Design of Hyaluronic Acid Hydrogels to Promote Neurite Outgrowth in Three Dimensions.
A hyaluronic acid (HA)-based extracellular matrix (ECM) platform with independently tunable stiffness and density of cell-adhesive peptide (RGD, arginine-glycine-aspartic acid) that mimics key biochemical and mechanical features of brain matrix has been designed. We demonstrated here its utility in elucidating ECM regulation of neural progenitor cell behavior and neurite outgrowth. The analysis of neurite outgrowth in 3-D by two-photon microscopy showed several important results in the development of these hydrogels. First, the ability of neurites to extend deeply into these soft HA-based matrices even in the absence of cell-adhesive ligand further confirms the potential of HA hydrogels for central nervous system (CNS) regeneration. Second, the behavior of hippocampal neural progenitor cells differed markedly between the hydrogels with a storage modulus of 400 Pa and those with a modulus of 800 Pa. We observed an increased outgrowth and density of neurites in the softest hydrogels (G' = 400 Pa). Interestingly, cells seeded on the surface of the hydrogels functionalized with the RGD ligand experienced an optimum in neurite outgrowth as a function of ligand density. Surprinsingly, neurites preferentially progressed inside the gels in a vertical direction, suggesting that outgrowth is directed by the hydrogel structure. This work may provide design principles for the development of hydrogels to facilitate neuronal regeneration in the adult brain.
Grenoble Alpes University , Centre de Recherches sur les Macromolécules Végétales (CERMAV-CNRS), 601, rue de la Chimie, BP 53, Grenoble 38041 Cedex 9, France.
Dominte
D
Tarus
Platform Intravital Microscopy, France Life Imaging, Grenoble Alpes University , INSERM U1205, 17 rue des Martyrs, 38054 Grenoble, France.
Lauriane
L
Hamard
Platform Intravital Microscopy, France Life Imaging, Grenoble Alpes University , INSERM U1205, 17 rue des Martyrs, 38054 Grenoble, France.
Flavien
F
Caraguel
Grenoble Alpes University , INSERM U1205, 17 rue des Martyrs, 38054 Grenoble, France.
Didier
D
Wion
Grenoble Alpes University , Centre de Recherches sur les Macromolécules Végétales (CERMAV-CNRS), 601, rue de la Chimie, BP 53, Grenoble 38041 Cedex 9, France.
Anna
A
Szarpak-Jankowska
Platform Intravital Microscopy, France Life Imaging, Grenoble Alpes University , INSERM U1205, 17 rue des Martyrs, 38054 Grenoble, France.
Boudewijn
B
van der Sanden
Grenoble Alpes University , Centre de Recherches sur les Macromolécules Végétales (CERMAV-CNRS), 601, rue de la Chimie, BP 53, Grenoble 38041 Cedex 9, France.
Rachel
R
Auzély-Velty
D005109:Extracellular Matrix; D006820:Hyaluronic Acid / Q000737:chemistry*; D020100:Hydrogels; D016501:Neurites; D000071446:Neuronal Outgrowth
D020100:Hydrogels; D006820:Hyaluronic Acid
hyaluronic acid; hydrogel; neural progenitor cells; neurite outgrowth; two-photon microscopy