Xiong Jianwei, Dai Min, Hao Liang, Qi Qihua, Zhan Ping
Department of Orthopedics, First Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, PR China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2009 Mar;23(3):299-302.
To observe the human mononuclear cell releasing TNF-alpha and the activation of Caspase-3 during apoptosis after stimulated by Co2+ and Cr3+, to discuss the mechanism of artificial joint wear production metal ion on aseptic loosening.
CoCl2 powder and CrCl3 powder were dissolved by asepsis inject water, preparing solution for 10 mg/L and 500 mg/L, respectively. Mononuclear cells were acquired from peripheral blood, 4 x 10(6)/culture dish. According to the difference of culture solution, the cells were divided into 3 groups. Group A: mononuclear cell was culture with normal saline as control; group B: mononuclear cell was cultured with CoC2 solution; group C: mononuclear cell was cultured with CrCl3 solution. The production of TNF-alpha was assessed by ELISA, the activation of Caspase-3 was measured by colorimetric assay and the apoptotic cell was detected by TUNEL assays at 12, 24 and 48 hours after co-cultured respectively.
The concentration of TNF-alpha and the expression of Caspase-3 in groups B and C were higher than those in group A (P < 0.05), and reached the peak level at 24, 48 hours, respectively. The TUNEL positive cells were detected in the all groups, nucleus was pyknotic and darker-staining, cell body was crinkle and cell membrane was integrity. There were significant differences in the apoptosis rate between groups B, C and group A (P < 0.05). And the activation of Caspase-3 increased and had the positive correlation with the apoptosis rate (r = 0.765).
Co2+ and Cr3+ ions can stimulate human mononuclear cell to release TNF-alpha and induce human mononuclear cell apoptosis, which result in periprosthetic osteolysis and related to activation of Caspase-3.
观察Co2+和Cr3+刺激后人单核细胞释放肿瘤坏死因子-α（TNF-α）及凋亡过程中半胱天冬酶-3（Caspase-3）的激活情况，探讨人工关节磨损产生的金属离子导致无菌性松动的机制。
将CoCl2粉末和CrCl3粉末用无菌注射用水溶解，分别配制10mg/L和500mg/L的溶液。从外周血获取单核细胞，4×10(6)/培养皿。根据培养液不同将细胞分为3组。A组：单核细胞用生理盐水培养作为对照；B组：单核细胞用CoCl2溶液培养；C组：单核细胞用CrCl3溶液培养。分别在共培养12、24和48小时后，采用酶联免疫吸附测定（ELISA）法评估TNF-α的产生，采用比色法检测Caspase-3的激活情况，采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL）检测凋亡细胞。
B组和C组TNF-α浓度及Caspase-3表达均高于A组（P<0.05），且分别在24、48小时达到峰值水平。所有组均检测到TUNEL阳性细胞，细胞核固缩、染色加深，细胞体皱缩，细胞膜完整。B组、C组与A组凋亡率差异有统计学意义（P<0.05）。且Caspase-3的激活增加并与凋亡率呈正相关（r=0.765）。
Co2+和Cr3+离子可刺激人单核细胞释放TNF-α并诱导人单核细胞凋亡，导致假体周围骨溶解，且与Caspase-3的激活有关。
