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BACKGROUND: In recent years, the use of circular RNA (circRNA) to regulate stem cell differentiation has become a new research focus. Therefore, loading circRNA on bone marrow mesenchymal stem cells to study their osteogenic properties is of great significance for the development of new materials to repair bone defect. OBJECTIVE: To investigate the osteogenic effect of circRNA mmu_circ_0001775 knockdown on bone marrow mesenchymal stem cells. METHODS: Bioinformatics analysis tools speculated that the upstream circRNA most closely related to miR-335-5p was mmu_circ_0001775. Lentivirus and negative control virus knockdown mmu_circ_0001775 were transfected into bone marrow mesenchymal stem cells. qRT-PCR was used to detect the expression levels of Runt-related transcription factor 2, osteocalcin, miR-335-5p, and mmu_circ_0001775 genes at 7 and 14 days of osteogenic induction, respectively, to verify their osteogenic effects. Western blot assay was used to detect the expression levels of bone morphogenetic protein 2 and osteopontin. RESULTS AND CONCLUSION: The expression levels of Runt-related transcription factor 2 and osteocalcin at each time point in LV-mmu_circ_0001775-BMMSCs group were higher than those in LV-BMMSCs group (P < 0.01). The relative expression levels of bone morphogenetic protein 2 and osteopontin proteins were significantly higher in the LV-mmu_circ_0001775-BMMSCs group than those of LV-BMMSCs group (P < 0.05). The results showed that cirCRNA mmu_ circ_0001775 knockdown could enhance the osteogenic ability of bone marrow mesenchymal stem cells. [ABSTRACT FROM AUTHOR]

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BACKGROUND: In recent years, the use of circular RNA (circRNA) to regulate stem cell differentiation has become a new research focus. Therefore, loading circRNA on bone marrow mesenchymal stem cells to study their osteogenic properties is of great significance for the development of new materials to repair bone defect. OBJECTIVE: To investigate the osteogenic effect of circRNA mmu_circ_0001775 knockdown on bone marrow mesenchymal stem cells. METHODS: Bioinformatics analysis tools speculated that the upstream circRNA most closely related to miR-335-5p was mmu_circ_0001775. Lentivirus and negative control virus knockdown mmu_circ_0001775 were transfected into bone marrow mesenchymal stem cells. qRT-PCR was used to detect the expression levels of Runt-related transcription factor 2, osteocalcin, miR-335-5p, and mmu_circ_0001775 genes at 7 and 14 days of osteogenic induction, respectively, to verify their osteogenic effects. Western blot assay was used to detect the expression levels of bone morphogenetic protein 2 and osteopontin. RESULTS AND CONCLUSION: The expression levels of Runt-related transcription factor 2 and osteocalcin at each time point in LV-mmu_circ_0001775-BMMSCs group were higher than those in LV-BMMSCs group (P < 0.01). The relative expression levels of bone morphogenetic protein 2 and osteopontin proteins were significantly higher in the LV-mmu_circ_0001775-BMMSCs group than those of LV-BMMSCs group (P < 0.05). The results showed that cirCRNA mmu_ circ_0001775 knockdown could enhance the osteogenic ability of bone marrow mesenchymal stem cells. [ABSTRACT FROM AUTHOR]

BACKGROUND: Foreign body reactions will lead to the failure of material implantation. In recent years, research on immunity has gradually become a hot spot. It is of great significance to select scaffold materials with good immune properties for the development of bone defect repair. OBJECTIVE: To investigate the effects of calcinated antler cancellous bone and untreated antler bone on the biological behavior of bone marrow-derived M1 macrophages. METHODS: Calcinated antler cancellous bone was prepared by physical and chemical methods. The physical and chemical properties of calcinated antler cancellous bone and untreated antler bone were detected by X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. The extracts of calcinated antler cancellous bone and untreated antler bone were prepared and the contents of Ca, P and Mg in the extracts were determined. Bone marrowderived macrophages were cultured in vitro and polarized into M1 macrophages under lipopolysaccharide and interferon γ stimulation. These cells were then co-cultured with different extracts in calcinated antler cancellous bone and untreated antler bone groups, while co-cultured with an IMDM medium in control group, followed by cell counting kit-8 detection of cell survival, flow cytometry detection of M1 markers, and qPCR detection of macrophage-related genes. RESULTS AND CONCLUSION: (1) The contents of Ca, P and Mg were higher in the calcinated antler cancellous bone group than the untreated antler bone group. (2) The main functional groups in these two groups included OH-, PO4 3-, and CO3 2 -. (3) There was no significant difference in cell survival rate among the three groups for 1, 3, and 7 days of co-culture. (4) The expression rate of CD16/32 was lower in the calcinated antler cancellous bone group than the control group after 1-day co-culture. (5) The expression levels of pro-inflammatory cytokines, tumor necrosis factor-α and interleukin-6 were lower in the calcinated antler cancellous bone group than the control group after 1 and 3 days of co-culture, while there was no significant difference between the untreated antler bone group and the control group. The expression of CD206, a main marker of M2 macrophages, was significantly higher in the calcinated antler cancellous bone group than the control group at 3 days of co-culture, while there was no significant difference between the untreated antler bone group and the control group at 1 and 3 days of co-culture. (6) To conclude, calcinated antler cancellous bone powder extract can reduce the expression of pro-inflammatory cytokines in M1 macrophages and promote their polarization to M2 macrophages [ABSTRACT FROM AUTHOR]