Proteins combination on PHBV microsphere scaffold to regulate Hep3B cells activity and functionality: A model of liver tissue engineering system
Xin, H.Z. Seng, K.G. Wang, C.-H. Tong, Y.W.
Seng, K.G.
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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.
Keywords
Liver tissue engineering, Microsphere, PHBV, Protein, Surface conjugation
Source Title
Journal of Biomedical Materials Research - Part A
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Date
2007-11-01
DOI
10.1002/jbm.a.31257
Type
Article