Effects of collagen 1, fibronectin, laminin and hyaluronic acid concentration in multi-component gels on neurite extension

Recovery after peripheral nerve injury remains a significant challenge. Extracellular matrix proteins and hydrogels of extracellular matrix components have been shown to improve regeneration in peripheral nerve entubulation models, especially over long distances. The chemical properties, ligand identity and density, and mechanical properties of the hydrogel can affect neurite extension. However, the importance of combinatorial effects between different components in co-gels of several extracellular matrix components is unclear. In this study, we investigated neurite extension from explanted dorsal root ganglia cultured within co-gels made from laminin, fibronectin, collagen 1 and hyaluronic acid. Laminin had a strong, dose-dependent effect on both neurite length and outgrowth. Fibronectin was slightly, but generally not significantly, inhibitory to neurite extension. The concentration of collagen 1 and hyaluronic acid did not have significant effects on neurite extension. The combinatorial effects among the four components were additive rather than synergistic. A co-gel made with 1.5 mg/ml collagen 1 and 1.5 mg/ml laminin was optimum in this study, resulting in an average neurite length of 1532 +/- 91 mu m versus 976 +/- 32 mu m for controls, and an increase in overall volume outgrowth (reflecting neurite length and branching) of 85.9 +/- 9.3% over controls. This co-gel provides a mechanically stable scaffold with high ligand density and biochemical affinity. The results of this study support the use of co-gels of laminin and collagen 1 for promoting regeneration in peripheral nerve injuries and suggest that interactions among hydrogel components are not significant.