Local mechanical stiffness influences cell behavior, and thus cell culture scaffolds should approximate the stiffness of the tissue type from which the cells are derived. In synthetic hydrogels, this has been difficult to achieve for very soft tissues such as neural. This work presents a method for reducing the stiffness of mechanically and biochemically tunable synthetic poly(ethylene glycol) diacrylate hydrogels to within the soft tissue stiffness regime by altering the organization of the crosslinking sites. A soluble allyl-presenting monomer, which has a higher propensity for chain termination than acrylate monomers, was introduced into the PEG-diacrylate hydrogel precursor solution before crosslinking, resulting in acrylate-allyl competition and a reduction in gel compressive modulus from 5.1 ± 0.48 kPa to 0.32 ± 0.09 kPa. Both allyl monomer concentration and chemical structure were shown to influence the effectiveness of competition and change in stiffness. Fibroblast cells demonstrated a 37% reduction in average cell spread area on the softest hydrogels produced as compared to cells on control hydrogels, while the average percentage of neural cells extending neurites increased by 41% on these hydrogels, demonstrating the potential for this technology to serve as a soft tissue culture system.
Keywords: PEGDA; cell culture scaffold; hydrogel; mechanotransduction; substrate stiffness; tissue engineering.