Background: Gingival crevicular fluid (GCF) may be a source of new biomarkers of periodontitis/gingivitis.However, the minute volumes of GCF harvested in healthy sites are a serious drawback. We evaluated howpre-analytical and analytical variables concerning GCF collection and processing, could significantly influencequality and reproducibility of MALDI-TOF profiles.Methods: GCF was collected from healthy sites. The use of paper strips vs paper points was compared. Peptidesand proteins were extracted by centrifugal elution in different solutions, at different accelerations, with andwithout protease inhibitor cocktail (PIC). Finally, we evaluated how matrix composition and matrix/samplevolume ratio affect the reproducibility of MALDI-TOF profiles.Results: Trifluoroacetic acid elution generated richer gingival fingerprints compared to acetic acid, independentlyof the collection device. Centrifugation speed and PIC supplementation did not change GCF profiles. A finemodulation of matrix composition and matrix/sample volume ratio resulted in a satisfactory reproducibility(CV less than 10% for peak area and signal-to-noise ratio).Conclusion: An optimized procedure, enabling generation of reproducible MALDI-TOF profiles from limitedvolume of GCF, is proposed. These fingerprints may serve as reference for future studies oriented to the maintenanceand preservation of good gingival status and to discovery biomarkers of periodontitis/gingivitis.

Assessment of pre-analytical and analytical variables affecting peptidome profiling of gingival crevicular fluid by MALDI-TOF mass spectrometry

Preianò M;Falcone D;Maggisano G;Montalcini T;Paduano S;Savino R;TERRACCIANO R
2014-01-01

Abstract

Background: Gingival crevicular fluid (GCF) may be a source of new biomarkers of periodontitis/gingivitis.However, the minute volumes of GCF harvested in healthy sites are a serious drawback. We evaluated howpre-analytical and analytical variables concerning GCF collection and processing, could significantly influencequality and reproducibility of MALDI-TOF profiles.Methods: GCF was collected from healthy sites. The use of paper strips vs paper points was compared. Peptidesand proteins were extracted by centrifugal elution in different solutions, at different accelerations, with andwithout protease inhibitor cocktail (PIC). Finally, we evaluated how matrix composition and matrix/samplevolume ratio affect the reproducibility of MALDI-TOF profiles.Results: Trifluoroacetic acid elution generated richer gingival fingerprints compared to acetic acid, independentlyof the collection device. Centrifugation speed and PIC supplementation did not change GCF profiles. A finemodulation of matrix composition and matrix/sample volume ratio resulted in a satisfactory reproducibility(CV less than 10% for peak area and signal-to-noise ratio).Conclusion: An optimized procedure, enabling generation of reproducible MALDI-TOF profiles from limitedvolume of GCF, is proposed. These fingerprints may serve as reference for future studies oriented to the maintenanceand preservation of good gingival status and to discovery biomarkers of periodontitis/gingivitis.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12317/10281
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