Publications

Anisotropy is an important cue for neural organization during morphogenesis and healing, contributing to the mechanical and functional properties of neural tissues. The ability to replicate such anisotropy in vitro holds great promise for the development of effective regeneration strategies. In this work, interfacial polyelectrolyte complexation (IPC) is applied to fabricate microfibers from charged ECM components without any chemical modification. Using flow-focusing microfluidics, collagen (Col) and glycosaminoglycans (GAGs), such as chondroitin sulfate (CS) or heparin (Hep), form Col/CS and Col/Hep interfacial complexes that coalesce as IPC microfibers. These fibers are flexible and absorb large amounts of water but remain stable under physiological conditions. At these conditions, the tensile strength of the assembled Col/GAG microfibers is similar to the strength of the neural tissue. The fibers are biocompatible and biofunctional; PC12 neural cells adhere and orient longitudinally to the fibers. Moreover, Col/CS microfibers promote the formation of neural processes. The results demonstrate that the microfluidic-assisted IPC complexation enables the assembly of ECM mimics by synergetic integration of anisotropic, chemical, and mechanical cues that boost the development of neural cells.

JTD Keywords: Cells, Chondroitin sulfate, Collagen, Fibers, Glycosaminoglycans, Heparin, Microfibers

The histopathological hallmarks of Alzheimer disease are the self-aggregation of the amyloid beta peptide (A beta) in extracellular amyloid fibrils and the formation of intraneuronal Tau filaments, but a convincing mechanism connecting both processes has yet to be provided. Here we show that the endogenous polysaccharide chondroitin sulfate B (CSB) promotes the formation of fibrillar structures of the 42-residue fragment, A beta(1-42). Atomic force microscopy visualization, thioflavin T fluorescence, CD measurements, and cell viability assays indicate that CSB-induced fibrils are highly stable entities with abundant beta-sheet structure that have little toxicity for neuroblastoma cells. We propose a wedged cylinder model for A beta(1-42) fibrils that is consistent with the majority of available data, it is an energetically favorable assembly that minimizes the exposure of hydrophobic areas, and it explains why fibrils do not grow in thickness. Fluorescence measurements of the effect of different A beta(1-42) species on Ca2+ homeostasis show that weakly structured nodular fibrils, but not CSB-induced smooth fibrils, trigger a rise in cytosolic Ca2+ that depends on the presence of both extracellular and intracellular stocks. In vitro assays indicate that such transient, local Ca2+ increases can have a direct effect in promoting the formation of Tau filaments similar to those isolated from Alzheimer disease brains.

JTD Keywords: AFM, Alzheimers-disease, Chondroitin sulfate, Heparan-sulfate, Lipid-bilayers, Beta-peptide, In-vitro, Neurodegenerative diseases, Extracellular-matrix, Prion protein