A dual-modal nanoprobe targeting brain glioblastoma and its preparation method
A glioblastoma and nanoprobe technology, applied in the field of biomedical materials, can solve problems such as difficulty in obtaining clear image results, single imaging mode of nanoprobes, etc., and achieve the effect of clear image results
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[0051] The preparation method of the bimodal nanoprobe targeting brain glioblastoma of the present invention specifically includes the following process:
[0052] S1: Synthesis of SPIONs by hydrothermal method and pretreatment. details as follows:
[0053] Mix 2mmol iron acetylacetonate, 10mmol 1,2-hexanediol, 6mmol oleic acid, 6mmol oleamide and 20mL benzyl ether into a round bottom flask, blow in argon to remove the air in the reaction system and stir. 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, the solvent was removed by centrifugation (10000 g, 10 min), and the product was dispersed in hexane to obtain a SPIONs solution. The obtained SPIONs solution was centrifuged at 3000g to remove aggregates.
[0054] DSPE-PEG2000AMINE molecules are coated on the surface of SPIONs through hydrophilic-hydrophobic interaction, and the PEGylated ma...
Embodiment 1
[0077] This example illustrates the construction and evaluation of an in vitro BBB model.
[0078] 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 wait for it to dry naturally before using it. Divide b.End 3 cells in 5×10 4 The density of cells / well was seeded in the upper chamber of 24-well Transwell. Two days later, U87-MG cells were seeded at 1×10 5 The density of cells / well was seeded in the lower chamber of Transwell, and the co-culture was continued for one week.
[0079] First, the constructed BBB model was tested and evaluated by leak test. A...
Embodiment 2
[0087] This example evaluates the performance of bimodal nanoprobes penetrating the BBB model in vitro.
[0088] 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 are: ANG / Cy7-PEG-DSPE-SPIONs and D ANG / Cy7-PEG-DSPE-SPIONs. Similarly, the two peptide / Cy7-PEG-DSPE-SPIONs solutions mentioned in the following examples are also ANG / Cy7-PEG-DSPE-SPIONs and D ANG / 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 multifunctional microplate reader measures the fluorescence intensity of the lower chamb...
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