Self-Healing Scaffolding Technology with Strong, Reversible Interactions under Physiological Conditions for Engineering Marbled Cultured Meat

Abstract

Cultured meat offers a sustainable alternative to animal farming, with the potential to reduce environmental impacts and improve food security. However, recapitulating natural meat marbling remains a significant challenge. This study presents a straightforward technology for achieving precise marbling patterns in large-scale cultured meat using self-healing hydrogels containing boronic acid-conjugated chitosan. Unlike conventional hydrogels, which require nonphysiological conditions for strong, reversible bonding, our system achieves robust reversible bonding at neutral pH through a unique mechanism: the nucleophilic groups of chitosan facilitate boronic acid-diol bond formation, exhibiting half the strength of a typical covalent bond, as demonstrated by nanomechanics analysis. The hydrogels form dual reversible networks of boronic acid-diol and hydrogen bonds, enabling self-healing and tunable stiffness. Biocompatibility studies confirm that they support the growth of mouse-derived cells and bovine-derived primary muscle cells. Each hydrogel variant optimizes mechanotransduction for the distinct requirements of fat or muscle cell culture and differentiation. This self-healing scaffolding technology enables the seamless assembly of muscle and fat monocultures into centimeter-thick meat with micrometer-scale marbling patterns, tailoring organoleptic properties and nutritional profiles without the need for meat glues or processing equipment.