κ‑Casein-Based Hierarchical Suprastructures and Their Use for Selective Temporal and Spatial Control over Neuronal Differentiation

Functions are diversified by producing hierarchical structures from a single raw material. Biologically compatible milk protein of κ-casein has been employed to fabricate higher-order suprastructures. In the presence of dithiothreitol and heat treatment, κ-casein transforms into amyloid fibrils with distinctive morphology attributable to mechanism-based fibrillar polymorphism. As the fibrils elongate to yield high aspect ratio during high-temperature incubation, the resulting fibrils laterally associate into the liquid crystalline state by forming a two-dimensional fibrillar array. Following a desalting process, the fibrillar arrays turn into a three-dimensional matrix of hydrogel that could be selectively disintegrated by subsequent salt treatment. The hydrogel was demonstrated to be a matrix capable of exhibiting controlled release of bioactive substances like retinoic acid, which led to temporal and spatial control over the differentiation of neuronal cells. Therefore, the hierarchical suprastructure formation derived from the single protein of κ-casein producing one-dimensional protein nanofibrils, a two-dimensional liquid crystalline state and a three-dimensional hydrogel could be widely appreciated in various areas of nanobiotechnology including drug delivery and tissue engineering