Optimization of buffer solutions to analyze infammatory cytokines in gingival crevicular fuid by multiplex fow cytometry
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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 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. © Medicina Oral S. L.
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Crevicular fuid; Curve ftting; Cytokines; Fow cytometer; Immunoassay buffer biological marker; cytokine; interleukin 1beta; interleukin 6; osteopontin; osteoprotegerin; phosphate buffered saline; proteinase inhibitor; trometamol; tumor necrosis factor alpha; buffer; cytokine; Article; controlled study; curve fitting; dilution curve; flow cytometry; gingivitis; human; immunoassay; normal human; protein binding; reaction optimization; room temperature; sensitivity analysis; test strip; chemistry; comparative study; flow cytometry; gingivitis; inflammation; Buffers; Cytokines; Flow Cytometry; Gingival Crevicular Fluid; Humans; Inflammation
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