Optimization of buffer solutions to analyze inflammatory cytokines in gingival crevicular fluid by multiplex flow cytometry

  1. María Judith Ríos Lugo
  2. Conchita Martín
  3. Jose Antonio Alarcón
  4. Ana Isabel Esquifino Parras
  5. Germán Barbieri Petrelli
  6. Patricia Solano
  7. Mariano Sanz Alonso
Medicina oral, patología oral y cirugía bucal. Ed. inglesa

ISSN: 1698-6946

Year of publication: 2015

Volume: 20

Issue: 1

Pages: 5

Type: Article

DOI: 10.4317/MEDORAL.19852 DIALNET GOOGLE SCHOLAR lock_openOpen access editor

More publications in: Medicina oral, patología oral y cirugía bucal. Ed. inglesa


Objective: the aim of this study was to test two buffer solutions in order to attain a reliable and reproducible analysis of inflammatory cytokines (IL-1β, IL-6, TNF-α, OPG, OPN and OC), in gingival crevicular fluid (GCF) by flow cytometry. Material and Methods: GCF samples from healthy volunteers were collected with perio-paper strips and diluted either in phosphate buffered saline (PBS) or Tris-HCl buffer, with and without protease inhibitors (PI). Cytokine immunoassays were carried out by flow cytometry (Luminex Xmap 200 ) generating standard curves. Results: standards curves generated with the use of phosphate-buffered saline (PBS) demonstrated best adjustment for cytokines IL-1ß, IL-6 and TNF- α levels, when using Tris-HCl ( p <0.05). Conclusions: The use of PBS buffer with the addition of PI provided reliable measurements of inflammatory biomarkers in GCF samples of healthy volunteers.