| Research backgroundHeterotopic ossification(HO)is a histopathological abnormality of progressive bone formation that occurs primarily in non-bony soft tissues(muscles,tendons,ligaments).Traumatic heterotopic ossification(THO)is the most common type,which is commonly seen after burns and joint replacement.Severe THO can cause severe pain,limited joint movement or even disability,and reduce the quality of life of patients.Therefore,it is of great significance to explore new methods for the treatment of traumatic heterotopic ossification.Tendon Stem Cells(TSCs)are a subgroup of stem cells located in tendon integrity.In addition to having biological functions similar to those of mesenchymal stem cells such as autonomous replacement,proliferation and formation,they also have the ability of multidirectional differentiation,playing a decisive role in repairing tendon injury and maintaining environmental homeostasis in tendon tissues.Curcumin is a natural small molecular compound extracted from the root stems of the ginger,which can exert anti-infection,anti-inflammatory and antioxidant effects.Studies have shown that curcumin can induce cell migration,promote the repair of the dermis,nerve fibers and soft tissue injuries such as burns.The outside world believes that curcumin may have the ability to treat HO.Research Purpose1.To verify the identity of tendon stem cells as progenitor cells of ectopic ossification and the effect of curcumin on tendon stem cells.2.To study the relationship between ectopic ossification and inflammatory microenvironment in vitro and the effect of curcumin in inflammatory microenvironment.3.The effect of curcumin on heterotopic ossification of tendon in rats was verified in vivoResearch Methods1.Tendon stem cells were isolated from SD rats and cultured to identify their proliferation and multi-lineage differentiation ability.The optimal concentration of curcumin was screened,and the effects of curcumin were observed at the cellular and molecular protein levels.2.The inflammatory microenvironment was simulated in vitro,and the changes of NF-κB/P65 signaling pathway and osteogenic ability of TSCs were observed at the cellular and molecular protein levels,and the blocking effect of curcumin on the above processes was also observed.3.Micro 3D CT,HE,MASSON staining,immunofluorescence and histochemistry were used to further verify that inflammatory microenvironment activated NF-κB/P65 signaling pathway to stimulate the osteogenic differentiation of TSCs in vivo,and curcumin could reverse the above process.Results1.ARS and oil red O staining showed the formation of calcium nodules and oil droplets.The results of ALP staining and activity assay and ARS staining showed that calcium nodules and alkaline phosphatase were increased,and the m RNA and protein expressions of RUNX2,OCN and SOX9 were up-regulated after curcumin stimulation.2.The inflammatory microenvironment was simulated in vitro to observe the changes of NF-κB/P65 signaling pathway and the osteogenic ability of tendon stem cells at the cellular and molecular protein levels,as well as the inhibitory effect of curcumin on the above processes.3.The SD heterotopic ossification model of rats was successfully established.Micro 3D CT showed that the amount of heterotopic bone in the model group increased,and HE and MASSON staining showed a large number of inflammatory cell infiltration,multiple ossification areas,and perivascular proliferation.Immunofluorescence and immunohistochemistry showed that the expression levels of SOX9,RUNX2,OCN osteogenic genes and the core molecules of NF-κB/P65 signaling pathway P65 and SMAD1 were up-regulated in the model group.Conclusion1.TSCs have the ability of self-proliferation and multi-directional differentiation,and curcumin can promote their osteogenic differentiation.2.Inflammatory microenvironment plays an important role in the progression of HO by activating the NF-κB/P65 signaling pathway.3.Curcumin plays a role in the treatment of ectopic ossification by inhibiting the activation of NF-κB/P65 signaling pathway.Figure 12 Table 22 Reference 131... |
PDF Full Text Request