The role of adsorbed endotoxin in particle-induced stimulation of cytokine release
The role of adsorbed endotoxin in particle-induced stimulation of cytokine release
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DOI：
10.1016/s0736-0266(01)00179-6
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发表时间：
2002-07-01
影响因子：
2.8
通讯作者：
Goldring, SR
中科院分区：
文献类型：
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作者：
Cho, DR;Shanbhag, AS;Goldring, SR
许多体外模型已经证明磨损颗粒能够刺激可溶性促炎产物的释放，并能够诱导局部骨吸收。最近的观察表明，脂多糖（LPS）与颗粒磨损碎片的结合可以显着调节体外模型中的细胞响应模式。这些发现引起了人们对 LPS 在全关节置换术后无菌性松动发病机制中可能发挥的作用的担忧，并且还表明了在用于研究磨损碎片反应性质的体外模型中控制 LPS 污染可能产生的混杂影响的重要性。我们的研究旨在严格分析颗粒相关 LPS 对细胞反应的影响，并评估不同治疗方案对不同颗粒材料相关 LPS 的灭活效果。钴铬合金、钛-6-铝4-钒、氮化钛和二氧化硅颗粒用LPS进行预处理，并接受多种处理方案。当用乙醇洗涤制备的“原样”颗粒处理细胞时，检测到少量的 TNF-α、IL-1β 和 IL-1α。相比之下，所有用 LPS 预处理的颗粒种类即使在乙醇洗涤后，TNF-α、IL-1α 和 IL-1β 的释放也显着增加，并且相应的 mRNA 水平上调。在 1% 乙酸中煮沸 LPS 预处理的颗粒或高压灭菌并烘烤颗粒也会显着降低甚至在某些情况下消除 LPS 预处理的效果。这表明 LPS 与不同成分的颗粒表面结合，并且结合的 LPS 具有生物活性。灭活颗粒相关脂多糖的治疗方案显示出疗效的显着差异。当采用最严格的治疗时，基本上所有 LPS 活性都可以被消除。用这些方法处理的颗粒保留了一些刺激细胞因子释放的能力，但活性显着降低。这些结果提供了进一步的证据，表明LPS污染颗粒材料可以显着增强其生物活性。这种潜在的混杂效应需要在用于评估磨损颗粒的体外模型系统中仔细监测和控制。此外，体内骨植入物界面处颗粒相关内毒素的存在可以显着增强颗粒磨损碎片的不利生物活性。 (C) 2002 年由 Elsevier Science Ltd. 代表骨科研究学会出版。
Numerous in vitro models have demonstrated the capacity of wear particles to stimulate the release of soluble pro-inflammatory products with the ability to induce local bone resorption. Recent observations have demonstrated that binding of lipopolysaccharide (LPS) to particulate wear debris can significantly modulate the pattern of cell response in the in vitro models. These findings raise concerns over the possible role of LPS in the pathogenesis of aseptic loosening after total joint replacements, and also indicates the importance of controlling for possible confounding effects of LPS contamination in the in vitro models used to study the reactive nature of wear debris. Our studies were undertaken to rigorously analyze the effects of particle-associated LPS on cell responses and to assess the efficacy of different treatment protocols to inactivate LPS associated with different particulate materials. Particles of cobalt-chrome alloy, titanium-6-aluminum 4-vanadium, titanium nitride and silica were pretreated with LPS and exposed to multiple treatment protocols. When cells were treated with "as-received" particles prepared by washing in ethanol, small amounts of TNF-alpha, IL-1beta, and IL-1alpha were detected. In contrast, all particle species pretreated with LPS produced marked increases in TNF-alpha, IL-1alpha, and IL-1beta release, as well as upregulation of corresponding mRNA levels even after ethanol washing. Boiling the LPS-pretreated particles in 1% acetic acid or autoclaving and baking the particles also markedly reduced and in some instances abolished the effect of the LPS-pretreatment. This indicates that LPS binds to the surface of particles of diverse composition and that the bound LPS is biologically active. Treatment protocols to inactivate particle-associated LPS demonstrated significant differences in efficacy. When the most rigorous treatments were utilized, essentially all LPS activity could be eliminated. Particles treated with these methods retained some capacity to stimulate cytokine release, but activities were markedly reduced. These results provide further evidence indicating that LPS contamination of particulate materials can markedly enhance their biological activity. This potential confounding effect needs to be carefully monitored and controlled in the in vitro model systems used to evaluate wear particles. Furthermore, the presence of particle-associated endotoxin at the bone implant interface in vivo could markedly enhance the adverse biological activity of particulate wear debris. (C) 2002 Published by Elsevier Science Ltd. on behalf of Orthopaedic Research Society.