Nanostructuring PEG-fibrinogen hydrogels to control cellular morphogenesis

The nanostructuring of hydrogel scaffolds used in tissue engineering aims to provide an ability to control cellular morphogenesis through defined cell–matrix interactions. Toward this objective, we developed a method that alters the molecular network structure of biosynthetic hydrogel scaffolds made from crosslinked poly(ethylene glycol)-fibrinogen conjugates (PEG-fibrinogen, PF). The modifications were based on Pluronic® F127 micelles that were formed in the hydrogel precursor solution and that altered the hydrogel network assembly during photopolymerization crosslinking. Two variations of the cell-encapsulating hydrogels (high and low crosslinking density) were prepared with three concentrations of Pluronic® F127 (3%, 7%, 10% w/v). Quantitative morphometrics were used to characterize fibroblast shape parameters (both transient and stable) in all hydrogels, and rheological characterizations were used to measure the elastic (storage) component of the complex shear modulus of these hydrogels. The morphometric data was then correlated to both the nanostructure and modulus of the hydrogels for day 1 and day 4 in culture. These correlations revealed that structural features imparted by the Pluronic® F127 micelles were able to reverse the normally strong correlations found between indicators of cell spreading and the hydrogel’s mechanical properties. Therefore, the data supports the conclusion that nanostructural features in the encapsulating hydrogel culture environment can facilitate better cell spreading in a dense hydrogel milieu, simply by introducing imperfections into the network structure. This research also provides further prospective regarding biocompatible approaches toward making structural modifications to hydrogel scaffolds for the purpose of 3-D cell culture and tissue engineering.