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与聚合物材料接触凝固血液中的血清诱导的粒细胞呼吸爆发反应的不同动力学。

Different kinetics of the respiratory burst response in granulocytes, induced by serum from blood coagulated in contact with polymer materials.

作者信息

Nygren H, Braide M, Karlsson C

机构信息

Applied Cell Biology, Department of Anatomy and Cell Biology, University of Göteborg, Sweden.

出版信息

Biomaterials. 2000 Jan;21(2):173-82. doi: 10.1016/s0142-9612(99)00146-5.

Abstract

Tubes of different polymer materials were filled with blood collected by venous puncture. The blood was allowed to clot for 10 min, and the serum was collected. Complement activation was demonstrated through assessment of the C3-level by radial immunodiffusion. Phospholipid fingerprints were made after lipid extraction of serum and separation by thin-layer chromatography. The granulocyte fraction of venous blood was separated on a Percoll gradient and the cells were either loaded with a calcium probe, or incubated with luminol. These cells were used as a biological test for inflammatory mediators. Serum from blood coagulated in contact with different materials was added to the test cells. The intracellular calcium level was recorded by Calcium Green-1 fluorescence and the respiratory burst of the test cells was recorded by luminol-amplified chemiluminescence. Serum from blood coagulated in contact with glass tubes, methylised glass tubes and teflon (PTFE) tubes induced a transient increase of the cellular calcium level, indicating a G protein-coupled activation of the test cells. Serum from blood coagulated in contact with glass tubes, methylised glass tubes, and PTFE tubes primed the test cells for a subsequent f-MLP response. Serum from blood coagulated in contact with polyurethane and polypropylene induced a direct biphasic respiratory burst response in the test cells and serum from blood coagulated in contact with methylised glass induced a direct monophasic respiratory burst response in the test cells. Complement activation was demonstrated after blood contact with hydrophobic glass and PTFE. Different fingerprints of phospholipid content were found in sera after blood contact with different materials. The data show that different inflammatory mediators are released during blood coagulation in contact with different materials. The method may be valuable as a screening test for blood compatibility of materials.

摘要

将不同聚合物材料的试管装满通过静脉穿刺采集的血液。让血液凝固10分钟,然后收集血清。通过放射免疫扩散法评估C3水平来证明补体激活。血清进行脂质提取并通过薄层色谱分离后制作磷脂指纹图谱。在Percoll梯度上分离静脉血的粒细胞部分,这些细胞要么加载钙探针,要么与鲁米诺一起孵育。这些细胞用作炎症介质的生物学检测。将与不同材料接触凝固的血液的血清添加到测试细胞中。通过钙绿-1荧光记录细胞内钙水平,通过鲁米诺放大的化学发光记录测试细胞的呼吸爆发。与玻璃管、甲基化玻璃管和聚四氟乙烯(PTFE)管接触凝固的血液的血清会引起细胞钙水平的短暂升高,表明测试细胞发生了G蛋白偶联激活。与玻璃管、甲基化玻璃管和PTFE管接触凝固的血液的血清使测试细胞对随后的f-MLP反应产生预激发。与聚氨酯和聚丙烯接触凝固的血液的血清在测试细胞中诱导直接的双相呼吸爆发反应,与甲基化玻璃接触凝固的血液的血清在测试细胞中诱导直接的单相呼吸爆发反应。血液与疏水玻璃和PTFE接触后证明有补体激活。血液与不同材料接触后,血清中发现了不同的磷脂含量指纹图谱。数据表明,在血液与不同材料接触凝固过程中会释放不同的炎症介质。该方法作为材料血液相容性的筛选测试可能具有价值。

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