Objective： To explore the influence of AD-VEGF-siRNA on the expression of vascular endothelial growth factor （VEGF） in neoplasm and blood serum. Methods： Transplantable model of human osteosarcoma was successfully...Objective： To explore the influence of AD-VEGF-siRNA on the expression of vascular endothelial growth factor （VEGF） in neoplasm and blood serum. Methods： Transplantable model of human osteosarcoma was successfully established by the way of subcutaneous injection of VEGF highly expressed human MG63 osteosarcoma cells. These mice were divided randomly into three groups： AD-VEGF-siRNA group, 15 mice; AD-EGFP group, 15 mice; PBS group, 15 mice. Three mice were additionally raised without any treatment. The drug was injected intratumorally 200 μL at each time, once a day. The total dose of virus was 2 × 10^9 pfu. Three osteosarcema-bearing mice of each group were sacrificed at 11th, 14th ,17th day after the implantation of MG63 cells. The expression of VEGF in implanted tumors and blood serum was detected by ELISA methods. Then the left mice were all sacrificed at the end of experiment （19th day）. The expression of VEGF in implanted tumors was detected by RT-PCR and immune histochemistry methods, and that in implanted tumors and blood serum was detected by ELISA methods. Results： （1） Tumors in mice could be seen at 5th day from the implantation of MG63 cells. （2） The expression of VEGF could be detected in all groups by RT-PCR and immune histochemistry, Which was much lower in the group receiving AD-VEGF-siRNA therapy than two control groups （P 〈 0.05）. （3） The expression of VEGF in blood serum of osteosarcoma-bearing mice was much higher than that of three healthy mice by ELISA （P 〈 0.05）. （4） The expression of VEGF in blood serum and neoplasm in AD-VEGF-siRNA group was much lower than that in two control groups （P 〈 0.05）. Conclusion： AD-VEGF-siRNA could effectively inhibited VEGF expression in vivo. This technology would bring some good references for our therapy of antiangiogenesis in osteosarcoma.展开更多
The balance of redox homeostasis is key to stem cell maintenance and differentiation.However,this balance is disrupted by the overproduced reactive oxygen species(ROS)in pathological conditions,which seriously impair ...The balance of redox homeostasis is key to stem cell maintenance and differentiation.However,this balance is disrupted by the overproduced reactive oxygen species(ROS)in pathological conditions,which seriously impair the therapeutic efficacy of stem cells.In the present study,highly dispersed fullerol nanocrystals with enhanced bioreactivity were incorporated into hydrogel microspheres using one-step innovative microfluidic technology to construct fullerol-hydrogel microfluidic spheres(FMSs)for in situ regulating the redox homeostasis of stem cells and promoting refractory bone healing.It was demonstrated that FMSs exhibited excellent antioxidant activity to quench both intracellular and extracellular ROS,sparing stem cells from oxidative stress damage.Furthermore,these could effectively promote the osteogenic differentiation of stem cells with the activation of FoxO1 signaling,indicating the intrinsically osteogenic property of FMSs.By injecting the stem cells-laden FMSs into rat calvarial defects,the formation of new bone was remarkably reinforced,which is a positive synergic effect from modulating the ROS microenvironment and enhancing the osteogenesis of stem cells.Collectively,the antioxidative FMSs,as injectable stem cell carriers,hold enormous promise for refractory bone healing,which can also be expanded to deliver a variety of other cells,targeting diseases that require in situ redox regulation.展开更多
基金a grant from the National Nature Sciences Foundation of China (No. 30271314)
文摘Objective： To explore the influence of AD-VEGF-siRNA on the expression of vascular endothelial growth factor （VEGF） in neoplasm and blood serum. Methods： Transplantable model of human osteosarcoma was successfully established by the way of subcutaneous injection of VEGF highly expressed human MG63 osteosarcoma cells. These mice were divided randomly into three groups： AD-VEGF-siRNA group, 15 mice; AD-EGFP group, 15 mice; PBS group, 15 mice. Three mice were additionally raised without any treatment. The drug was injected intratumorally 200 μL at each time, once a day. The total dose of virus was 2 × 10^9 pfu. Three osteosarcema-bearing mice of each group were sacrificed at 11th, 14th ,17th day after the implantation of MG63 cells. The expression of VEGF in implanted tumors and blood serum was detected by ELISA methods. Then the left mice were all sacrificed at the end of experiment （19th day）. The expression of VEGF in implanted tumors was detected by RT-PCR and immune histochemistry methods, and that in implanted tumors and blood serum was detected by ELISA methods. Results： （1） Tumors in mice could be seen at 5th day from the implantation of MG63 cells. （2） The expression of VEGF could be detected in all groups by RT-PCR and immune histochemistry, Which was much lower in the group receiving AD-VEGF-siRNA therapy than two control groups （P 〈 0.05）. （3） The expression of VEGF in blood serum of osteosarcoma-bearing mice was much higher than that of three healthy mice by ELISA （P 〈 0.05）. （4） The expression of VEGF in blood serum and neoplasm in AD-VEGF-siRNA group was much lower than that in two control groups （P 〈 0.05）. Conclusion： AD-VEGF-siRNA could effectively inhibited VEGF expression in vivo. This technology would bring some good references for our therapy of antiangiogenesis in osteosarcoma.
基金This work was supported by the National Key Research and Development Program of China(2020YFA0908200)National Natural Science Foundation of China(81772372 and 81930051)+2 种基金Shanghai Jiao Tong University“Medical and Research”Program(ZH2018ZDA04)Science and Technology Commission of Shanghai Municipality(18140901500,19440760400)Shanghai Municipal Health and Family Planning Commission(201840027).
文摘The balance of redox homeostasis is key to stem cell maintenance and differentiation.However,this balance is disrupted by the overproduced reactive oxygen species(ROS)in pathological conditions,which seriously impair the therapeutic efficacy of stem cells.In the present study,highly dispersed fullerol nanocrystals with enhanced bioreactivity were incorporated into hydrogel microspheres using one-step innovative microfluidic technology to construct fullerol-hydrogel microfluidic spheres(FMSs)for in situ regulating the redox homeostasis of stem cells and promoting refractory bone healing.It was demonstrated that FMSs exhibited excellent antioxidant activity to quench both intracellular and extracellular ROS,sparing stem cells from oxidative stress damage.Furthermore,these could effectively promote the osteogenic differentiation of stem cells with the activation of FoxO1 signaling,indicating the intrinsically osteogenic property of FMSs.By injecting the stem cells-laden FMSs into rat calvarial defects,the formation of new bone was remarkably reinforced,which is a positive synergic effect from modulating the ROS microenvironment and enhancing the osteogenesis of stem cells.Collectively,the antioxidative FMSs,as injectable stem cell carriers,hold enormous promise for refractory bone healing,which can also be expanded to deliver a variety of other cells,targeting diseases that require in situ redox regulation.
