Glycosaminoglycan functionalization of mechanically and topologically defined collagen i matrices

Abstract

Collagen I and glycosaminoglycans (GAGs) are major components of the extracellular matrix in mammals and widely used for in vitro cell culture matrices. While composition, network microstructure and mechanics of these matrices sensitively determine cell fate, they are hard to adjust independently during matrix reconstitution. We report on a sequential preparation procedure of collagen I matrices, which allows a defined adjustment of network topology and mechanics in combination with GAG functionalization. Collagen I solution concentrations of 1.5 to 7 mg ml-1 allowed to vary topology (pore size) and elasticity of resulting networks with Young's moduli of 5 to 220 Pa. Zero-length crosslinking using carbodiimide chemistry increased Young's modulus 3 to 5 times without changing network topology. An optional covalent binding of hyaluronan and synthetically sulfated hyaluronan to the preformed matrices led to topologically unaffected networks with a stable functionalization with ∼30 μg GAG per mg collagen. While sulfated GAGs were stably attached to collagen I networks via physisorption or covalent binding at neutral and acidic conditions, non-sulfated hyaluronan required acidic conditions and covalent binding for stable attachment. In conclusion, this approach provides options to independently adjust biophysical and biochemical parameters of collagen I networks for in vitro studies.