title: Cellular Responses Modulated by FGF-2 Adsorbed on Albumin/Heparin Layer-by-Layer Assemblies.
abstract: In a typical cell culture system, growth factors immobilized on the cell culture surfaces can serve as a reservoir of bio-signaling molecules, without the need to supplement them additionally into the culture medium. In this paper, we report on the fabrication of albumin/heparin (Alb/Hep) assemblies for controlled binding of basic fibroblast growth factor (FGF-2). The surfaces were constructed by layer-by-layer adsorption of polyelectrolytes albumin and heparin and were subsequently stabilized by covalent crosslinking with glutaraldehyde. An analysis of the surface morphology by atomic force microscopy showed that two Alb/Hep bilayers are required to cover the surface of substrate. The formation of the Alb/Hep assemblies was monitored by the surface plasmon resonance (SPR), the infrared multiinternal reflection spectroscopy (FTIR MIRS) and UV/VIS spectroscopy. The adsorption of FGF-2 on the cross-linked Alb/Hep was followed by SPR. The results revealed that FGF-2 binds to the Alb/Hep assembly in a dose and time-dependent manner up to the surface concentration of 120 ng/cm(2). The bioactivity of the adsorbed FGF-2 was assessed in experiments in vitro, using calf pulmonary arterial endothelial cells (CPAE). CPAE cells could attach and proliferate on Alb/Hep surfaces. The adsorbed FGF-2 was bioactive and stimulated both the proliferation and the differentiation of CPAE cells. The improvement was more pronounced at a lower FGF-2 surface concentration (30 ng/cm(2)) than on surfaces with a higher concentration of FGF-2 (120 ng/cm(2)).
authors.affiliation: Department of Biomaterials and Bioanalogous Systems, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic.
authors.forename: Marta
authors.initials: M
authors.lastname: Kumorek
authors.affiliation: Department of Biomaterials and Bioanalogous Systems, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic.
authors.forename: Dana
authors.initials: D
authors.lastname: Kubies
authors.affiliation: Department of Biomaterials and Tissue Engineering, Institute of Physiology, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.
authors.forename: Elena
authors.initials: E
authors.lastname: Filová
authors.affiliation: Department of Biomaterials and Bioanalogous Systems, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic.
authors.forename: Milan
authors.initials: M
authors.lastname: Houska
authors.affiliation: Department of Biomaterials and Bioanalogous Systems, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic.
authors.forename: Naresh
authors.initials: N
authors.lastname: Kasoju
authors.affiliation: Department of Biomaterials and Bioanalogous Systems, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic.
authors.forename: Eliška
authors.initials: E
authors.lastname: Mázl Chánová
authors.affiliation: Department of Biomaterials and Tissue Engineering, Institute of Physiology, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic; Department of Medical Biophysics and Informatics, Third Faculty of Medicine, Charles University in Prague, Prague, Czech Republic; Department of Biomedical Technology, Faculty of Biomedical Engineering, Czech Technical University in Prague, Kladno, Czech Republic.
authors.forename: Roman
authors.initials: R
authors.lastname: Matějka
authors.affiliation: Department of Biomaterials and Tissue Engineering, Institute of Physiology, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.
authors.forename: Markéta
authors.initials: M
authors.lastname: Krýslová
authors.affiliation: Department of Biomaterials and Tissue Engineering, Institute of Physiology, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Bačáková
authors.affiliation: Department of Biomaterials and Bioanalogous Systems, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic.
authors.forename: František
authors.initials: F
authors.lastname: Rypáček
mesh_terms: D000418:Albumins / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D018625:Microscopy, Atomic Force; D011485:Protein Binding
chemical_list: D000418:Albumins; D003470:Culture Media; D016222:Fibroblast Growth Factor 2; D006493:Heparin