| 1297 Crevicular fluid protein identification from orthodontic patients | ||
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V. WEISE1, M. BRICKLEY1, C. PENFOLD1, and J. SHEPHERD2, 1Orthodontic Biology Research Group, Yeovil, Somerset, United Kingdom, 2University of Wales College of Medicine, Cardiff, Wales, Uk Objectives: - To investigate the utility of vertical SDS-PAGE electrophoresis for the isolation and identification of proteins present in gingival crevicular fluid (GCF) during orthodontic treatment. - To identify key GCF proteins present during tooth movement which warrant investigation as potential biomarkers. Method: Multiple GCF samples (n=8) were collected from the upper arches of 30 juvenile (13 to 18 years old) patients (good oral hygiene, no periodontal inflammation) undergoing fixed orthodontic treatment using sterile paper points. Single patient samples were examined both individually (n=2) and pooled (n=6). Samples were eluted into sterile saline (30µl) prior to being subjected to SDS-PAGE electrophoresis. Silver staining and colloidal comassie blue staining were used to detect the resultant polypeptide bonds and were analysed using relative molecular weight determination software. Results: GCF proteins were successfully isolated using vertical SDS-PAGE electrophoresis. GCF samples from single sites resolved into a mean of 5 bands compared with pooled samples which provided significantly better resolution (mean 15 bands). Colloidal comassie blue staining provided comparable resolution to silver staining at low cost and with no use of toxic reagents. Prominent GCF polypeptides were seen at 79, 43 and 27.5kDa as well as at 60kDa representing alkaline phosphatase. Conclusion: The pooling of multiple GCF samples allowed a wide range GCF proteins to be isolated using vertical SDS-PAGE electrophoresis while single samples still resolved into several proteins present at higher concentrations. Refinement of the techniques used would potentially permit single-site sampling of key biomarkers allowing examination of up-regulation of these during orthodontic tooth movement. It is concluded that SDS-PAGE is a useful technique to apply to analysis of GCF biomarkers in orthodontic patients and further refinement of this may lead to the development of clinically relevant assays of GCF biomarkers in orthodontic patients. | ||
| Seq #143 - Factors That Respond to or Influence Orthodontic Tooth Movement 2:00 PM-4:00 PM, Thursday, 10 March 2005 Baltimore Convention Center Exhibit Hall E-F | ||
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