title: Proteomic analysis of human osteoprogenitor response to disordered nanotopography.
abstract: Previous studies have shown that microgroove-initiated contact guidance can induce bone formation in osteoprogenitor cells (OPGs) and produce changes in the cell proteome. For proteomic analysis, differential in-gel electrophoresis (DIGE) can be used as a powerful diagnostic method to provide comparable data between the proteomic profiles of cells cultured in different conditions. This study focuses on the response of OPGs to a novel nanoscale pit topography with osteoinductive properties compared with planar controls. Disordered near-square nanopits with 120 nm diameter and 100 nm depth with an average 300 nm centre-to-centre spacing (300 nm spaced pits in square pattern, but with +/-50 nm disorder) were fabricated on 1x1 cm2 polycaprolactone sheets. Human OPGs were seeded onto the test materials. DIGE analysis revealed changes in the expression of a number of distinct proteins, including upregulation of actin isoforms, beta-galectin1, vimentin and procollagen-proline, 2-oxoglutarate 4-dioxygenase and prolyl 4-hydroxylase. Downregulation of enolase, caldesmon, zyxin, GRASP55, Hsp70 (BiP/GRP78), RNH1, cathepsin D and Hsp27 was also observed. The differences in cell morphology and mineralization are also reported using histochemical techniques.
authors.affiliation: Division of Infection and Immunity, Centre for Cell Engineering, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow, Glasgow G12 8QQ, UK. 0506972k@student.gla.ac.uk
authors.forename: Fahsai
authors.initials: F
authors.lastname: Kantawong
authors.forename: Richard
authors.initials: R
authors.lastname: Burchmore
authors.forename: Nikolaj
authors.initials: N
authors.lastname: Gadegaard
authors.forename: Richard O C
authors.initials: RO
authors.lastname: Oreffo
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Dalby
mesh_terms: D019295:Computational Biology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D000091342:Endoplasmic Reticulum Chaperone BiP; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006651:Histocytochemistry; D006801:Humans; D008855:Microscopy, Electron, Scanning; D049329:Nanostructures; D010012:Osteogenesis / Q000502:physiology*; D011091:Polyesters; D040901:Proteomics / Q000379:methods; D013234:Stem Cells / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D011091:Polyesters; C016240:polycaprolactone