通过多重流式细胞术分析龈沟液中炎症细胞因子的缓冲溶液优化
Optimization of buffer solutions to analyze inflammatory cytokines in gingival crevicular fluid by multiplex flow cytometry.
作者信息
Ríos-Lugo María-Judith, Martin Conchita, Alarcón José-Antonio, Esquifino Ana, Barbieri Germán, Solano Patricia, Sanz Mariano
机构信息
Rosalía de Castro 1, 1 izquierda, A Coruña. Spain,
出版信息
Med Oral Patol Oral Cir Bucal. 2015 Jan 1;20(1):e13-6. doi: 10.4317/medoral.19852.
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.
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.
摘要
目的
本研究旨在测试两种缓冲溶液，以便通过流式细胞术对龈沟液（GCF）中的炎性细胞因子（IL-1β、IL-6、TNF-α、骨保护素（OPG）、骨桥蛋白（OPN）和骨钙素（OC））进行可靠且可重复的分析。
材料与方法
用牙周试纸条收集健康志愿者的龈沟液样本，并在含有和不含蛋白酶抑制剂（PI）的磷酸盐缓冲盐水（PBS）或Tris-HCl缓冲液中进行稀释。通过流式细胞术（Luminex Xmap 200）进行细胞因子免疫测定并生成标准曲线。
结果
使用磷酸盐缓冲盐水（PBS）生成的标准曲线显示，在检测细胞因子IL-1ß、IL-6和TNF-α水平时，其与使用Tris-HCl时相比拟合度最佳（p<0.05）。
结论
添加PI的PBS缓冲液可对健康志愿者龈沟液样本中的炎性生物标志物进行可靠测量。
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