentrations of inhibitors and the TGF-beta groups were treated with 10 ng/ml of TGF-beta1. The chondrogenic gene expressions, DNA content, and glycosaminoglycan content of samples were analyzed after 14 days of culture. The aprotinin groups exhibited significantly higher levels of aggrecan gene expression and glycosaminoglycan content than the aminohexanoic acid groups. However, inhibitor neither influenced gene expression of type II collagen nor proliferation (i.e., DNA content) of BM-MSCs. These findings suggest that fibrinolytic inhibitors used to control degradation of fibrin clot may influence TGF-beta-induced chondrogenesis of BM-MSCs.