Nanoparticle, preparation process and uses thereof
a nanoparticle and nanoparticle technology, applied in the field of nanoparticles, can solve the problems of poor biocompatibility of conventional nanoparticles, poor stability, and accumulation in the patient's central nervous system and skin, and achieve the effects of no damage to normal tissues, good biocompatibility and stability, and alleviation of tumor hypoxia
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example 1
Preparation and Characteristics of Nanoparticles of Present Invention
[0080]FIG. 1A is a process flow diagram showing the preparation of the nanoparticles of the present invention. First, ascorbic acid (Sigma Aldrich) was used to reduce potassium permanganate (KMnO4) (fisher scientific, Waltham, Mass.) to synthesize a core containing manganese dioxide, while a negatively charged lipid 1,2-dioleoyl-sn-glycero-3-phosphate (DOPA) (Avanti Polar Lipids, Alabaster) was used to coat the core. Briefly, two kinds of microemulsions (3 mL each), one contained DOPA lipid and KMnO4, another contained ascorbic acid, were prepared separately. To prepare the Mn7+-loaded microemulsions, 74 μL of 35 mM DOPA and 40 μL of 300 mM KMnO4 were dropped to the oil phase of cyclohexane and Igepal-520 (7:3, v / v). To prepare the ascorbic acid buffer-loaded microemulsions, 100 μL of 250 mM ascorbic acid and 900 μL of 1 M NaOH were mixed as ten times ascorbic acid buffer acting as reducing agent, and 40 μL of asco...
example 2
pH-Dependent Reactivity, Drug Release Profiles and Oxygen Generation Effect of Nanoparticles of Present Invention
[0091]Oxidative stress caused by tumor growth can significantly increase the concentration of H2O2 in the tumor microenvironment. In order to utilize the endogenous H2O2 produced by cancer cells and generate O2 in situ, the MnO2 cores must be delivered to the tumor region.
[0092]The reactivity of the PMD NPs toward H2O2 was then investigated by observing H2O2 quenching and O2 generation. Herein, a PeroXOquant assay kit (Pierce, USA) was used to study if PMD NPs would be able to quench H2O2 with pH-dependent reactivity. For the quenching experiment, PMD NPs (750 μM) or vehicle NPs (without MnO2 cores) were placed in Na2HPO4—NaH2PO4 buffer at pH 5.5 or 7.4, and H2O2 (300 μM) was added to initiate the reaction. The residual amount of H2O2 was determined over time using the PeroXOquant assay kit. The result is shown in FIG. 3A.
[0093]FIG. 3A is a schematic diagram showing the q...
example 3
In Vitro Cellular Uptake and Efficacy of SP94-Conjugated NPs
[0096]The NPs modified with tumor targeting peptides SP94 are the suitable carrier for the MnO2 cores. The cellular uptake of targeted or non-targeted PMD NPs in HCA-1 murine liver cancer cells (provided by Dr. Dan Duda) and Hep3B human liver cancer cells (purchased from the American Type Culture Collection (ATCC)) was measured.
[0097]Coumarin 6 (C6), a hydrophobic small molecule, was used as a fluorescent tracer in the PLGA-based NPs, formulated with a final weight ratio of C6 to PLGA as 1 / 150. HCA-1 or Hep3B cells (1×104 cells / mL) were seeded in the 12-well plates (Costar, Ill., USA) and incubated for 12 hours. The cells were then treated with C6-containing SP94 PMD (i.e., SP94 PLGA-based manganese dioxide (PMD)-C6 NPs) or C6-containing PMD NPs (i.e., PMD-C6 NPs), at 37° C. for 4 hours. After removing the medium in each well, the cells were washed with PBS and fixed with 4% paraformaldehyde for 10 minutes. After aspiration...
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