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Application of nitrogen-doped carbon quantum dot delivery system in cartilage tissue

A carbon quantum dot and delivery system technology, applied in the field of biomedicine, can solve the problems of untraceability, low transfection efficiency, and high toxicity and side effects, and achieve clear and stable fluorescent signals, simple transfection process, and high biocompatibility Effect

Pending Publication Date: 2021-12-31
THE SECOND HOSPITAL OF SHANXI MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The shortcomings of the existing delivery system are: poor biocompatibility, high toxicity and side effects; single effect; poor targeting; low transfection efficiency; large delivery system cannot pass through the dense structure of cartilage tissue surface;

Method used

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  • Application of nitrogen-doped carbon quantum dot delivery system in cartilage tissue
  • Application of nitrogen-doped carbon quantum dot delivery system in cartilage tissue
  • Application of nitrogen-doped carbon quantum dot delivery system in cartilage tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Nitrogen-doped carbon quantum dot delivery system, o-phenylenediamine reacts at high temperature and high pressure to generate nitrogen-doped carbon quantum dots;

[0038] The specific preparation method is: (1) Dissolve 300mg of o-phenylenediamine in 10ml of deionized water, and stir magnetically at room temperature to form a clear and transparent solution; (2) Add the solution into a polytetrafluoroethylene autoclave, and heat the muffle furnace to 180 ℃, reacted for 12 hours, cooled naturally to room temperature, and obtained carbon quantum dots; (3) filtered the obtained carbon quantum dots with a 0.22 μm filter membrane to remove large particles of impurities, and then dialyzed for 24 hours with a dialysis bag with a molecular weight cut-off of 3500 Da; (4) The carbon quantum dots after dialysis were freeze-dried and dissolved in deionized water to form a nitrogen-doped carbon quantum dot delivery system (concentration: 10 μg / ml).

Embodiment 2

[0039] Example 2: Detection of physical and chemical properties of nitrogen-doped carbon quantum dots

[0040] (1) Transmission electron microscope (TEM)

[0041] The prepared carbon quantum dots were diluted, dropped on the ultra-thin copper grid and dried, and the size, shape and distribution of the carbon quantum dots were observed by TEM.

[0042] The test shows that: figure 1 As shown, the prepared m-CQDs are spherical nanoparticles with good dispersion and relatively uniform size. The particle size distribution diagram is obtained by measuring the particle size of 100 carbon quantum dots, and the average size of carbon quantum dots is calculated as 4nm, and presents a normal distribution.

[0043] (2) Fluorescence spectrum

[0044] The optical characteristics of carbon quantum dots are analyzed by a fluorescence spectrophotometer, including excitation spectra in water, emission spectra under different excitation light and emission spectra of carbon dots at different p...

Embodiment 3

[0048] Example 3: m-CQDs were transfected into chondrocytes in vitro, and their transfection efficiency, biocompatibility, targeting, fluorescent signal tracking and metabolism were detected

[0049] 1. Detection of transfection rate:

[0050] A. Human chondrocyte extraction and culture process

[0051] 1) Knee joint tissue after joint replacement, choose a relatively normal part, and chop it into a mud in DMEM and mixed antibiotics (1%).

[0052] 2) The crushed cartilage tissue was washed once with DMEM, centrifuged at 1200 rpm for 5 minutes, and the washing solution was discarded.

[0053] 3) Add pronase (2mg / ml, HBSS configuration, add 1% mixed antibiotics) to the cartilage tissue at 37°C, 250 times / min, and incubate for 30 minutes.

[0054] 4) Discard the digestion solution, add DMEM to wash once, centrifuge at 1200 rpm for 5 minutes, and discard the washing solution.

[0055] 5) Add type II collagenase (0.1% - 0.3%, 2% FBS medium, add 1% mixed antibiotics) to cartilage...

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Abstract

The invention belongs to the technical field of biological medicine, and provides application of a nitrogen-doped carbon quantum dot delivery system in cartilage tissue in order to solve the problem that in current cartilage disease research, a compact structure on the surface layer of the cartilage tissue cannot be effectively penetrated to enter deep cells of the cartilage tissue. The application comprises the following steps of dissolving o-phenylenediamine in deionized water, and magnetically stirring at room temperature to form a clear and transparent solution; heating the solution to 180 DEG C in a high-pressure reaction kettle, reacting for 12 hours, and naturally cooling to room temperature to obtain carbon quantum dots; filtering by using a 0.22 m filter membrane of the carbon quantum dots, and dialyzing for 24 hours by using a dialysis bag with the molecular weight cut-off of 3500 Da; and freeze-drying, and dissolving in deionized water to obtain the nitrogen-doped carbon quantum dot delivery system. The nitrogen-doped carbon quantum dot delivery system is good in biocompatibility, high in targeting property, high in transfection efficiency, relatively small in size and capable of smoothly passing through a compact structure on the surface of the cartilage tissue, and can become an excellent cartilage tissue disease diagnosis method; and the system can smoothly enter the cartilage cells through the superficial compact structure of the cartilage tissue, and great clinical treatment significance is achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of a nitrogen-doped carbon quantum dot delivery system in cartilage tissue. Background technique [0002] Articular cartilage disease is a common and frequently-occurring disease that seriously endangers human health. The main pathological manifestations are joint swelling, pain, and dysfunction. It is one of the main causes of disability in middle-aged and elderly people [1]. The World Health Organization lists articular cartilage disease as the most widespread problem to the human body after the "three major killers" of cardiovascular and cerebrovascular diseases, cancer, and diabetes [2]. Therefore, it is extremely necessary to pay attention to and strengthen the research on articular cartilage diseases, which has a positive impact and important practical significance for improving people's quality of life, controlling medical expenses, and acce...

Claims

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Application Information

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IPC IPC(8): A61K47/04A61P19/04A61K49/00
CPCA61K47/02A61P19/04A61K49/0019
Inventor 郭丽段倩倩李鹏翠张博叶黄凌岸薛娟娟吴改革桑胜波卫小春
Owner THE SECOND HOSPITAL OF SHANXI MEDICAL UNIV
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