title: Optimization of buffer solutions to analyze inflammatory cytokines in gingival crevicular fluid by multiplex flow cytometry.
abstract: 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.
  
  
  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.
authors.affiliation: Rosalía de Castro 1, 1 izquierda, A Coruña. Spain, germanbarbieri@gmail.com.
authors.forename: María-Judith
authors.initials: MJ
authors.lastname: Ríos-Lugo
authors.forename: Conchita
authors.initials: C
authors.lastname: Martin
authors.forename: José-Antonio
authors.initials: JA
authors.lastname: Alarcón
authors.forename: Ana
authors.initials: A
authors.lastname: Esquifino
authors.forename: Germán
authors.initials: G
authors.lastname: Barbieri
authors.forename: Patricia
authors.initials: P
authors.lastname: Solano
authors.forename: Mariano
authors.initials: M
authors.lastname: Sanz
mesh_terms: D002021:Buffers; D016207:Cytokines / Q000032:analysis*; D005434:Flow Cytometry; D005883:Gingival Crevicular Fluid / Q000737:chemistry*; D006801:Humans; D007249:Inflammation
chemical_list: D002021:Buffers; D016207:Cytokines