Magnetic resonance/fluorescence bimodal nanoprobe and preparation method thereof
A nano-probe and dual-mode technology, applied in the field of biomedical materials, can solve the problems of single nano-probe and difficulty in obtaining clear image results in imaging modes, and achieve clear image results, clear anatomical structure information, and good in vivo stability sexual effect
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[0051] The preparation method of the magnetic resonance / fluorescence dual-mode nanoprobe of the present invention specifically includes the following processes:
[0052] S1: Synthesis of SPIONs by hydrothermal method and pretreatment. details as follows:
[0053] 2mmol iron acetylacetonate, 10mmol 1,2-hexanediol, 6mmol oleic acid, 6mmol oleamide and 20mL benzyl ether were mixed into a round bottom flask, argon gas was introduced to remove the air in the reaction system and stirred. The mixture was heated to 200°C for 2h, then heated to 300°C for 1h under reflux. The resulting black mixture was cooled to room temperature, the product was precipitated with ethanol, centrifuged (10000 g, 10 min) to remove the solvent, and the product was dispersed in hexane to obtain a SPIONs solution. The obtained SPIONs solution was centrifuged at 3000 g to remove aggregates.
[0054] DSPE-PEG2000 AMINE molecules are coated on the surface of SPIONs through hydrophilic and hydrophobic interac...
Embodiment 1
[0079] This example illustrates the construction and evaluation of an in vitro BBB model.
[0080] Experimental method: An in vitro blood-brain barrier model was established to evaluate the permeability of bimodal nanoprobes. Both U87-MG cells and b.End 3 cells were cultured in high-glucose DMEM containing 10% fetal bovine serum and 1% penicillin / streptomycin at 37°C and 5% CO 2 cultivated under conditions. Apply a thin layer of collagen solution evenly on the inner side of the Transwell polymicroporous lipid membrane, place it on a clean bench for about 30 minutes, and use it after it dries naturally. Divide b.End 3 cells in 5×10 4 Cells / well were seeded in a 24-well Transwell upper chamber. Two days later, U87-MG cells were seeded at 1×10 5 Cells / well were seeded in the Transwell lower chamber and co-cultured for one week.
[0081] First, the constructed BBB model was tested and evaluated by leak test. Add 200 μL and 900 μL of culture medium to the upper and lower chamb...
Embodiment 2
[0089] This example evaluates the performance of the bimodal nanoprobe prepared by the method of the present invention to penetrate the BBB model in vitro.
[0090] Experimental method: After the in vitro BBB model was successfully constructed by the method in Example 1, the upper and lower chambers of the Transwell were gently washed twice with PBS, and PBS, Cy7-PEG-DSPE-SPIONs and two peptides / Cy7-PEG-DSPE were added to the upper chamber -SPIONs solution (two peptide / Cy7-PEG-DSPE-SPIONs solutions said in this embodiment are: THR / Cy7-PEG-DSPE-SPIONs and D THR / Cy7-PEG-DSPE-SPIONs. Similarly, the two peptide / Cy7-PEG-DSPE-SPIONs solutions mentioned in the following examples are also THR / Cy7-PEG-DSPE-SPIONs and D THR / Cy7-PEG-DSPE-SPIONs, not repeated), the concentration of each group is 10.0 μg / mL, a total of 200 μL; add 900 μL of PBS to the lower chamber, incubate for 60 min, remove 100 μL of the chamber buffer solution and put it into a PET black 96-well plate, A multifunctio...
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