Preparation method of double-gene time sequence sustained-release tissue engineering scaffold material

A tissue engineering scaffold, dual-gene technology, applied in gene therapy, pharmaceutical formulations, genetic material management system, etc.

Active Publication Date: 2017-12-01
温州医科大学附属口腔医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

How to effectively exert the timely and appropriate effect of cell signaling factors, how to simulate the programmed release of multiple factors in the normal physiological process, and exert normal physiological functions to achieve the ultimate ideal functional replacement have not been effectively resolved.

Method used

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  • Preparation method of double-gene time sequence sustained-release tissue engineering scaffold material

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Step 1: Through plasmid construction and amplification, 0.1g rhBMP-2 and 0.1g rhIGF-I are obtained.

[0032] Step 2. Preparation of rhBMP-2DNA / polyacetimide nanoparticles:

[0033] a. Mix 0.1grhBMP-2DNA and 30g polyethylene oxide uniformly to obtain a drug mixture,

[0034] b. Dissolve the drug mixture in 50mL N,N-dimethylformamide to obtain a drug oil solution;

[0035] c. Add the drug oil solution to 90g of a polyacetimide ethanol solution with a mass concentration of 60% and a weight average molecular weight of 1000 and continue to stir for 30 minutes;

[0036] d. Add 40g sucrose and lyophilize for later use.

[0037] Step 3. Preparation of rhIGF-Ⅰ DNA / polyacetimide nanoparticles;

[0038] a. Mix 0.1g rhIGF-I and 30g erythritol uniformly to obtain a drug mixture,

[0039] b. Dissolve the drug mixture in 50 mL of tetrahydrofuran to obtain a drug oil solution;

[0040] c. Add the drug oil solution to 90 g of a polyacetimide ethanol solution with a mass concentration of 60% and a wei...

Embodiment 2

[0049] Step 1: Through plasmid construction and amplification, 0.1g rhBMP-2 and 0.1g rhIGF-I are obtained.

[0050] Step 2. Preparation of rhBMP-2DNA / polyacetimide nanoparticles;

[0051] a. Mix 0.1grhBMP-2DNA and 30g polypyrrolidone uniformly to obtain a drug mixture,

[0052] b. Dissolve the drug mixture in 50 mL of thionyl chloride to obtain a drug oil solution;

[0053] c. Add the drug oil solution to 90 g of a polyacetimide solution with a mass concentration of 60% and a weight average molecular weight of 3500 and continue to stir for 40 minutes;

[0054] d. Add 40g of sucrose and freeze-dry it for later use;

[0055] Step 3. Preparation of rhIGF-Ⅰ DNA / polyacetimide nanoparticles;

[0056] a. Mix 0.1g rhIGF-I and 30g pharmaceutical excipient polyethylene oxide uniformly to obtain a drug mixture,

[0057] b. Dissolve the drug mixture in 50 mL of organic solvent ethyl acetate to obtain a drug oil solution;

[0058] c. Add the drug oil solution to 90g of a polyacetimide solution with a ma...

Embodiment 3

[0067] Step 1: Through plasmid construction and amplification, 0.1g rhBMP-2 and 0.1g rhIGF-I are obtained.

[0068] Step 2. Preparation of rhBMP-2DNA / polyacetimide nanoparticles;

[0069] a. Mix 0.1grhBMP-2DNA and 30g erythritol as a pharmaceutical excipient to obtain a drug mixture.

[0070] b. Dissolve the drug mixture in 50 mL of organic solvent thionyl chloride to obtain a drug oil solution;

[0071] c. Add the drug oil solution to 90 g of a polyacetimide solution with a mass concentration of 60% and a weight average molecular weight of 9000 and continue to stir for 50 minutes;

[0072] d. Add 40g of sucrose and freeze-dry it for later use;

[0073] Step 3. Preparation of rhIGF-Ⅰ DNA / polyacetimide nanoparticles;

[0074] a. Mix 0.1g rhIGF-I and 30g polyethylene oxide uniformly to obtain a drug mixture,

[0075] b. Dissolve the drug mixture in 50mL N,N-dimethylformamide to obtain a drug oil solution;

[0076] c. Add the drug oil solution to 90 g of a polyacetimide solution with a mass co...

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Abstract

The invention discloses a preparation method of a double-gene time sequence sustained-release tissue engineering scaffold material. The preparation method comprises the following steps: firstly, encapsulating plasmid containing IGF-I by virtue of a sustained-release material which is relatively high in molecular weight, so that sustained-release microspheres are prepared; and preparing the three-dimensional porous tissue engineering material from the microspheres, PLGA which is relatively low in molecular weight and plasmid containing BMP-2 by virtue of a supercritical CO2 and particulate leaching method. As the material is implanted into a body, a main body material, which is relatively low in molecular weight, around the BMP-2 plasmid which is not encapsulated in sustained-release microspheres is degraded firstly, and the BMP-2 plasmid is released firstly; while the IGF-I plasmid, which is encapsulated in the material, which is relatively high in molecular weight, is delayed in release, so that an effect that two target genes are released in a certain order. According to the material, cell factors can be subjected to time sequence expression in a mode of simulating a physiological process of tissue repair, so that the material is more conducive to tissue regeneration and systemic side effects are reduced; a gene product can achieve local continuous release; and a local therapeutic effect can be enhanced to the greatest extent.

Description

Technical field [0001] The invention relates to the field of medical materials, and more specifically to a method for preparing a tissue engineering scaffold material with a sequential slow release of double genes. Background technique [0002] Bone tissue repair is a complex process involving a series of chain processes such as the differentiation and proliferation of pluripotent stem cells, the recognition of extracellular matrix and signal molecules, the expression and targeting of related factors, and the development and maturation of new bone. Various osteogenic factors such as bone morphogenetic protein family, fibroblast growth factor, transforming growth factor β, platelet-derived growth factor, vascular endothelial growth factor, insulin-like growth factor, etc., all play an important role in this process In addition, there is a certain phase, site law and synergy / antagonism relationship between them. Applying various osteogenic factors or controlled-release carriers lo...

Claims

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Application Information

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IPC IPC(8): A61L27/22A61L27/18A61L27/54A61L27/50A61L27/56A61L27/58A61K48/00
CPCA61K48/005A61K48/0083A61L27/18A61L27/22A61L27/50A61L27/54A61L27/56A61L27/58A61L2300/252A61L2300/258A61L2300/412A61L2300/602C08L79/02C08L67/04
Inventor 刘劲松徐丽华邓振南姚李韬吴星海平林超
Owner 温州医科大学附属口腔医院
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