Combination of proteins on PHBV microsphere scaffold to regulate Hep3B cells activity and functionality for an in vitro model of liver tissue engineering

Abstract

The synergistic effects of extracellular matrix (ECM) proteins combination 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 by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide cross linkers. The densities of grafted proteins were quantified by using a Micro-BCA kit. A human hepatoma cell line, Hep3B, was then cultured in vitro on the ECM proteins modified microspheres for two 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.