Preparation method and application of antibacterial damage repair microneedle for slowly releasing MSC (mesenchymal stem cell) sourced exosome

A technology of damage repair and exosomes, which is applied in the direction of microneedles, antibacterial drugs, and devices introduced into the body, can solve the problems of unstable treatment effect and large infusion dose, and improve the oxidative and inflammatory environment of the wound, blood medicine The effect of stable concentration and reduced frequency of administration

Inactive Publication Date: 2021-02-05
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a preparation method and application of an antibacterial damage repair microneedle that slowly releases MSC-derived exosomes in view of the shortcomings of traditional MSCs-exo in situ treatment, such as large infusion dose and unstable therapeutic effect.

Method used

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  • Preparation method and application of antibacterial damage repair microneedle for slowly releasing MSC (mesenchymal stem cell) sourced exosome
  • Preparation method and application of antibacterial damage repair microneedle for slowly releasing MSC (mesenchymal stem cell) sourced exosome
  • Preparation method and application of antibacterial damage repair microneedle for slowly releasing MSC (mesenchymal stem cell) sourced exosome

Examples

Experimental program
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Effect test

Embodiment 1

[0039] The preparation of antimicrobial repairing microneedles with slow-release MSC-exo, the steps are as follows:

[0040] (1) Preparation of gelatin methacrylate (GelMA) aqueous solution:

[0041] Weigh GelMA, add to double distilled water to disperse, prepare 30% GelMA solution, fully dissolve;

[0042] (2) Isolation of exosomes derived from mesenchymal stem cells:

[0043] Select MSCs in the logarithmic growth phase for the isolation and extraction of exosomes, and use MSC complete medium for culture. When the cells grow to 80% confluence, replace the MSC complete medium with serum-free exosome medium , continue culturing for 48 hours; collect the cell culture supernatant, and extract exosomes by ultracentrifugation. The ultracentrifugation conditions are: 50,000 rpm, 4°C, 60 min;

[0044] (3) Preparation of microneedle units loaded with MSC exosomes:

[0045] The exosomes derived from the 4th generation MSC were uniformly mixed at 50 μg / ml in the GelMA solution contai...

Embodiment 2

[0049] The preparation of antimicrobial repairing microneedles with slow-release MSC-exo, the steps are as follows:

[0050] (1) Preparation of methacrylate hyaluronic acid aqueous solution:

[0051] Weigh a certain amount of methacrylate hyaluronic acid, add it to double-distilled water to disperse, prepare a 40% aqueous solution, and fully dissolve it;

[0052] (2) Isolation of exosomes derived from mesenchymal stem cells:

[0053] Select MSCs in the logarithmic growth phase for the isolation and extraction of exosomes, and use MSC complete medium for culture. When the cells grow to 80% confluence, replace the MSC complete medium with serum-free exosome medium , continue culturing for 48 hours; collect the cell culture supernatant, and extract exosomes by ultracentrifugation. The ultracentrifugation conditions are: 60,000 rpm, 4°C, 120 min;

[0054] (3) Preparation of microneedle units loaded with MSC exosomes:

[0055] The exosomes derived from the third-generation MSCs ...

Embodiment 3

[0059] The preparation of antimicrobial repairing microneedles with slow-release MSC-exo, the steps are as follows:

[0060] (1) Preparation of gelatin methacrylate (GelMA) aqueous solution:

[0061] Weigh GelMA, add to double distilled water to disperse, prepare 35% GelMA solution, fully dissolve;

[0062] (2) Isolation of exosomes derived from mesenchymal stem cells:

[0063] Select MSCs in the logarithmic growth phase for the isolation and extraction of exosomes, and use MSC complete medium for culture. When the cells grow to 80% confluence, replace the MSC complete medium with serum-free exosome medium , continue culturing for 48 hours; collect the cell culture supernatant, and extract exosomes by ultracentrifugation. The ultracentrifugation conditions are: 65,000 rpm, 4°C, 60 min;

[0064] (3) Preparation of microneedle units loaded with MSC exosomes:

[0065] The exosomes derived from the 4th generation MSC were evenly mixed in the GelMA solution containing 1% (v / v) p...

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Abstract

The invention provides an antibacterial damage repair microneedle for slowly releasing MSC (mesenchymal stem cell) sourced exosome. The microneedle comprises a substrate and a microneedle unit fixed to the substrate, wherein the substrate is a soluble high-molecular polymer doped with silver nitrate, and the microneedle unit is a biodegradable material loaded with the MSC sourced exosome. According to the invention, a PDMS mold prepared with an etching technology is adopted to construct the damage repair microneedle with the microneedle unit loaded with MSC-exo and a substrate containing antibacterial metal nanoparticles, and after the damage repair microneedle acts on the skin, the MSC-exo can be slowly released to play anti-inflammatory and immunomodulatory roles, accelerate angiogenesisand improve the oxidative inflammation environment of the wound surface; meanwhile, the antibacterial metal nanoparticles in the substrate have the effect of preventing and treating bacterial infection, chronic wound healing of the skin is accelerated, the minimum treatment dosage of the MSC-exo and the antibacterial metal nanoparticles can be achieved in situ, and painless, safe and autonomous administration is achieved; and the preparation process is simple, large instruments are not needed, the medicine carrying capacity is high, safety and practicability are high, and popularization is easy.

Description

technical field [0001] The invention belongs to the technical field of drug-loaded release of microneedles, and in particular relates to a preparation method and application of an antibacterial damage repairing microneedle for slow release of MSC-derived exosomes. Background technique [0002] Diabetes mellitus is a metabolic disease with a high incidence rate. Abnormal accumulation of blood sugar leads to pathological processes such as neuropathy and arterial damage, which affects the health and quality of life of patients for a long time. Hyperglycemia-induced microvascular dysfunction is one of the main causes of diabetic complications, leading to wound ischemia and delayed healing. Wound healing is generally divided into three classic stages of inflammation, proliferation, and matrix remodeling, which require blood vessels to provide nutrients and oxygen support to the cells involved in the healing process. Therefore, enhancing angiogenesis and remodeling in diabetic wo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K33/38A61K9/00A61K47/36A61K47/42A61P17/02A61P31/04A61P3/10A61M37/00
CPCA61K35/28A61K33/38A61K9/0021A61K47/36A61K47/42A61P17/02A61P31/04A61P3/10A61M37/0015A61M2037/0046A61M2037/0053
Inventor 赵远锦甘璟璟王月桐商珞然
Owner NANJING DRUM TOWER HOSPITAL
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