Decellularized liver matrix-modified chitosan fibrous scaffold as a substrate for C3A hepatocyte culture

Abstract

A bioreactor filled with functional hepatocytes is a crucial portion of the bio-artificial liver device. However, it is a difficult task to maintain sufficient cell quantity and active hepatocellular function. In this work, we developed a promising scaffold for hepatocyte culture by coating porcine liver extracellular matrix (ECM) on chitosan (CTS) fabrics. Porcine Liver was decellularized using 1% Triton X-100. Solubilized liver ECM was immobilized on CTS fibers surface through cross linking of ECM and CTS with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-Hydroxysuccinimide (NHS). Then the scaffold was characterized by Fourier transformed infrared spectroscopy in attenuated total reflection mode (ATR–FTIR), X-photoelectron spectroscopy (XPS) and water contact angle measurement. The efficacy of modified scaffolds to maintain C3A hepatocytes adhesion, proliferation, bioactivity and functionality in vitro was detected. FTIR spectra and XPS demonstrated the presence of ECM coating on CTS fabric surface. Covalently attached coating significantly improved the binding efficiency between ECM and CTS fabrics, in comparison to the coating by physical absorption. Furthermore, C3A hepatocytes cultured on coated scaffolds showed enhanced cell bioactivity and liver-specific function, such as albumin secretion and urea synthesis, compared with those cultured on untreated scaffolds(p < 0.05). As a promising hepatocyte culture carrier, the ECM coated CTS fabrics could be applied in the biological artificial liver reactor.