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Non-virus nano nucleic acid transferring composite for curing gristle defection by injecting in joint cavity and preparing method thereof

A compound and joint cavity technology, applied in the field of non-viral nano-nucleic acid transport complex and its preparation, can solve the problems of high solution viscosity, large molecular weight, and limited application, so as to reduce viscosity, increase water solubility, and enhance biocompatibility sexual effect

Inactive Publication Date: 2011-03-23
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, the molecular weight of undegraded high molecular weight chitosan (High Molecular Weight Chitosan, HMWC) is too large, it is difficult to dissolve in water, and needs to be dissolved in acid, and the viscosity of the formed solution is relatively high. Strong, and the damage to cells is also relatively large, which limits its direct application as a gene transfer carrier

Method used

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  • Non-virus nano nucleic acid transferring composite for curing gristle defection by injecting in joint cavity and preparing method thereof
  • Non-virus nano nucleic acid transferring composite for curing gristle defection by injecting in joint cavity and preparing method thereof
  • Non-virus nano nucleic acid transferring composite for curing gristle defection by injecting in joint cavity and preparing method thereof

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Experimental program
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Effect test

Embodiment 1

[0053] The preparation of embodiment 1 low molecular weight chitosan

[0054]Get 5g commercially available high molecular weight chitosan (St.Louis, MO, USA, molecular weight is about 150KDa, 85% deacetylation) in the 250ml round bottom flask, add the 4M HCl of 125ml, oil bath 100 ℃ 15 hours, simultaneously Stir. After cooling, filter with double-layer lens-cleaning paper to obtain 85ml of solution, add an equal volume of absolute ethanol to the solution, mix well, and freeze overnight at 4°C. Then centrifuge at 10000rpm for 10 minutes and discard the supernatant. Add 50ml of 50% ethanol to the pellet to wash, resuspend, and centrifuge at 10000rpm for 10 minutes. Repeat the above washing, suspension and centrifugation process three times. The precipitate was dried, dissolved in 15 ml of double distilled water, and finally freeze-dried to obtain a white powdery solid, which was low molecular weight chitosan. The composition of the product was determined by elemental analysi...

Embodiment 2

[0055] The preparation of embodiment 2 low molecular weight chitosan

[0056] Get 5g commercially available high molecular weight chitosan (St.Louis, MO, USA, molecular weight is about 150KDa, 85% deacetylation) in the 250ml round bottom flask, add the 4M HCl of 140ml, oil bath 100 ℃ 25 hours, simultaneously Stir. After cooling, filter with double-layer lens-cleaning paper to obtain 90 ml of solution, add an equal volume of absolute ethanol to the solution, mix well, and freeze overnight at 4°C. Then centrifuge at 10000rpm for 10 minutes and discard the supernatant. Add 50ml of 50% ethanol to the pellet to wash, resuspend, and centrifuge at 10000rpm for 10 minutes. Repeat the above washing, suspension and centrifugation process three times. The precipitate was dried, dissolved in 15 ml of double distilled water, and finally freeze-dried to obtain a white powdery solid, which was low molecular weight chitosan. The composition of the product was determined by elemental analy...

Embodiment 3

[0057] The preparation of embodiment 3 low molecular weight chitosan

[0058] Get 5g commercially available high molecular weight chitosan (St.Louis, MO, USA, molecular weight is about 150KDa, 85% deacetylation) in the 250ml round bottom flask, add the 4M HCl of 110ml, oil bath 100 ℃ 10 hours, simultaneously Stir. After cooling, filter with double-layer lens-cleaning paper to obtain 90 ml of solution, add an equal volume of absolute ethanol to the solution, mix well, and freeze overnight at 4°C. Then centrifuge at 10000rpm for 10 minutes and discard the supernatant. Add 50ml of 50% ethanol to the pellet to wash, resuspend, and centrifuge at 10000rpm for 10 minutes. Repeat the above washing, suspension and centrifugation process three times. The precipitate was dried, dissolved in 15 ml of double distilled water, and finally freeze-dried to obtain a white powdery solid, which was low molecular weight chitosan. The composition of the product was determined by elemental analy...

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Abstract

The invention discloses a non-viral nanometer nucleic acid transportation compound used for treating cartilage defect through intra-articular injection which comprises low-molecular chitosan with molecular weight of 5KD-100KDand percentage composition of amino group of 30-70%, and nucleic acid, wherein N / P of the compound is 0.2:1-10:1.The invention degrades high molecular chitosan to low molecular chitosan, increases water solubility of material, reduces viscosity of solution, reduces influence of sour environment of solvent to cell, improves compound size with nanometer, is in favor of performing genetic transmission to cartilago articularis cell, and protects target nucleic acid from neutralizing with synovial fluid and being degrade by nuclease in lysosome. The invention has simple preparation and abroad application scope; can solve engineering roadblock of cartilage repair with no requirement of operation, complex device and bracket material.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to a non-viral nano-nucleic acid transport complex used for intra-articular injection to treat cartilage defects and a preparation method thereof. Background technique [0002] Articular cartilage defect is one of the common and difficult clinical diseases, which can be caused by trauma, inflammation, tumor and autoimmune diseases and other reasons. After the defect occurs, it is difficult to repair. This is because the articular cartilage is hyaline cartilage, which has a special structure different from other cartilages. It is mainly composed of chondrocytes embedded in the lacuna of the cartilage matrix, and is a single connective tissue. The cartilage matrix is ​​mainly synthesized and maintained by chondrocytes, and its main components are various proteoglycans and collagen fibers mainly composed of type II collagen. Articular cartilage tissue has neithe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/36A61K48/00A61P19/08
Inventor 赵建宁王瑞郭亭张峻峰
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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