Abstract
Cell encapsulating scaffolds are necessary for the study of cellular mechanosensing of cultured cells. However, conventional scaffolds used for loading cells in bulk generally fail at low compressive strain, while hydrogels designed for high toughness and strain resistance are generally unsuitable for cell encapsulation. Here we describe an alginate/gelatin methacryloyl interpenetrating network with multiple crosslinking modes that is robust to compressive strains greater than 70%, highly biocompatible, enzymatically degradable and able to effectively transfer strain to encapsulated cells. In future studies, this gel formula may allow researchers to probe cellular mechanosensing in bulk at levels of compressive strain previously difficult to investigate.
Highlights
Mechanical strains and stresses influence cell behavior and outcome in a wide variety of contexts [1,2,3,4,5,6]
To permit detailed analysis of the biologic changes resulting from compression, the ideal material would permit facile and quick recovery of encapsulated cells for further analyses. To develop such a gel, we investigated the use of both interpenetrating networks (IPNs) and multiple crosslinking mechanisms, two techniques known to increase hydrogel toughness [28,29]
We investigated the sol viscosity and Ca2+ ionic gelation kinetics of three different types of alginate: (1) unmodified high molecular weight (HMW) with a molecular weight close to 250 kDa, (2) intermediate molecular weight polymers generated by autoclaving HMW
Summary
Mechanical strains and stresses influence cell behavior and outcome in a wide variety of contexts [1,2,3,4,5,6]. Cells are typically loaded in bulk within tissue, where the loading affects the local osmotic environment and may alter paracrine signaling between cells [8]. For this reason, application of exogenous strain to engineered tissues or cells in hydrogels is often the most physiologically relevant way to study cellular mechanosensing pathways in vitro [7]. The primary methods for studying cell responses to mechanical loading in bulk are the application of loads to tissue explants and to cells seeded in scaffolds. Compression of tissue explants allows cells to be loaded in their native milieu, this approach cannot be used for cells grown in vitro (e.g., cell lines) and limits researchers’
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