Nano-gene vector complex applied to directional transfection of inner ear hair cells and preparation method of nano-gene vector complex

A technology of inner ear hair cells and gene carriers, applied in the field of biomedicine, can solve the problems of ineffective combination and complete encapsulation of DNA, gene transfection efficiency related to molecular weight, high charge density, cytotoxicity, etc., so as to avoid low transfection efficiency and prepare The effect of easy process control, good biocompatibility and degradability

Inactive Publication Date: 2017-10-10
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, synthetic cationic polymers such as PEI have certain toxicity, and their gene transfection efficiency and toxicity are related to their molecular weight
High-molecular-weight PEI (such as 25kDa PEI) has more positive charges, so it has a higher ability to bind DNA and has a higher transfection efficiency, but because PEI is not degradable in vivo, the excessive charge Density also results in greater cytotoxicity, limiting in vivo applications
Low-molecular-weight PEI (such as <2kDa PEI) is less toxic, but it also has less positive charges, cannot effectively bind and completely encapsulate DNA, has poor carrier stability, and low transfection efficiency

Method used

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  • Nano-gene vector complex applied to directional transfection of inner ear hair cells and preparation method of nano-gene vector complex
  • Nano-gene vector complex applied to directional transfection of inner ear hair cells and preparation method of nano-gene vector complex
  • Nano-gene vector complex applied to directional transfection of inner ear hair cells and preparation method of nano-gene vector complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The method for preparing the nano-gene carrier complex for directional transfection of inner ear hair cells comprises the following steps:

[0036]1) Preparation of nano-gene complexes: Using the principle of electrostatic binding force, the Atoh1-EGFP plasmid and PEI nanoparticles were miscible in fully sterilized enzyme-free deionized water according to N / P=10:1. It is easy to be destroyed under the environment, so use a pipette to gently blow repeatedly 20-30 times, and react at room temperature for 20-30 minutes to obtain a nanocomposite with a particle size between 90-200nm and an obvious positive charge on the surface particles.

[0037] 2) Modification of the nano-gene complex: the nano-gene complex is modified by using biodegradable and highly compatible hyaluronic acid (Hyaluronic acid, HA). Add the PEI / Atoh1-EGFP complex into the HA solution according to COOH / N / P=4:10:1, gently blow and beat 20-30 times with a pipette, and react at room temperature for 20-30 ...

Embodiment 2

[0044] The steps of the method for preparing the nano-gene carrier complex for directional transfection of inner ear hair cells to treat inner ear hair cell damage are the same as in Example 1.

[0045] Example 2 The prepared nano-gene complex was transfected into the inner ear of the noise-induced deafness guinea pig model animal through the round window membrane permeation pathway, showing the repair effect on damaged hair cells after transfection.

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Abstract

The invention relates to the technical field of biological medicine, and in particular to a nano-gene vector complex applied to directional transfection of inner ear hair cells and a preparation method of the nano-gene vector complex. The nano-gene vector complex applied to directional transfection of inner ear hair cells consists of a hyaluronic acid modified PEI vector and a vector-loaded Atoh1 gene. The PEI vector is bPEI nanoparticles of 25kDa. The Atoh1 gene is derived from mice. According to the synthesized nano-gene vector complex provided by the invention, HA enveloped on the complex is excellent in biocompatibility and biodegradablity after the complex enters cells, and the generation of additional toxicity is prevented after in-vivo degradation; the nano-gene complex is protected from being bound by protein in body fluid in a transfection process, so that a low transfection efficiency is caused; and the complex prepared by the invention can effectively implement directional transfection of inner ear hair cells.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a nano-gene carrier complex for directional transfection of inner ear hair cells and a preparation method thereof. Background technique [0002] Sensorineural deafness is mainly caused by the degeneration of the auditory receptor - the cochlea. Studies on the anatomy of the human temporal bone have found that sensorineural deafness can be caused by the degeneration of various cell types in the cochlea. Among them, the hair cell lesion responsible for converting sound signals into electrical signals is the core issue in the current research on sensorineural deafness. one. Previously, researchers believed that in mammals, cochlear hair cells could not regenerate after injury and degeneration. [0003] In recent years, studies have found that introducing a plasmid containing the key gene for hair cell differentiation --- Atoh1 gene into the organ of Corti cultured in vitro can...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N15/85A61K48/00A61P27/16
CPCA61K48/0041C12N15/85C12N2800/107
Inventor吴南杨仕明王方园邓雄威吴雁陈志婷
OwnerGENERAL HOSPITAL OF PLA