3D bioprinting of complex tissue scaffolds with in situ homogeneously mixed alginate-chitosan-kaolin bioink using advanced portable biopen.

Control of in situ 3D bioprinting of hydrogel without toxic crosslinker is ideal for tissue regeneration by reinforcing and homogeneously distributing biocompatible reinforcing agent during fabrication of large area and complex tissue engineering scaffolds. In this study, homogeneous mixing, and simultaneous 3D bioprinting of a multicomponent bioink based on alginate (AL)-chitosan (CH), and kaolin was obtained by an advanced pen-type extruder to ensure structural and biological homogeneity during the large area tissue reconstruction. The static, dynamic and cyclic mechanical properties as well as in situ self-standing printability significantly improved with the kaolin concentration for AL-CH bioink-printed samples due to polymer-kaolin nanoclay hydrogen bonding and cross-linking with less amount of calcium ions. The Biowork pen ensures better mixing effectiveness for the kaolin-dispersed AL-CH hydrogels (evident from computational fluid dynamics study, aluminosilicate nanoclay mapping and 3D printing of complex multilayered structures) than the conventional mixing process. Two different cell lines (osteoblast and fibroblast) introduced during large area multilayered 3D bioprinting have confirmed the suitability of such multicomponent bioinks for in vitro even tissue regeneration. The effect of kaolin to promote uniform growth and proliferation of the cells throughout the bioprinted gel matrix is more significant for this advanced pen-type extruder processed samples.