Incorporation of Cross-Linked Gelatin Microparticles To Enhance Cell Attachment and Chondrogenesis in Carboxylated Agarose Bioinks for Cartilage Engineering

Due to the limited regenerative capacity of injured cartilage, surgical intervention using engineered cellular constructs or autologous cell implantation is the best accredited approach to prevent further degeneration and promote a regenerative microenvironment. Advancements in additive manufacturing present opportunities for graft customization through enhanced scaffold design. In bioprinting, an additive manufacturing process, the "bioink" serves as the medium to carry cells but also as a scaffold by imparting form and mechanical attributes to the printed object. In this study, the impact of cross-linked gelatin microparticles (GMPs) on rheological properties and printability of carboxylated agarose (CA) bioink as well as matrix deposition by human nasal chondrocytes (hNCs) was investigated. The introduction of GMPs yielded stiffer bioink formulations, with lower sol-gel transitions that retained the exceptional printability of CA. GMPs served as foci for the attachment of hNCs, improving cellular distribution and bridging the deposited extracellular matrix. After 4 weeks in chondrogenic culture, GMPs containing printed constructs showed enhanced toughness approaching that of the lower end of the spectrum of native cartilage tissue. The incorporation of proteinaceous microparticles might serve as a general concept to promote cellular function in polysaccharide-based bioinks and opens another avenue for engineering 3D-bioprinted microenvironments.