| With the increasing of population aging and the expected life, artificial joint replacement has been widely carried out and applied in the surgical field. According to the latest statistics, only in America, there are more than 1 million hip and knee joint replacement operations every year. Artificial joint replacement is the effective method recognized to treat advanced bone joint diseases. However, new difficulties and challenged have been proposed clinically, i.e. how to relieve the negative effect and adverse influence of brought by the osteolysis surrounding the prosthesis. The aseptic loosening followed by the osteolysis surrounding artificial joint prosthesis is one of the most important reasons for joint overhaul after artificial joint replacement. The UHMWPE particles generated in the traditional metal-polyethylene fraction interface induce the inflammatory activation of macrophages in the interference membrane surrounding the prosthesis, and a lot of inflammatory factors cause the activation of bone cells to trigger osteolysis surrounding the prosthesis. In order to extend the service life of prosthesis and avoid the high cost of overhaul operation, we are devoted to discussing a safe and effective clinical drug to prevent and treat the aseptic loosening followed byosteolysis surrounding the prosthesis.At present, non-steroid anti-inflammatory drug and bisphosphonate drugs are used to prevent and treat the aseptic loosening of osteolysis after artificial joint replacement, but these drugs have a significant adverse effect and cannot be used for a long time generally. The latest pharmacological research has found that in addition to malaria resistance, dihydroartemisinin also has the role of efficient inflammation resistance. But there has been no literature report on the use of dihydroartemisinin in prevention and treatment of aseptic loosening of osteolysis after artificial joint replacement.PurposeIn the in vivo experiment, it is to research the influence of dihydroartemisinin on endogenus inflammatory factors release of mice macrophage system RAW264.7 induced by ultra-high molecular weight polyethylene particles. Under the condition of in vitro experiment, the influence of dihydroartemisinin on the mice skull osteolysis induced by ultra-high molecular weight polyethylene particles is discussed.MethodExperiment I: The effects of Dihydroartemisinin on Macrophage-derived inflammatory factor production induced by UHMWPE particlesUHMWPE particles were used to establish inflammatory factors production model in RAW264.7 cell and it was treated with dihydroartemisinin on different concentrations. The content of TNF-α, IL-1, IL-6 and IL-10 in the supernatant of cell culture medium was detected by ELISA. MTT assay was adopted to detect the cytotoxicity of dihydroartemisinin.Experiment II: Effect of dihydroartemisinin on mice skull osteolysis induced by UHMWPE particlesThe model of mice skull osteolysis induced by UHMWPE particles is established; different concentrations of dihydroartemisinin are added, after culture of skull tissue for 24 h, the supernate was collected, and the contents of TNF-α, IL-1β, IL-6 and IL-10 in the cell culture supernate were detected by enzyme-linked immuno sorbent assay(ELISA).Through Micro-CT scanning, precise 3D reconstruction was performed to the skull and various parameters were analyzed, including mineral concentration and bone volume fraction. The skull specimens were subject to histopathological staining, and htoxylin eosin staining and tartrate-resistant acid phosphatase staining were used. Osteoporosis Measure Analysis System and inverted phase contrast microscope and photographic system were adopted to observe H&E staining slice, and the osteolysis area within the targeted area was calculated. TRAP staining slice observation as performed, and osteoclasts were counted in the targeted area.ResultExperiment IDihydroartemisinin could significantly inhibit the secretion of TNF-α, IL-1 and IL-6 in RAW246.7 cells induced by UHMWPE particles while promote the content of anti-inflammatory factor IL-10 in a dose-dependent manner(P<0.05)。Experiment II1. The result of ensyme-linked immunosorbent assay showed that dihydroartemisinin could significantly inhibit the expression of proinflammatory cytokines TNF-α, IL-1 and IL-6 of mice skull osteolysis model induced by UHMWPE particles, and significantly promote the release of anti-inflammatory cytokine IL-10, with effective of dosage dependence(P<0.05)。2. It was found that there was obvious osteolysis in the skull coronal suture bregma area in the positive control group, low-dosage group and high-dosage drug group through Mirco-CT scanning region of interest(ROI) 2Dsection, the positive control group has the maximum area of bone absorption, the drug-use group has a significantly reduced area, and the bone destruction in the high-dosage group was less. Mirco-CT parameters BMD and BV/TV had the same change trend. Both parameters were of the lowest value in the positive control group, indicating that the explicit destruction of osteolysis was serious. The value in the drug-use group increased, and the value in the high-dosage drug group rose more significantly than in the low-dosage drug group, indicating that the dosage group of dihydroartemisinin effectively inhibited the occurring of osteolysis(P<0.05)。3. H&E staining under microscope was visible. In relative to the blank control group, the positive control group had significant periosteal thickening, and a lot of inflammatory cells infiltrated and deposited the proliferation of fibrous tissue of UHMWPE particles; the continuous bone destruction in the mice skull tissue was significant, and the osteolysis area increased significantly. Different dosages of dihydroartemisinin drug use groups were observed, it was found that compared with the low-dosage group, the skull inflammation reaction in the high-dosage group reduced significantly, and the infiltration of inflammatory cells and continuous bone destruction reduced significantly(P<0.05)。Osteoporosis Measure Analysis System and inverted phase contrast microscope and photographic system were used to observe H&E staining slices, and the osteolysis area in the targeted area also reflected such trend. Through TRAP staining slice observation under the microscope, it was found that the irregular amaranth or bright red particles of multinuclear giant cells were presented in the cytoplasm of targeted area, with bone resorption of different areas in the surrounding areas. The counting of osteoclasts was visible, the positive control group had increasing significant activation of osteoclasts, the number of osteoclasts in the drug use group reduced greatly, and the inhabitation role of high-dosage group was significant than the low-dosage group(P<0.05) ConclusionDihydroartemisinin has significant anti-inflammation effect, which can significantly inhibit the macrophage inflammatory response induced by UHMWPE particles in vivo experiment, and in the in vitro experiment, it can also inhibit the occurring of generation and osteolysis of mice skull osteoclasts induced by UHMWPE grains, and it has a potential effect in the prevention of drug treatment of aseptic loosening after artificial joint replacement. |
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