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Optimization of buffer solutions to analyze inflammatory cytokines in gingival crevicular fluid by multiplex flow
María-Judith Ríos-Lugo1, Conchita Martin, José-Antonio Alarcón
1Rosalía de Castro 1, 1 izquierda, A Coruña. Spain, germanbarbieri@gmail.com.
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.
