Optimization of buffer solutions to analyze inflammatory cytokines in gingival crevicular fluid by multiplex flow cytometry
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Ríos Lugo, María Judith; Martín, Conchita; Alarcón, Jose Antonio; Esquifino Parras, Ana Isabel; Barbieri, Germán; Solano, Patricia; Sanz Alonso, Mariano
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Aquest document és un/a article, creat/da en: 2015
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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.
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