Effect of Co2+ and Cr3+ as artificial joint wear product on
tumor necrosis factor secreted from human mononuclear cells*★

Abstract

AIM： To discuss the effect of artificial joint wear production meal ion cobalt (Co2+) and chromium (Cr3+) on aseptic loosening of artificial joint.

METHODS：The experiment was accomplished from June 2005 to January 2006 in the Burn Laboratory of the First Hospital Affiliated to Nanchang University.①CoCl2 powder and CrCl3 powder were dissolved in asepsis injection water for preparing the corresponding solutions, and confirmed with endotoxin before use.②Lymphofolt and Percoll gradient centrifugation were used to acquire mononuclear cell from peripheral blood, then the cells were cultured in the 24-hole plate, with 4×106 cell per hole.③CoCl2 solution and CrCl3 solution were added into monocyte liquid to adjust their final concentration to 1×10－5 and 5×10－4, while mononuclear cells cultured alone were served as negative.④After cells were incubated for 12, 24 and 48 hours, ELISA method was applied to detect tumor necrosis factor (TNF) α content in serum supernatant via the absorbance (A)

RESULT: ①When monocytes were exposed to Co2+ for 12, 24 and 48 hours, A value of serum supernatant was 0.636 2±0.299 3, 1.091 5±0.408 1, 0.715 0±0.295 1. TNF α content was obviously higher at hour 24 than at hours 12 and 48.②When monocytes were co-cultured with Cr3+, A value arrived the peak at hour 24 and declined at hour 48 (0.493 7±0.116 5, 0.872 7±0.280 1, 0.508 8±0.231 0 at hours 12, 24 and 48).

CONCLUSION:Co2+ and Cr3+ can stimulate human mononuclear cells release TNF α and induce osteolysis, resulting in aseptic loosening of artificial joint.

摘要
目的：观察人工关节磨损产物金属离子钴（Co2+）、铬(Cr3+)对人工关节无菌性松动的影响。
方法：试验于2005－06/2006－01在南昌大学第一附属医院烧伤实验室完成。①将CoCl2粉末与CrCl3粉末溶于无菌注射用水配制成CoCl2溶液和CrCl3溶液，使用前内毒素鳌试剂检测均证实无内毒素。②用lymphoflot和Percoll梯度离心法获得志愿者外周血中的单核细胞，将细胞在24孔培养皿中培养，4×106个/孔。③将CoCl2溶液和CrCl3溶液分别加入人单核细胞液中，使其终末溶度分别为1×10-5和5×10-4, 单核细胞单独培养作为对照组。④分别于12，24，48 h取细胞上清液，用ELISA法检测上清液中肿瘤坏死因子含量，以吸光度（A值）的高低来反映标本中肿瘤坏死因子α含量的多少。
结果： ①Co2+刺激单核细胞分泌肿瘤坏死因子结果：Co2+与人单核细胞共培养后，12，24，48 h检测上清液A值分别为0.636 2± 0.299 3，1.091 5±0.408 1，0.715 0±0.295 1，实验组24 h肿瘤坏死因子含量明显高于12 h和48 h肿瘤坏死因子含量。②Cr3+刺激单核细胞分泌肿瘤坏死因子结果：Cr3+与人单核细胞共培养后，12，24，48 h上清液A值分别为0.493 7±0.116 5，0.872 7±0.280 1，0.508 8±0.231 0，与Co2+类似,与Cr3+共培养后单核细胞上清液A值在24 h达到峰值，48 h又下降。
结论： 金属离子Co2+，Cr3+能通过刺激人单核细胞释放肿瘤坏死因子，诱发假体周围骨溶解，进而引起假体松动。
关键词：金属； 人工关节；肿瘤坏死因子类

郝亮，戴闽，帅浪，程涛，范红先，宗世璋.人工关节磨损产物金属离子钴、铬对人单核细胞分泌肿瘤坏死因子的影响[J].中国组织工程研究与临床康复，2007，11(1):67-69 [www.zglckf.com/zglckf/ejournal/upfiles/07-1/1k-67(ps).pdf]