Water-soluble ultrathin carbon nitride two-dimensional nanosheet as well as preparation method and application thereof

A two-dimensional nanometer and carbon nitride technology, which is applied in other methods of inserting foreign genetic materials, nanotechnology, nanotechnology, etc., can solve problems such as degradation and affecting the treatment effect, and achieve the effect of low toxicity

Pending Publication Date: 2022-03-04
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, once the exogenous genetic material enters the cell through endocytosis, it will be degraded by lysosomes, affecting the therapeutic effect

Method used

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  • Water-soluble ultrathin carbon nitride two-dimensional nanosheet as well as preparation method and application thereof
  • Water-soluble ultrathin carbon nitride two-dimensional nanosheet as well as preparation method and application thereof
  • Water-soluble ultrathin carbon nitride two-dimensional nanosheet as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A water-soluble ultra-thin carbon nitride two-dimensional nanosheet, using carbon nitride as a carrier, and further constructing the carbon nitride into a water-soluble ultra-thin two-dimensional nanosheet.

[0041] The preparation method of the water-soluble ultra-thin carbon nitride two-dimensional nanosheet of the above-mentioned present embodiment includes the following steps:

[0042] (1) Put 5 g of melamine in a crucible, cover the crucible, put it into a tube furnace for calcination under the protection of nitrogen, control the heating rate of the tube furnace at 5 °C / min, and keep it at 550 °C for 2 h. After the calcined product is cooled and ground, block carbon nitride is obtained.

[0043] (2) Add 1 g of block carbon nitride to 40 mL of a mixed acid solution of 98% concentrated sulfuric acid and 68% concentrated nitric acid and stir for 2 h. The ratio of concentrated sulfuric acid to concentrated nitric acid is 1:1. After the reaction, the white flocs were c...

Embodiment 2

[0048] Prepare a solution of 50 μg / mL ultra-thin carbon nitride nanosheets, incubate it with Hela cells for 12 h, and then use a 400 nm wavelength LED light source (450 mW / cm 2 ) were irradiated for different durations, and those without light were used as controls. Then replace the medium, incubate for another 12 h, and add 20 μL of MTT solution (5 mg / mL) to the cells. After 4 h of incubation, replace the medium with 200 µL DMSO. Cell viability was obtained by measuring the resulting solution in a Bio Tech Synergy H4 at a wavelength of 570 nm.

[0049] Figure 4 Is the carbon nitride nanosheet of the present invention under light conditions, the survival rate of Hela cells; by Figure 4 It can be seen that the survival rate of Hela cells is getting lower and lower as the time increases under the condition of light of carbon nitride nanosheets. The carbon nitride nanosheets prepared by the invention can promote the apoptosis of cancer cells under light conditions. It can be...

Embodiment 3

[0051] Different concentrations of carbon nitride nanosheets and pUC18 plasmid DNA (9 μg / mL) were placed in a 37 ºC water bath, incubated for 1 h, and then DNA agarose gel retardation experiments were performed to obtain the gel retardation of different concentrations of compounds on pUC18 DNA. stagnant results.

[0052] Figure 5 It is the result of the agarose gel retardation experiment of carbon nitride nanosheets of the present invention to pUC18 DNA; Figure 5 The value marked in the middle and above is the test concentration (μg / mL); Figure 5 It can be seen that the carbon nitride nanosheets prepared in the present invention can completely block the migration of DNA in agarose at a lower concentration. It can be concluded from Example 3 that the water-soluble ultra-thin carbon nitride nanosheets prepared by the present invention can effectively condense DNA to form nanoparticles, and can be used as a non-viral gene carrier.

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Abstract

The invention discloses a preparation method and application of a water-soluble ultrathin carbon nitride two-dimensional nanosheet, and the preparation method comprises the following steps: calcining a nitrogen-containing precursor at 550 DEG C to obtain initial blocky carbon nitride; stripping the initial blocky carbon nitride in a mixed acid solution of concentrated sulfuric acid and concentrated nitric acid to obtain porous carbon nitride; performing long-time ultrasonic stripping reaction on the porous carbon nitride to obtain a final product. The method is simple in preparation process, low in cost and good in repeatability. The nanosheet provided by the invention has a good fluorescence imaging function and can be used for dyeing cells; the nanosheet provided by the invention can generate a large amount of active oxygen under an illumination condition, so that the function of photodynamic therapy is realized; the nanosheet provided by the invention can be used as a photoresponsive non-viral gene vector, so that the delivery of DNA is promoted, and cell nucleuses can be targeted. The nanosheet provided by the invention can load a P53 gene and can realize gene and photodynamic combined therapy on tumors.

Description

technical field [0001] The invention relates to a non-viral gene carrier, in particular to a water-soluble ultra-thin carbon nitride two-dimensional nanosheet, a preparation method and application thereof. Background technique [0002] Gene therapy refers to the delivery of nucleic acids with therapeutic functions (such as DNA, mRNA and siRNA) to target cells to regulate protein expression and further achieve the effect of disease treatment. As a new type of treatment, gene therapy has attracted extensive attention from scientific researchers. However, once the exogenous genetic material enters the cell through endocytosis, it will be degraded by lysosomes and affect the therapeutic effect. Therefore, constructing a safe and efficient gene delivery system is a decisive factor for the success of gene therapy. [0003] As a new type of graphene-like material, carbon nitride has gradually aroused great interest in the field of biomedicine due to its excellent physical and c...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K41/00A61P35/00B82Y5/00C12N15/87
CPCA61K48/0033A61K41/0057A61P35/00B82Y5/00C12N15/87
Inventor 刘名轩张小玲
Owner NANTONG UNIVERSITY
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