Design and application of a test system for viscoelastic characterization of collagen gels

Characterization and control of the mechanical properties of the extracellular matrix are critical to the interpretation of results of in vitro studies of cultured tissues and cells and for the design of functional engineered constructs. In this work a viscoelastic tensile test system and custom culture chambers were developed and characterized. The system allowed quantification of strain as well as the stresses developed during cyclic viscoelastic material testing. Finite element analysis of the culture chambers indicated that the tensile strains near the actuated ends of the gel were greater than the strains experienced by material in the center of the culture chambers. However, the strain was uniformly distributed over the central substance of the gel, validating the assumption that a homogeneous strain state existed in the central region of the chamber. Viscoelastic testing was performed on collagen gels that were created with three different collagen concentrations. Results demonstrated that there was a significant increase in the dynamic stiffness of the gels with increasing equilibrium strain, collagen concentration, and frequency of applied strain. With increasing strain rate, the phase angle, representing the energy dissipated, dropped initially and then increased at higher rates. Mechanical testing of gels at different time intervals up to 7 days after polymerization demonstrated that the material properties remained stable when appropriate environmental conditions were maintained. The ability to characterize the viscoelastic properties of gels after different periods of culture will allow the quantification of alterations in gel material properties due to changes in cell cytoskeletal organization, cell-matrix interactions, and cellular activity on the matrix. Further, the test device provides a means to apply controlled mechanical loading to growing gel cultures. Finally, the results of this study will provide guidance to the design of further experiments on this substrate.