Proteins combination on PHBV microsphere scaffold to regulate Hep3B cells activity and functionality: A model of liver tissue engineering system

Abstract The synergistic effects of extracellular matrix (ECM) protein combinations on Hep3B cell proliferation and functions are studied herein. Poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (PHBV) microspheres were covalently conjugated with three types of proteins, collagen (type I), laminin, and fibronectin, using 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide and N‐hydroxysuccinimide cross linkers. Successful conjugations of protein molecules were verified by the presence of nitrogen peaks in X‐ray photoelectron spectroscopy. The densities of grafted proteins were quantified using Micro‐BCA kit. A human hepatoma cell line, Hep3B, was then cultured in vitro on the ECM proteins‐modified microspheres for 2 weeks. Cell proliferation was estimated using MTT method, and two hepatic functions, albumin secretion and P‐450 activity, were evaluated using ELISA and EROD assays, respectively. The results indicated that combination of the three ECM proteins on microsphere surfaces has a significant effect on the proliferation of Hep3B cells, thus better mimicking the in vivo environment for liver tissue engineering. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res 2007