Self-assembled chitosan/gelatin nanofibrous aggregates incorporated thermosensitive nanocomposite bioink for bone tissue engineering

Chitosan-based thermosensitive bioink can be a potential option as bioinks for bone tissue engineering because of their excellent biocompatibility and crosslinker-free gelation at physiological temperature. However, their low mechanical strength, poor printability, and low post-printing cell viability are some of their limitations. In this work, self-assembled nanofibrous aggregates of chitosan and gelatin were prepared and incorporated in chitosan-based bioinks to enhance printability, mechanical properties, post-printing cell viability, and proliferation. Subsequently, the optimal concentration of nanohydroxyapatite was determined, and the potential of the nanocomposite bioink was evaluated. Physiochemical, mechanical, and in vitro characterizations were carried out for the developed nanocomposite bioink. The bioink had optimum printability at 10 % nanohydroxyapatite and cell viability >88 %. The composite bioink had a low water uptake capacity (2.5 %) and degraded within 3 weeks in the presence of lysozyme. Mechanical characterization revealed an elastic modulus of about 15.5 kPa. Rheological analysis indicated a higher storage modulus of the bioink samples at 37 °C. ALP activity of 36.8 units/ml after 14 days of scaffold culture in osteogenic media indicated high cellular activity. These results suggested that the incorporation of osteogenic nanohydroxyapatite and nanofibrous aggregates improved the overall osteogenic and physiochemical potential of the thermosensitive bioink.