Min Dai1, Xiaojun Yuan, Hongxian Fan, Min Cheng, Jiangbo Ai. 1. Department of Orthopaedics II, the First Affiliated Hospital, Nanchang University, Nanchang Jiangxi, 330006, P.R. China. daimin@medmail.com.cn
Abstract
OBJECTIVE: Lots of metal ions accumulation and over-expression of receptor activator of NF-kappaB ligand (RANKL) around the prosthesis could be found in revision of total hip arthroplasty. To investigate the relationship between metal ions and aseptic loosening by observing the effects of Co2+ and Cr3+ ions on the expression of RANKL and osteoprotegerin (OPG) from osteoblast. METHODS: Osteoblasts were cultured in vitro at the density of 1 x 10(5) cells/mL, and were divided into 2 groups according to different culture solutions. In control group, osteoblasts were cultured with normal medium without CoCl2 and CrCl3. In experimental group, osteoblasts were cultured with the medium including CoCl2 (10 mg/L) and CrCl3 (150 mg/L) solutions. The RT-PCR and ELISA methods were applied to detect the mRNA expression of RANKL and OPG and protein level at 24 and 48 hours after co-cultured, respectively. RESULTS: RT-PCR revealed that the mRNA expression of RANKL and OPG could be found in two groups at 24 and 48 hours after co-cultured, the expression was higher in the experimental group than in control group, especially the expression of RANKL, showing significant difference (P < 0.05). At 24 and 48 hours after co-cultured, the ratios of RANKL mRNA to OPG mRNA in the experimental group were 0.860 and 1.232, respectively, which were significantly higher than those in the control group (0.695 and 0.688, P < 0.05). ELISA revealed that the protein level of RANKL and OPG in experimental group were significantly higher than those in the control group (P < 0.05). CONCLUSION: Co2+ and Cr3+ can stimulate the mRNA expressions of RANKL, OPG and secretion of those protein from osteoblasts, especially increase of the RANKL, which promotes the formation and activation of osteoblasts and the generation of aseptic loosening.
OBJECTIVE: Lots of metal ions accumulation and over-expression of receptor activator of NF-kappaB ligand (RANKL) around the prosthesis could be found in revision of total hip arthroplasty. To investigate the relationship between metal ions and aseptic loosening by observing the effects of Co2+ and Cr3+ ions on the expression of RANKL and osteoprotegerin (OPG) from osteoblast. METHODS: Osteoblasts were cultured in vitro at the density of 1 x 10(5) cells/mL, and were divided into 2 groups according to different culture solutions. In control group, osteoblasts were cultured with normal medium without CoCl2 and CrCl3. In experimental group, osteoblasts were cultured with the medium including CoCl2 (10 mg/L) and CrCl3 (150 mg/L) solutions. The RT-PCR and ELISA methods were applied to detect the mRNA expression of RANKL and OPG and protein level at 24 and 48 hours after co-cultured, respectively. RESULTS: RT-PCR revealed that the mRNA expression of RANKL and OPG could be found in two groups at 24 and 48 hours after co-cultured, the expression was higher in the experimental group than in control group, especially the expression of RANKL, showing significant difference (P < 0.05). At 24 and 48 hours after co-cultured, the ratios of RANKL mRNA to OPG mRNA in the experimental group were 0.860 and 1.232, respectively, which were significantly higher than those in the control group (0.695 and 0.688, P < 0.05). ELISA revealed that the protein level of RANKL and OPG in experimental group were significantly higher than those in the control group (P < 0.05). CONCLUSION:Co2+ and Cr3+ can stimulate the mRNA expressions of RANKL, OPG and secretion of those protein from osteoblasts, especially increase of the RANKL, which promotes the formation and activation of osteoblasts and the generation of aseptic loosening.