【摘要】 目的观察人工关节磨损产物Co2+、Cr3+刺激下人单核细胞生物反应过程中TNF-α的表达,以及人单核细胞凋亡与Caspase-3的变化,探讨Co2+、Cr3+引发假体周围骨溶解的作用机制。方法将CoCl2粉末与CrCl3粉末溶于无菌注射用水,分别配制成浓度为10mg/L和500mg/L的CoCl2溶液和CrCl3溶液。从健康志愿者外周血分离培养单核细胞,调整浓度为4×106个/培养皿,根据培养液不同,分为3组。A组生理盐水作为对照,B组CoCl2溶液,C组CrCl3溶液。于培养12、24及48h后吸取细胞上清液,ELISA法检测TNF-α含量,TUNEL检测细胞凋亡情况,比色法测定Caspase-3活性。结果各时间点B、C组TNF-α含量及Caspase-3活性均高于A组(P<0.05),分别于24h及48h达峰值。各时间点3组均存在TUNEL染色阳性细胞,细胞核固缩深染,细胞体积皱缩,胞膜完整。各时间点B、C组细胞凋亡率与A组比较,差异有统计学意义(P<0.05)。Caspase-3活性与细胞凋亡率成正相关(r=0.765)。结论Co2+、Cr3+均能刺激人单核细胞释放TNF-α并诱导其凋亡;细胞凋亡参与了假体周围骨溶解的病理过程,且可能与Casepase-3的激活有关。更多还原

【Abstract】 Objective To observe the human mononuclear cell releasing TNF-α and the activation of Caspase-3 during apoptosis after stimulated by Co2+ and Cr3+,to discuss the mechanism of arti cial joint wear production metal ion on aseptic loosening. Methods 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,4×106/culture dish. According to the di erence 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 CoCl2 solution; group C: mononuclear cell was cultured with CrCl3 solution. The production of TNF-α 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. Results The concentration of TNF-α and theffexpression 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,respetively. 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 signi cant di erences 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). Conclusion Co2+ and Cr3+ ions can stimulate human mononuclear cell to release TNF-α and induce human mononuclear cell apoptosis,which result in periprosethetic osteolysis and related to activation of Caspase-3.更多还原