Titanium particles activate toll-like receptor 4 independently of lipid rafts in RAW264.7 murine macrophages.
Titanium particles activate toll-like receptor 4 independently of lipid rafts in RAW264.7 murine macrophages.
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DOI：
10.1002/jor.21199
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发表时间：
2011-02
影响因子：
2.8
通讯作者：
Greenfield, Edward M.
中科院分区：
文献类型：
--
作者：
Islam, Andrew S.;Beidelschies, Michelle A.;Huml, Anne;Greenfield, Edward M.
粘附性PAMP（病原体相关分子模式）通过Toll样受体2（TLR 2）和TLR 4发挥作用，以增加细胞培养和植入物松动动物模型中骨科磨损颗粒的生物活性。这项研究测试了这是否依赖于TLR与脂筏的结合，如对可溶性TLR配体的反应所报道的那样。为此，通过暴露于具有粘附的PAMP、可溶性脂多糖（LPS）、可溶性脂磷壁酸（LTA）或热灭活细菌的钛颗粒来活化RAW264.7鼠巨噬细胞，所述热灭活细菌已经被广泛洗涤以去除可溶性PAMP。通过两种独立的方法分离脂筏，并通过蛋白质印迹法分析TLR 4和TLR 2的位置。当用可溶性LPS和LTA激活巨噬细胞时，同源TLR与脂筏相关，但在用粘附PAMP的钛颗粒或热灭活细菌刺激后则不相关。脂筏破坏剂甲基-β-环糊精剂量依赖性地抑制LPS引起的TNFα释放，但对粘附PAMP的钛颗粒引起的TNFα释放无影响。因此，我们得出结论，钛颗粒与粘附PAMPs和热灭活细菌激活TLR 2和TLR 4在巨噬细胞中，而不诱导TLR与脂筏。这些结果对骨科植入物松动的机制以及其他炎症情况下TLR激活的机制具有重要意义。
Adherent PAMPs (pathogen-associated molecular patterns) act through Toll-like receptor2 (TLR2) and TLR4 to increase the biological activity of orthopaedic wear particles in cell culture and animal models of implant loosening. This study tested whether this is dependent on TLR association with lipid rafts as reported for the response to soluble TLR ligands. For this purpose, RAW264.7 murine macrophages were activated by exposure to titanium particles with adherent PAMPs, soluble lipopolysaccharide (LPS), soluble lipotecichoic acid (LTA), or heat-killed bacteria that had been extensively washed to remove soluble PAMPs. Lipid rafts were isolated by two independent methods and the location of TLR4 and TLR2 was analyzed by western blotting. The cognate TLRs associated with lipid rafts when the macrophages were activated with soluble LPS and LTA but not after stimulation with either titanium particles with adherent PAMPs or heat-killed bacteria. The lipid raft disruptor, methyl-β-cyclodextrin, dose-dependently inhibited TNFα release in response to LPS but had no affect on TNFα release in response to titanium particles with adherent PAMPs. We conclude, therefore, that titanium particles with adherent PAMPs and heat-killed bacteria activate TLR2 and TLR4 in macrophages without inducing either TLR to associate with lipid rafts. These results have important implications for the mechanisms of orthopaedic implant loosening as well the mechanisms for TLR activation in other inflammatory situations.
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影响因子：
2.8
作者：
Tatro, Joscelyn M.;Taki, Naoya;Greenfield, Edward M.
通讯作者：
Greenfield, Edward M.
影响因子：
4.8
作者：
Triantafilou, M;Manukyan, M;Triantafilou, K
通讯作者：
Triantafilou, K
影响因子：
2.8
作者：
Taki, N;Tatro, JM;Greenfield, EM
通讯作者：
Greenfield, EM
DOI：
10.1080/00016470310018135
发表时间：
2003-12-01
期刊：
ACTA ORTHOPAEDICA SCANDINAVICA
影响因子：
--
作者：
Engesæter, LB;Lie, SA;Havelin, LI
通讯作者：
Havelin, LI
影响因子：
4.9
作者：
Takagi, Michiaki;Tamaki, Yasunobu;Salo, Jari
通讯作者：
Salo, Jari