| Objective:The terminal differentiation induction model of primary condylar chondrocytes was established to indcue the process of condylar mineralization to explore the relationship between FLRT3 and BMP signaling pathway;To further explore the influence of static magnetic field on terminal differentiation-related factors by FLRT3,the terminal differention induction model of primary condylar chondrocytes loaded with static magnetic field was established,which also provided a theoretical foundation for studying the influence mechanism of static magnetic field on maxillofacial development and reconstructionMethods:1.Found an in vitro mineralization differentiation model of condylar chondrocytes,observing calcified nodules by alizarin red staining,and detecting the expression and localization of BMP2 and Runx2 in condylar chondrocytes by immunofluorescence,and verifying The reliability of the terminal mineralization induced model.2.Detecting the expression and localization of FLRT3 in condylar chondrocytes by immunofluorescence.The expression trends of terminal mineralization proteins such as FLRT3,BMP2,Smad1/5/8,p-Smad1/5/8 and Runx2 were observed by Western blotting.The expression characteristics and intrinsic relationship of FLRT3 at different stages of chondrocyte differentiation were analyzed.3.The in vitro model of static magnetic field terminal mineralization was established by loading static magnetic field on mineralization-induced condylar chondrocytes.Western blotting was used to detect the effects of static magnetic field on mineralization differentiation induction for 3,8 and 15 days.FLRT3 and the protein mineralization differentiation were related.The expression changes of protein BMP2,Smadl/5/8,p-Smad1/5/8,Runx2,and the influence of static magnetic field on the terminal mineralization of chondrocytes.Results:1.The formation of calcified nodules was observed by alizarin red staining.Immunofluorescence staining showed BMP2 and Runx2 expression were enhanced,suggesting that the mineralization induced was treated,terminal mineralization occurs in chondrocytes,and the vitro of terminal differention induction model is reliable.2.After mineralization induction treatment,FLRT3 was found to be significantly enhanced in chondrocytes.The expression BMP2,Smad 1/5/8,p-Smad 1/5/8,Runx2 gradually decreased,while FLRT3 expression gradually increased from d3 to d8,but their protein expression was higher than the control group.3.The expression trends of FLRT3 and BMP2,Smad 1/5/8,Runx2 protein in the static magnetic field mineralization induction model were consistent with those in the second part,but their expression was higher than that in the control group.Conclusions:After treatment of condylar chondrocytes by terminaldiffetentiation-inducing fluid,the BMP signaling pathway closely related to terminal mineralization in chondrocytes was activated,it promoted the expression of related mineralization specific factors,and to some extent,simulated the terminal differentiation of chondrocytes.2.In the process of terminal mineralization induction,FLRT3 is closely related to BMP signaling pathway,suggesting that the two can synergistically regulate the mineralization and maturation of maxillofacial cartilage.3.Static magnetic fields affect the mineralization of chondrocytes through FLRT3 and BMP signaling pathways. |
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