Pathway-Dependent Preferential Selection and Amplification of Variable Self-Assembled Peptide Nanostructures and Their Biological Activities.

We demonstrate the formation of diverse peptide nanostructures, which are "out of equilibrium" based on a single dipeptide gelator. These structures represent the differential energy states of the free energy landscape, which are accessed by differential energy inputs provided by variable self-assembly pathways, that is, heat-cool method or ultrasonication. A higher energy input by the heat-cool method created a thermodynamically favored long entangled nanofibrillar network, while twisted ribbonlike structures were prevalent by ultrasonication. Interestingly, the nanofibrillar network representing the global thermodynamic minima could be accessed by simply melting the kinetically trapped structures as indicated by the thermoreversibility studies. The impact on the material strength was remarkable; gels with an order of magnitude difference in mechanical properties could be fabricated by simply modulating the self-assembly pathways. Interestingly, the thermodynamically favored nanofibrous network promoted cellular adhesion and survival, while a significant number of cells fail to adhere on the kinetically trapped twisted ribbons. Thus, nonequilibrium nanostructures open up new directions to develop advanced functional materials with diverse functions.