title: A dual-functional fibrous scaffold enhances P450 activity of cultured primary rat hepatocytes.
abstract: We have designed a novel dual-functional electrospun fibrous scaffold comprising two fiber mesh layers that were modified differently to induce two separate biological responses from hepatocytes. The first fiber layer was galactosylated on the surface to mediate hepatocyte attachment, while the second layer was loaded with 3-methylcholanthrene (3-Mc) to enhance cytochrome P450 activity of hepatocytes. Primary rat hepatocytes cultured on the galactosylated fibrous scaffolds loaded with different concentrations of 3-Mc were compared for their cell attachment efficiency, albumin secretion activity and cytochrome P450-dependent 7-ethoxycoumarin O-deethylase activity. This hybrid fibrous scaffold mediated hepatocyte attachment with slightly lower efficiency (76+/-2.3%) than a single-layer galactosylated fibrous scaffold (84+/-3.5%). More importantly, the cytochrome P450 activity of the hepatocytes cultured on the hybrid scaffold correlated well with the 3-Mc loading level. The results also showed that transfer of 3-Mc to hepatocytes through direct cell-fiber contact was the dominant transport route, with the induced cytochrome P450 activity being 1.9- to 4.8-fold higher than that of transfer of 3-Mc to hepatocytes via dissolution from fibers to medium. This study demonstrates the feasibility of creating multi-functional fibrous scaffolds that serve both as an adhesive substrate and as a delivery vehicle for bioactive molecules.
authors.affiliation: Division of Bioengineering and NUS Nanoscience and Nanotechnology Initiative, National University of Singapore, Singapore, Singapore.
authors.forename: Kian-Ngiap
authors.initials: KN
authors.lastname: Chua
authors.forename: Yen-Ni
authors.initials: YN
authors.lastname: Tang
authors.forename: Chai-Hoon
authors.initials: CH
authors.lastname: Quek
authors.forename: Seeram
authors.initials: S
authors.lastname: Ramakrishna
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.forename: Hai-Quan
authors.initials: HQ
authors.lastname: Mao
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005690:Galactose / Q000008:administration & dosage* / Q000737:chemistry; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008422:Materials Testing; D008748:Methylcholanthrene / Q000008:administration & dosage* / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D001672:Biocompatible Materials; D008748:Methylcholanthrene; D003577:Cytochrome P-450 Enzyme System; D005690:Galactose