Window Chamber Observation And HIFU Synergistic Effect Of Targeting CD133 Fluorescent Fluorocarbon Nanomicelle In Nude Mice

Part ? Preparation,Characterization and Ultrasound Imaging of CD133 Silicone Fluorocarbon Fluorescent Nano-micellesOBJECTIVE:To prepare CD133-targeted nanoparticles(CD133NPs)targeting CD133 and evaluate their physicochemical properties and safetyMETHODS:Targeted CD133NPs and IgG-targeted nanoparticles(IgGNPs)were prepared by atomic transfer polymerization(ATRP)and other mature polymer polymerization techniques.The physicochemical properties such as morphology and size were detected and detected by CCK-8 kit.Micellar inhibition of cell proliferationRESULTS:CD133NPs and IgGNPs were successfully prepared,and the appearance was spherical.The particle size was about 100 nm by transmission electron microscopy.The zeta potential and thermal specific gravity analysis confirmed the successful attachment of silane and antibody.The cell proliferation inhibition experiment verified that the 5mg/ml micelles were on the cells.The effect of proliferation is smallConclusion:Targeting CD133 organosilicon fluorocarbon fluorescent nanomicelles are nano-sized particles,which can be deformed after HIFU excitation,and have better in vitro two-dimensional ultrasound imaging ability.Part ? In vitro targeting experiments targeting CD 133 organosilicon fluorocarbon fluorescent nanomicellesObjective:To investigate the in vitro targeting ability of CD133NPsMETHODS:CD133-positive pancreatic cancer stem cells from BXPC-3 pancreatic cancer cells were sorted by immunomagnetic beads sorting.The expression of CD 133 in pancreatic cancer stem cells was determined.The expression of stem cell marker protein OCT-4 was verified by immunoblotting.The pancreatic cancer stem cells were co-cultured with CD133NPs and IgGNPs for 24 hours,and their in vitro targeting ability was evaluated.RESULTS:Immunomagnetic beads sorting technique successfully sorted CD133+pancreatic cancer cells.After culture,CD133+pancreatic cancer stem cells were significantly different from normal cell morphology and growth pattern.Immunoblot analysis showed that stem cell marker protein OCT-4 was positive for CD133.The expression of pancreatic cancer stem cells was significantly higher than that of the negative cells(P<0.01).After incubation with CD 133NPs and IgGNPs,pancreatic cancer stem cells showed higher binding to CD133NPsConclusion:The CD133+pancreatic cancer cells sorted by immunomagnetic beads technique have stem cell characteristics,and the prepared CD133NPs micelles have better in vitro active targeting abilityPart ? Imaging experiment of targeting CD 133 organosilicon fluorocarbon fluorescent nanomicelles in the back window model of nude miceOBJECTIVE:To investigate the in vivo targeting ability of CD133NPs in the spinal dorsal window model of nude miceMETHODS:The window model of spinal dorsal pancreatic cancer in nude mice was made and improved.The tumor and blood vessels in the window model of spinal dorsal pancreatic cancer were observed by two-photon fluorescence microscope after injection of CD133NPs into the tail vein.The mice were injected with CD133NPs and IgGNPs 4h and 8h after the tail vein.Ultrasound imaging was performed at 12h,24h and 48h to observe the echogenic changes in the subcutaneous tumor of pancreatic cancer.RESULTS:The two-photon fluorescence microscope was used to observe the window model of the dorsal pancreatic cancer in nude mice.The micelles in the window model were green.The green fluorescence of the micelles in the tumor window was quantitatively analyzed at Oh,24h and 48h after CD133NPs injection.The fluorescence intensity was the brightest,and the fluorescence intensity at 48h was weaker than that at 24h.Ultrasound imaging showed that the internal echo of the subcutaneous xenografts in the CD133NPs group was increased,and the increase was most obvious at 24h after injection.Conclusion:The window model of the dorsal pancreatic cancer in nude mice can observe the targeting of CD133NPs in vivo,and lay the experimental foundation for HIFU combined with micellar ablation of pancreatic cancer in nude micePart ? HIFU synergistic experiment targeting CD133 organosilicon fluorocarbon fluorescent nanomicelleOBJECTIVE:To investigate the ability of CD133NPs to enhance the effect of HIFU on subcutaneous transplantation of pancreatic cancer in nude miceMETHODS:Polyacrylamide gel containing different concentrations of organosilicon fluorocarbon nanomicelles was irradiated by HIFU to observe the size of the ablation lesions.CD133NPs,IgGNPs,saline group and control group were injected into the tail vein respectively,24 hours after injection.HIFU was used to irradiate subcutaneous xenografts of pancreatic cancer in nude mice,and the echo changes in the tumor were observed.After anatomy,HE staining was performed to observe the coagulative necrosis of the lesions and CD31 immunohistochemical analysis of the internal vascular damage RESULTS:After HIFU irradiation of micelle-containing polyacrylamide gel,the ablation area was larger than that of the micelle group,and the ablation area was positively correlated with the micelle concentration.HIFU ablation of pancreatic cancer subcutaneous tumor in nude mice,ultrasound showed injection The internal echo of CD133NPs and IgGNPs micelles increased,and the gray difference before irradiation was 121.35�1.89 and 68.32�0.23 respectively,P<0.05.Tumor volume reduction was most pronounced after HIFU ablation in the CD133NPs group.HE staining and CD31 immunohistochemical analysis showed that the tumor necrosis was the highest in the HIFU group after injection of CD133NPs,and the degree of vascular damage was the most obvious.Conclusion:CD133NPs have a good synergistic effect on HIFU treatment of pancreatic cancer,and provide a good integrated research platform for diagnosis and treatment of HIFU for pancreatic cancer.