Hyaluronic Acid Macromer Molecular Weight Dictates the Biophysical Properties and in Vitro Cellular Response to Semisynthetic Hydrogels.

In situ-forming hydrogels present a promising approach for minimally invasive cell transplantation and tissue regeneration. Among prospective materials, hyaluronic acid (HyA) has displayed great potential, owing to its inherent biocompatibility, biodegradation, and ease of chemical modification. However, current studies in the literature use a broad range of HyA macromer molecular weights (MWs) from <100 kDa to 1 MDa with no consensus regarding an optimal MW for a specific application. We investigated the effects of different HyA macromer MWs on key biophysical properties of semisynthetic hydrogels, such as viscosity, gelation time, shear storage modulus, molecular diffusion, and degradation. Using higher-MW HyA macromers leads to quicker gelation times and stiffer, more stable hydrogels with smaller mesh sizes. Assessment of the potential for HyA hydrogels to support network formation by encapsulated vascular cells derived from human-induced pluripotent stem cells reveals key differences between HyA hydrogels dependent on macromer MW. These effects must be considered holistically to address the multifaceted, nonmonotonic nature of HyA MW on hydrogel behavior. Our study identified an intermediate HyA macromer MW of 500 kDa as providing optimal conditions for a readily injectable, in situ-forming hydrogel with appropriate biophysical properties to promote vascular cell spreading and sustain vascular network formation in vitro.