Tendon cell contraction of collagen–GAG matrices in vitro: effect of cross-linking

The contraction of connective tissue cells can play important roles in wound healing and pathological contractures. The effects of this contractile behavior on cell-seeded constructs for tissue engineering have not yet been investigated. The goal of this work was to investigate in vitro tendon cell-mediated contraction of collagen–glycosaminoglycan (GAG) matrices cross-linked using selected methods. Highly porous collagen–GAG sponges were seeded with calf tendon cells and the projected area and DNA content of the sponges measured at 3, 7, 14, and 21 days post-seeding. Immunohistochemistry was performed to determine if α-smooth muscle actin (SMA) was associated with the cell contraction of the matrices. Dehydrothermal (DHT) treatment alone was not sufficient to resist contraction by the seeded tendon fibroblasts. Cross-linking of the collagen–GAG sponges to the extent that the modulus was three times that of sponges treated by DHT alone was necessary to resist contraction. SMA was seen in the cytoplasm of most cells in all sponges at all time periods. The results provide a rational basis for the determination of the mechanical properties of collagen matrices required for engineering certain connective tissues.