Fibrin concentration affects ACL fibroblast proliferation and collagen synthesis

Abstract

Fibrin is a frequently used biomaterial in surgery and tissue engineering. While it has been shown that fibrin supports cellular proliferation and biosynthesis, there is a scarcity of studies focusing on the effects of fibrin concentration. The objective of this study is to assess the effect of fibrin concentrations around the physiological concentration of 3mg/ml on the behavior of ligament fibroblasts. Fibroblasts were obtained from the anterior cruciate ligaments of four pigs and seeded throughout fibrin gels of either 1, 3, or 6 mg/ml fibrin. The gels were collected at 2, 6, and 10 days for measurement of DNA and collagen content. We found that both DNA and collagen content increased significantly over time in gels made with all concentrations of fibrin. However, the increases were significantly lower in gels made with the higher concentrations of fibrin (3 and 6 mg/ml). Microscopic assessment of FITC-labeled gels showed a decrease in pore size at high fibrin concentrations, which might be a reason for the observed effect on bioactivity. To enhance cell behavior and thus clinical results fibrin applications should build on physiologic or sub-physiologic concentrations, and those with higher concentrations, such as currently available sealants, should be used cautiously.

Figure 1

shows representative areas of FITC-labeled clots for the three different concentrations. Note the decrease in porosity from low (1 mg/ml and 3 mg/ml) to high (6 mg/ml) concentrations. (100× magnification, size bar represents 100 μm).