Bioinspired Bioinks for the Fabrication of Chemomechanically Relevant Standalone Disease Models of Hepatic Steatosis

Hepatotoxicity‐related issues are poorly predicted during preclinical experimentation, as its relevance is limited by the inadequacy to screen all the non‐physiological subclasses of the population. These pitfalls can be solved by implementing complex in vitro models of hepatic physiology and pathologies in the preclinical phase. To produce these platforms, extrusion‐based bioprinting is focused on, since it allows to manufacture tridimensional cell‐laden constructs with controlled geometries, in a high‐throughput manner. Different bioinks, whose formulation is tailored to mimic the chemomechanical environment of hepatic steatosis, the most prevalent hepatic disorder worldwide, are proposed. Internally crosslinked alginate hydrogels are chosen as structural components of the inks. Their viscoelastic properties (G′ = 512–730 Pa and G″ = 94–276 Pa, depending on frequency) are tuned to mimic those of steatotic liver tissue. Porcine hepatic ECM is introduced as a relevant biochemical cue. Sodium oleate is added to recall the accumulation of lipids in the tissue. Downstream analyses on 14‐layered bioprinted structures cultured for 10 days reveal the establishment of steatotic‐like features (intracellular lipid vesicles, viability decrease up to ≈50%) without needing external conditionings. The presented bioinks are thus suitable to fabricate complex models of hepatic steatosis to be implemented in a high‐throughput experimental frame. s‐Hep3Gel is a bioink for extrusion‐based printing that mimics in vitro the hepatic steatosis environment. Alginate hydrogels crosslinked through internal gelation are exploited to mimic the mechanical properties of steatotic liver. Bioinks are supplemented with ECM powder to provide cells with key biochemical cues, and with sodium oleate to mimic the pathological build‐up of fatty acids in the surrounding tissue.