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Initial in vitro interaction of osteoblasts with nano-porous alumina
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Surface Biotechnology.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Surface Biotechnology, Centre for Surface Biotechnology.
2003 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 24, no 18, 3039-3046 p.Article in journal (Refereed) Published
Abstract [en]

In the present study we have used a characterised primary human cell culture model to investigate cellular interactions with nano-porous alumina. This material, prepared by anodisation, is being developed as a coating on titanium alloy implants. The structure of the alumina, as determined by X-ray diffraction and transmission electron microscopy, was amorphous. When studying cell/material interactions we used both biochemical and morphological parameters. Cell viability, proliferation and phenotype were assessed by measurement of redox reactions in the cells, cellular DNA, tritiated thymidine ([H-3]-TdR) incorporation and alkaline phosphatase (ALP) production. Results showed a normal osteoblastic growth pattern with increasing cell numbers during the first 2 weeks. A peak in cell proliferation was seen on day 3, after which cell growth decreased, followed by an increase in ALP production, thus indicating that the osteoblastic phenotype was retained on the alumina. Cell adhesion was observed, the osteoblast-like cells having a flattened morphology with filipodia attached to the pores of the material. SDS-PAGE and western blot measurements showed that the nano-porous alumina was able to adsorb fibronectin. Trace amounts of aluminium ions were measured in the surrounding medium, but no adverse effect on cell activity was observed.

Place, publisher, year, edition, pages
2003. Vol. 24, no 18, 3039-3046 p.
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:uu:diva-91966DOI: 10.1016/S0142-9612(03)00146-7ISI: 000183097900008OAI: oai:DiVA.org:uu-91966DiVA: diva2:164886
Available from: 2004-09-01 Created: 2004-09-01 Last updated: 2014-01-28Bibliographically approved
In thesis
1. Nano-porous Alumina, a Potential Bone Implant Coating
Open this publication in new window or tab >>Nano-porous Alumina, a Potential Bone Implant Coating
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis describes a method of growing a highly adherent nano-porous alumina coating on titanium implant materials, a design which might be useful in hard tissue replacement. Alumina layers were formed by anodisation of aluminium, which had been deposited on titanium and titanium alloys by electron beam evaporation. Mechanical testing showed the coatings’ shear and tensile strength to be ~20MPa and ~10MPa respectively.

Human osteoblasts were cultured on purchased membranes, produced in the same way with similar characteristics as the coating mentioned above. Cell viability, proliferation and phenotype were assessed by measuring redox reactions, DNA, tritiated thymidine incorporation and alkaline phosphatase production. Results showed normal osteoblastic growth patterns with increasing cell numbers the first two weeks after which cell growth decreased and alkaline phosphatase production increased, indicating that osteoblastic phenotype was retained on the alumina. Flattened cell morphology with filipodia attached to the pores of the material was seen.

Implants frequently trigger inflammatory responses due to accumulation and activation of cells such as polymorphonuclear granulocytes (PMN), also called neutrophils. Activation and morphology of human PMN in response to nano-porous alumina with two pore sizes (20 and 200 nm) was investigated by luminol-amplified chemiluminescence, granule enzyme deposition measurement, optical and scanning electron microscopy. Activation was observed on both membrane types, however less pronounced on the 200 nm alumina. For both membranes a decrease in activation was seen after coating with fibrinogen, collagen I and serum (more pronounced for the two latter). On fibrinogen-coated alumina many flattened cells were observed, indicating frustrated phagocytosis. Finally when culturing osteoblasts on non-coated and collagen-coated membranes (after exposure to PMN) many more cells had established on the protein-coated surface after 24 h.

The overall results indicate that it might be possible to produce a novel bone implant coating by anodisation of aluminium deposited on titanium and that this material will support osteoblast adhesion and proliferation. Furthermore neutrophil activation can be suppressed when coating the alumina with collagen I, which is beneficial considering the fact that this protein also is essential for bone formation.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2004. 75 p.
Series
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 997
Keyword
Chemistry, Nano-porous aluminium oxide, Biomaterial, Hard tissue implant, Surface topography, Osteoblast, Biocompatibility, Neutrophil, Inflammation, Kemi
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-4452 (URN)91-554-6007-0 (ISBN)
Public defence
2004-09-29, B-42, BMC, Husargatan 3, Uppsala, 09:15
Opponent
Supervisors
Available from: 2004-09-01 Created: 2004-09-01Bibliographically approved

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