Development of Scaffolds with Adjusted Stiffness for Mimicking Disease-Related Alterations of Liver Rigidity

Drug metabolism in patients with liver disease can differ substantially from healthy subjects, and clinical evidence links these changes to increased liver stiffness in fibrosis/cirrhosis. This study developed scaffold-based 3D liver culture models that mimic the stiffness of healthy versus fibrotic/cirrhotic liver to better capture stiffness-dependent changes in hepatic cell behavior relevant to drug testing. The authors fabricated porous pHEMA/BAA cryogel scaffolds with gelatin/collagen additives and screened compositions for pore architecture, permeability, swelling/water uptake, and stiffness. Two scaffold prototypes were selected to approximate healthy-liver rigidity (≈2.9 ± 1.3 kPa) and fibrotic/cirrhotic-like rigidity (stiffer prototypes), then used to culture HepG2 cells. Scaffold pre-incubation in FCS-containing medium strongly improved initial cell attachment, especially on the softer “healthy liver” scaffold, and SEM suggested surface deposits/agglomerates after pre-incubation contributed to improved adherence. Functionally, stiffness altered HepG2 metabolic enzyme activities: cells on softer scaffolds showed higher activity for key enzymes (including CYP3A4, CYP2C9, and UGT), while cells on the stiffer scaffold showed relatively higher CYP1A2 (and a trend toward higher GST). The work concludes that stiffness-tuned 3D scaffolds can model disease-related liver rigidity effects on drug metabolism and may improve prediction of drug behavior in fibrotic/cirrhotic livers compared with conventional 2D plastic culture.