...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >Gene monitoring of surface-activated monocytes in circulating whole blood using duplex RT-PCR
【24h】

Gene monitoring of surface-activated monocytes in circulating whole blood using duplex RT-PCR

机译：使用双重RT-PCR监测循环全血中表面活化单核细胞的基因

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Previous studies have shown that blood-contacting materials activate blood monocytes. The aim of this study is to evaluate a highly sensitive technique for detecting changes at the mRNA level in circulating monocytes and to find suitable "gene markers" for assessing the hemocompatibility of biomaterials. Human blood was recir-culated in a modified Chandler Loop formed of test tubes. Five groups of biomaterials were compared. Monocytes were separated by using magnetic beads, and gene expression analyses were performed using RT-PCR. We investigated the mRNA expression of stress proteins (hsp70) and inflammatory markers (IL-1#alpha#, IL-1#beta#, IL-6, TNF-#alpha#). mRNAs for cytokines were highly upregulated in LPS-stimulated monocytes. Hsp70 transcripts were upregulated after heat shock but were not influenced by treatment with LPS. The gene expression of the cytokines was affected by various biomaterials. The intensity of the inflammatory response increased in the following order: heparin coatings (PVC) < uncoated PVC < silicone, polypeptide-coated PVC. No change was seen in the expression of the hsp70 genes. An inhibitory effect of systemic heparin on surface-activated monocytes was observed for the mRNA expression of the cytokines. The recirculation of human blood in an in vitro model in combination with the immunomagnetic separation of monocytes and the Duplex RT-PCR method is a powerful tool for getting reliable results. Our data demonstrate that hsp70 genes cannot be recommended for short-term, biomaterial-induced mRNA expression studies. Conversely, mRNAs for IL-1#alpha#, IL-1#beta#, IL-6, and TNF-#alpha# were expressed in a material-dependent manner. Thus our model provides an effective tool for assessing the hemocompatibility of biomaterials before their clinical application and it also can serve as a safety control during the industrial manufacturing process. This method can be applied to various blood cell populations and numerous gene expression studies and may enable a more fundamental understanding of the biologic processes involved in blood-material interactions.
机译：先前的研究表明，血液接触材料会激活血液单核细胞。这项研究的目的是评估一种高度灵敏的技术，以检测循环单核细胞中mRNA水平的变化，并找到合适的“基因标记”来评估生物材料的血液相容性。在由试管形成的改进的钱德勒环中循环人类血液。比较了五组生物材料。通过使用磁珠分离单核细胞，并使用RT-PCR进行基因表达分析。我们调查了应激蛋白（hsp70）和炎症标志物（IL-1＃alpha＃，IL-1＃beta＃，IL-6，TNF-＃alpha＃）的mRNA表达。 LPS刺激的单核细胞中细胞因子的mRNA高度上调。热休克后Hsp70转录本上调，但不受LPS处理的影响。细胞因子的基因表达受到多种生物材料的影响。炎症反应的强度按以下顺序增加：肝素涂层（PVC）<未涂层的PVC <硅酮，多肽涂层的PVC。 hsp70基因的表达未见变化。对于细胞因子的mRNA表达，观察到全身性肝素对表面活化的单核细胞的抑制作用。体外模型中人血的再循环结合单核细胞的免疫磁分离和Duplex RT-PCR方法是获得可靠结果的有力工具。我们的数据表明，hsp70基因不能推荐用于短期，生物材料诱导的mRNA表达研究。相反，IL-1＃alpha＃，IL-1＃beta＃，IL-6和TNF-＃alpha＃的mRNA以材料依赖的方式表达。因此，我们的模型为在生物材料临床应用之前评估其血液相容性提供了有效的工具，并且还可以用作工业生产过程中的安全控制。该方法可以应用于各种血细胞群体和大量的基因表达研究，并且可以使人们更深入地了解涉及血液-物质相互作用的生物学过程。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号