Effects Of Gsk-3? On The Polarization Of Macrophages During Orthodontic Tooth Movement

Orthodontic tooth movement is a very complex process,including soft tissue remodeling,bone remodeling,changes in blood and cytokines.One of the most important is the remodeling of bone tissue,and bone remodeling is closely related to inflammatory response.The bone remodeling includes the pressure side,inflammation,promote osteoclast activity and bone breaking which is the premise of orthodontic tooth movement;the tension side due to tension,promote osteoblast activity and bone formation.As an important innate immune cell,macrophages plays an important role in inflammatory reaction,and the article point out that it has an important role in bone remodeling,as osteoclast precursor cells can regulate osteoclast activity.Macrophages were divided into M1 and M2 according to their functions.These two kinds of cells can be transformed into each other in a variety of physical and chemical environments.When the body inflammation,the recruitment of macrophages to sites of inflammation,inflammatory cytokines promote macrophage to M1 polarization,when inflammation is late,the physical and chemical changes in the environment,M1 macrophages into M2 macrophages and promote inflammation.Yanheng Zhou et al found that the ratio of M1 to M2 could affect the tooth movement and root resorption during tooth movement.Inhibition of M1 type macrophages can inhibit tooth movement,while in vivo injection of type M1 macrophages can alleviate this inhibition.It can be found that M1 and M2 type macrophages play an important role in orthodontic tooth movement.Gsk-3? is a serine /threonine protein kinase that plays an important role in the Wnt and Rankl pathways and can regulate bone remodeling and inflammation.The experimental results showed that the bone of GSK-3? gene deficient mice was increased,and the bone was significantly increased in the process of maxillary expansion in rats injected with Licl.The inhibition of GSK-3? could inhibit the root resorption during orthodontic tooth movement.In the orthodontic tooth movement,the GSK-3? and the macrophage polarization related factors were significantly changed.Therefore,we propose that GSK-3 beta plays an important role in the polarization of macrophages during orthodontic tooth movement.The experiment is divided into three parts:1.The model of orthodontic tooth movement in miceObjective:To explore the effective method to establish the model of orthodontic tooth movement in mice.Method:Methods 30 male C57 mice were randomly divided into0 d,1d,3d,5d,7d.Except the 0d group between each mouse maxillary right incisor and first premolar for orthodontic spring.During the process of power supply,the fall off rate is measured periodically.Each group was sacrificed at the corresponding time,and then the teeth were moved to observe the movement of the teeth after scanning and microCT.Bone regeneration was observed by HE staining and immunofluorescence staining after decalcification.Result: The results of microCT showed that the teeth moved significantly,and HE staining showed that there was bone remodeling on the pressure side.Immunofluorescence staining of trap showed that there was a broken bone on the pressure side and the pressure side of the periodontal ligament narrowed.Conclusions:Mouse model of tooth movement made successfully.2.To explore the impact of orthodontic force in the process of GSK-3 beta on macrophagepolarization in vivo.Objective:To explore the impact of orthodontic force in the process of GSK-3 beta on macrophage polarization in vivo.Method:30 wild C57 mice and 20 GSK-3 knockout C57 mice were selected,respectively.20 wild-type and 20 knockout type,after 3D 5D,7d orthodontic force,and 14d(n = 5)were sacrificed,then fixed jaw scanning MicroCT measurement of tooth movement distance,frozen sections immunofluorescence staining to observe osteoclasts,macrophages and M1 macrophages.Wild type 10 were intraperitoneally injected with Licl(the next day 100ng/ml)augmented with 3D,7d was killed after scanning MicroCT.Result: Comparing the gene knockout with wild type group injected Licl group,there is no significant difference between the distance of tooth movement;compared with the wild type group at the same time,the distance of GSK-3gene knockout mice tooth movement reduced,pressure side of osteoclasts reduced,macrophages and M1 macrophages decreased,which was statistically significant.Conclusions:GSK-3? gene knockout mice comparing with wild-type mice,the distance of orthodontic tooth movement is reduced,the number of macrophages and M1 macrophages also decreased,suggesting that GSK-3 beta may affect the macrophage polarization effect of orthodontic tooth movement.3.Effect of GSK-3? on macrophage polarization in vitroObjective: To explore the effect of GSK-3? on macrophage polarization in vitro.Method: Peritoneal macrophages were extracted from gsk-3? gene knockout mice and wild-type mice,cell culturing to observe the positive rate of immunofluorescence staining after one week;peritoneal macrophages were extracted from gsk-3? gene knockout mice and wild-type mice,after a week of training,LPS(100ng/ml)after 2d induction,testing flow cytometry(iNOS)ratio of M1 macrophages.Result: The positive rate of macrophage extraction could reach more than 90%.The results of flow cytometry showed that the positive rate of M1 macrophage in GSK-3 knockout mice was significantly lower than that of wild type.Conclusions:GSK-3? can affect the polarization of macrophages.