IADR Abstract Archives
Dp7-c / Micro-26a Complex Regulates Osteogenesis by Regulating Osteogenic Immune Response
Objectives: Construct a new immunomodulatory peptide complex with microRNA ----DP7- C/miR-26a.Test the physical and biological properties,osteogenic properties and Immunoregulatory function of DP7- C/miR-26a complex.
Methods: We used agarose gel electrophoresis, nano particle size potentiometer, transfection experiment and CCK-8 Assay to test the physical and biological properties of DP7- C/miR-26a complex.Then, We cultured BMSCs and induced them into osteogenesis,add complex. their osteogenic properties were verified by RT-QPCR and Westerr Blot,modified alizarin red S staining.Then,LPS was added to RAW 264.7 cells to simulate the inflammatory states,add complex to Interfere with immune response. Used RT-qPCR(4h), ELISA(72h) and Griess method to test the Immunoregulatory function of DP7- C/miR-26a complex.
Results: After 3, 5 and 7 days of transfection, the results of alizarin red S and expression levels of ALP and BMP-2 in dp7-C group and complex group were significantly higher than 26a group (P<0.05); After 5 days, the expression of ALP in complex group significantly higher than DP7-C group (P<0.05); The result of Western blot shows that: The expression of ALP,OCN and BMP-2 protein in complex group were higher than 26a group and DP7-C group.Results of PCR and Elisa were illustrated together that expression levels and release of pro-inflammatory cytokines IL-1β, IL-6, TNF-α,iNOS and total nitric oxidein inDP7-C group and DP7-C/26a group were lower than those in control group (P<0.05). The result of elisa shows that The release of TGF-β in DP7-C group and DP7-C/26a group was significantly higher than that in control group (P< 0.01), and which in DP7-C/26a group was higher than in DP7-C group (P<0.05).
Conclusions: DP7-C/ mic-26a complex could promote osteogenic differentiation of BMSCs, and its effect is superior to mic-26A and DP7-C. DP7- C/miR-26a complex could reduce the expression and release of inflammatory factors in RAW264.7 in inflammatory state such as IL-1β,IL-6,TNF-α,NO, and increase the expression of TGF-β to regulate osteogenesis.
Methods: We used agarose gel electrophoresis, nano particle size potentiometer, transfection experiment and CCK-8 Assay to test the physical and biological properties of DP7- C/miR-26a complex.Then, We cultured BMSCs and induced them into osteogenesis,add complex. their osteogenic properties were verified by RT-QPCR and Westerr Blot,modified alizarin red S staining.Then,LPS was added to RAW 264.7 cells to simulate the inflammatory states,add complex to Interfere with immune response. Used RT-qPCR(4h), ELISA(72h) and Griess method to test the Immunoregulatory function of DP7- C/miR-26a complex.
Results: After 3, 5 and 7 days of transfection, the results of alizarin red S and expression levels of ALP and BMP-2 in dp7-C group and complex group were significantly higher than 26a group (P<0.05); After 5 days, the expression of ALP in complex group significantly higher than DP7-C group (P<0.05); The result of Western blot shows that: The expression of ALP,OCN and BMP-2 protein in complex group were higher than 26a group and DP7-C group.Results of PCR and Elisa were illustrated together that expression levels and release of pro-inflammatory cytokines IL-1β, IL-6, TNF-α,iNOS and total nitric oxidein inDP7-C group and DP7-C/26a group were lower than those in control group (P<0.05). The result of elisa shows that The release of TGF-β in DP7-C group and DP7-C/26a group was significantly higher than that in control group (P< 0.01), and which in DP7-C/26a group was higher than in DP7-C group (P<0.05).
Conclusions: DP7-C/ mic-26a complex could promote osteogenic differentiation of BMSCs, and its effect is superior to mic-26A and DP7-C. DP7- C/miR-26a complex could reduce the expression and release of inflammatory factors in RAW264.7 in inflammatory state such as IL-1β,IL-6,TNF-α,NO, and increase the expression of TGF-β to regulate osteogenesis.
IMAGES
3721186_File000000.jpg
3721186_File000000.jpg