De Novo Design of Functional Co-Assembling Organic-Inorganic Hydrogels for Hierarchical Mineralization and Neovascularization

Abstract

Synthetic nanostructured materials incorporating both organic and inorganic components offer a unique, powerful and versatile class of materials for widespread applications due to the distinct, yet complementary, nature of the intrinsic properties of the different constituents. We report a supramolecular system based on synthetic nanoclay (Laponite™, Lap) and peptide amphiphiles (PAs, PAH3) rationally designed to co-assemble into nanostructured hydrogels with high structural integrity and a spectrum of bioactivities. Spectroscopic and scattering techniques and molecular dynamic simulation approaches were harnessedto confirm that PAH3 nanofibers electrostatically adsorbed and conformed to the surface of Lapnanodisks. Electron and atomic force microscopies also confirmed an increase in diameter and surface areaof PAH3nanofibers after co-assembly with Lap. Dynamic oscillatory rheology revealed that the co-assembled PAH3-Laphydrogels displayed high stiffness and robust self-healing behaviour while gas adsorption analysis confirmed a hierarchical and heterogeneous porosity. Furthermore, this distinctive structure within the three-dimensional matrix(3D) provided spatial confinement for the nucleation and hierarchical organization of high-aspect ratio hydroxyapatite nanorods into well-defined spherical clusters within the 3Dmatrix. Applicability of the organic-inorganic PAH3-Laphydrogels was assessed in vitrousing human bone marrow-derived stromal cells(hBMSCs) and ex vivousing a chick chorioallantoic membrane (CAM) assay. The results demonstrated that the organic-inorganic PAH3-Laphydrogels promote human skeletal cell proliferation and, upon mineralization, integrate with the CAM, are infiltrated by blood vessels, stimulate extracellular matrix production, and facilitate extensive mineral deposition relative to the controls