Optimization of buffer solutions to analyze infammatory cytokines in gingival crevicular fuid by multiplex fow cytometry

Abstract

Objective: the aim of this study was to test two buffer solutions in order to attain a reliable and reproducible analysis of infammatory cytokines (IL-1β, IL-6, TNF-α, OPG, OPN and OC), in gingival crevicular fuid (GCF) by fow 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 fow cytometry (Luminex Xmap200) 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 infammatory bi-omarkers in GCF samples of healthy volunteers.