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A preparation method of microenvironment-responsive immune regulation promoting nerve regeneration micro-nanofibers

A micro-nano fiber, immune regulation technology, applied in fiber treatment, tissue regeneration, pharmaceutical formulations, etc., can solve problems such as low concentration and excessive drug loss, and achieve the goal of promoting angiogenesis, reducing inflammation, and reducing scar tissue formation. Effect

Active Publication Date: 2021-10-12
海南德拉米克投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Oriented electrospun fibers with biomimetic structures have been used in the research of nerve tissue repair for many times, but electrospinning still has problems such as sudden release of loaded drugs, rapid drug loss, and low concentration.

Method used

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  • A preparation method of microenvironment-responsive immune regulation promoting nerve regeneration micro-nanofibers
  • A preparation method of microenvironment-responsive immune regulation promoting nerve regeneration micro-nanofibers
  • A preparation method of microenvironment-responsive immune regulation promoting nerve regeneration micro-nanofibers

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation method of microenvironment-responsive immune regulation and promoting nerve regeneration comprises the following steps:

[0040] (1) Preparation of formylated cationic liposomes

[0041] The reverse evaporation method was used to prepare the aldylated cationic liposomes. First, 160 mg of soybean lecithin, 40 mg of cholesterol, and 4 mg of aldophospholipids were dissolved in 5 mL of chloroform, and 5 mg of octadecylamine was weighed and dissolved in 1 mL of chloroform. After mixing, carry out ultrasonication to obtain a uniform emulsion, then remove the organic solvent in a rotary evaporator to obtain a colloidal product, and finally add deionized water for hydration treatment to prepare a liposome emulsion. Filtrate to obtain formylated cationic liposomes.

[0042] (2) Preparation of liposomes loaded with eGFP-IL-4 plasmid

[0043] Prepare five 1mL centrifuge tubes, numbered A-E, add 100 μL Opti-MEM medium to each tube, and then add eGFP-IL-4 plasmid (...

Embodiment 2

[0055] 1. Physicochemical properties characterization and screening of aldylated cationic liposomes

[0056] 1. Transmission Electron Microscopy (TEM)

[0057] Take 2 μL of formylated cationic liposomes and drop them on the copper grid, and dry them at room temperature for 4 h. Apply a transmission electron microscope (TEM, Hitachi HT7700) to observe the surface morphology of the formylated cationic liposomes at a voltage of 120 kV.

[0058] 2. Detection of particle size and surface charge of aldehylated cationic liposomes

[0059] Take 2mL cationic liposome and plasmid composite solution of each ratio respectively, and use a dynamic light scattering particle size analyzer (DLS) to detect the size, polydispersity index, and kinetic potential.

[0060] 3. Encapsulation rate detection

[0061] Add 1mL cationic liposome and plasmid composite solution samples into ultrafiltration centrifuge tubes respectively, set the centrifuge speed to 5000rpm, and centrifuge for 5min. Then a...

Embodiment 3

[0096] Example 3 Growth and differentiation of cells on the surface of micro-nano fibers

[0097] 1. Preparation of cells for research

[0098] Before cell research, fibrous scaffolds collected on coverslips with a diameter of 15 mm and a thickness of 100 μm were gently placed on the bottom of the Transwell plate and sterilized by irradiation, and then grafted with aldehylated cationic lipids loaded with pDNA as described above Plastids were placed in a 37°C cell culture incubator. According to the density of 1×10 4 SD rat bone marrow mesenchymal stem cells (BMSCs) per hole were planted on the fiber scaffold, and the planting density of the upper chamber was 5×10 3 SD rat bone marrow macrophages (BMM) per well, put the Transwell plate at 37°C, relative humidity 95%, CO 2 Co-cultivation was performed in a partial pressure 5% incubator. The medium was changed every 2-3 days. The non-fibrous scaffold group (Control), traditional fibrous scaffold (aP), microsol fibrous scaffo...

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Abstract

The invention discloses a preparation method of microenvironment-responsive immunoregulation and nerve regeneration-promoting micro-nanofibers, comprising the following steps: (1) preparation of aldehyde-formylated cationic liposomes; (2) preparation of eGFP-IL-4 plasmids Preparation of liposomes; (3) Preparation of directional electrospun fiber membranes; (4) Preparation of micro-nanofibers for microenvironment-responsive immune regulation and promoting nerve regeneration. The micro-nanofiber can reduce inflammatory response, down-regulate the secretion of glial fibrillary acidic protein, reduce the formation of scar tissue, promote angiogenesis, and continuously release NGF to promote the neural differentiation ability and functional recovery of endogenous stem cells. Therefore, the micro-nanofiber is an innovative responsive sequential immune regulation and neural regeneration-promoting functional organism that preferentially regulates the immune regulation of the local microenvironment of spinal cord injury, and then provides a neural differentiation platform for endogenous stem cells for therapeutic purposes. Scaffold, which provides a new strategy for tissue engineering to treat spinal cord injury.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and in particular relates to a preparation method of a micro-environment-responsive immune regulation and nerve regeneration-promoting micro-nano fiber. Background technique [0002] Spinal cord injury leads to permanent changes or even loss of sensory and motor functions below the level of injury, which is still an insurmountable problem in the progress of human medicine. After primary spinal cord injury, local tissue necrosis, inflammatory infiltration, and free radical oxidative stress caused H + Elevated concentration forms an acidic environment to further damage the surrounding tissues, and at the same time, the invasion of inflammatory factors causes secondary spinal cord injury and gradually aggravates the degree of injury. Secondary spinal cord injury involves far more lesions than primary spinal cord injury, in which microglia activated in the central nervous system and per...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/54A61L27/50A61L27/22D04H1/728D04H3/02D01D5/00
CPCA61L27/227A61L27/50A61L27/54A61L2300/252A61L2300/414A61L2300/426A61L2300/602A61L2400/12A61L2430/32D01D5/003D04H1/728D04H3/02C08L89/00
Inventor 陈亮郗焜施勤顾勇崔文国
Owner 海南德拉米克投资有限公司
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