title: 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.
authors.affiliation: Department of Chemical and Biomolecular Engineering, National University of Singapore, 21 Lower Kent Ridge Road, Singapore 119077.
authors.forename: Xin Hao
authors.initials: XH
authors.lastname: Zhu
authors.forename: Seng Keat
authors.initials: SK
authors.lastname: Gan
authors.forename: Chi-Hwa
authors.initials: CH
authors.lastname: Wang
authors.forename: Yen Wah
authors.initials: YW
authors.lastname: Tong
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003432:Cross-Linking Reagents / Q000737:chemistry; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D006801:Humans; D008099:Liver / Q000166:cytology / Q000502:physiology; D008863:Microspheres; D008954:Models, Biological; D011091:Polyesters / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D013499:Surface Properties; D023822:Tissue Engineering; D000094122:Polyhydroxybutyrates
chemical_list: D003432:Cross-Linking Reagents; D005022:Ethyldimethylaminopropyl Carbodiimide; D016326:Extracellular Matrix Proteins; D011091:Polyesters; D013388:Succinimides; C052620:poly(3-hydroxybutyrate)-co-(3-hydroxyvalerate); D000094122:Polyhydroxybutyrates; C001426:N-hydroxysuccinimide