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Cell-protein-material Interactions on Bioceramics and Model Surfaces
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Surface Biotechnology.
2004 (English)Doctoral thesis, comprehensive summary (Other academic)Alternative title
Interaktioner mellan celler, proteiner och keramiska material (Swedish)
Abstract [en]

The objective of this thesis was to investigate and characterize the interaction between blood proteins and different surfaces with emphasis on protein adsorption to bioceramics and model surfaces. Special effort was made to monitor the spontaneous and selective adsorption of proteins from human plasma and to examine the orientation, conformation and functional behavior of single proteins after adsorption.

Five different ceramic biomaterials: alumina (Al2O3), zirconia (ZrO2), hydroxyapatite (Ca10(PO4)6(OH)2) and two glass-ceramics, AP40 (SiO2-CaO-Na2O-P2O5-MgO-K2O-CaF2) and RKKP (AP40 with Ta2O3-La2O3), were exposed to human plasma and their protein binding capacities and affinities for specific proteins were studied by chromatography, protein assays, two-dimensional gel electrophoresis and Western blotting. The studies showed that all materials adsorbed approximately the same high amount of plasma proteins and that they therefore should be fully covered by proteins in an in vivo setting. The adsorbed proteins were different for most materials which could explain their previously observed different levels of tissue integration in vivo.

Four of the proteins that behaved differently, ceruloplasmin, prothrombin, α2-HS-glycoprotein and α1-antichymotrypsin, were selected for characterization with atomic force microscopy and ellipsometry. The studies, which were performed on ultraflat silicon wafers (silica), showed that the proteins oriented themselves with their long axis parallel to the surface or as in case of ceruloplasmin with one of its larger sides towards the surface. All of them had globular shapes but other conformational details were not resolved. Furthermore, prothrombin (none of the others) formed multilayers at high proteins concentrations.

The functional behaviour of the adsorbed proteins, referring to their cell binding and cell spreading capacity on silica and a positive cell adhesion reference surface (Thermanox®), was affected by the underlying substrate. Ceruloplasmin, α2-HS-glycoprotein and α1-antichymotrypsin stimulated cell attachment to silica, but suppressed attachment to Thermanox®. Prothrombin stimulated cell attachment to both surfaces. The attachment was in most cases mediated both by cell membrane-receptors (integrins) and by non-specific interactions between the cell and the material.

This thesis showed that the compositional mixture, orientation, conformation and functional behavior of the adsorbed proteins are determined by the properties of the underlying surface and if these parameters are controlled very different cellular responses can be induced.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis , 2004. , p. 72
Series
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 1046
Keyword [en]
Chemistry, protein adsorption, blood proteins, biomaterials, ceramics, glass-ceramics, two-dimensional gel electrophoresis, protein orientation, silicon wafers, atomic force microscopy, bone cell adhesion, divalent cation dependency
Keyword [sv]
Kemi
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-4688ISBN: 91-554-6107-7 (print)OAI: oai:DiVA.org:uu-4688DiVA, id: diva2:165502
Public defence
2004-12-16, B42, BMC, Husargatan 3, Uppsala, 13:15
Opponent
Supervisors
Available from: 2004-11-24 Created: 2004-11-24Bibliographically approved
List of papers
1. Plasma protein adsorption pattern on characterized ceramic biomaterials
Open this publication in new window or tab >>Plasma protein adsorption pattern on characterized ceramic biomaterials
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2002 In: Biomaterials, ISSN 0142-9612, Vol. 23, p. 1237-1247Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-92427 (URN)
Available from: 2004-11-24 Created: 2004-11-24Bibliographically approved
2. Protein adsorption onto two bioactive glass-ceramics
Open this publication in new window or tab >>Protein adsorption onto two bioactive glass-ceramics
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2003 In: Biomaterials, ISSN 0142-9612, Vol. 24, p. 147-155Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-92428 (URN)
Available from: 2004-11-24 Created: 2004-11-24Bibliographically approved
3. Bone cell (MG63) adhesion on silicon wafers pre-coated with ceruloplasmin and prothrombin
Open this publication in new window or tab >>Bone cell (MG63) adhesion on silicon wafers pre-coated with ceruloplasmin and prothrombin
Article in journal (Refereed) Submitted
Identifiers
urn:nbn:se:uu:diva-92429 (URN)
Available from: 2004-11-24 Created: 2004-11-24Bibliographically approved
4. Characterization of adsorbed α2-HS-glycoprotein and α1-antichymotrypsin and their influence on bone cell (MG63) adhesion
Open this publication in new window or tab >>Characterization of adsorbed α2-HS-glycoprotein and α1-antichymotrypsin and their influence on bone cell (MG63) adhesion
Article in journal (Refereed) Submitted
Identifiers
urn:nbn:se:uu:diva-92430 (URN)
Available from: 2004-11-24 Created: 2004-11-24Bibliographically approved

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