A kind of preparation method of double gene sequential slow release tissue engineering scaffold material

A tissue engineering scaffold and double gene technology, applied in gene therapy, pharmaceutical formula, genetic material management system, etc.

Active Publication Date: 2020-07-14
温州医科大学附属口腔医院
View PDF4 Cites 0 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of preparation method of double gene sequential slow release tissue engineering scaffold material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Step 1: Obtain 0.1 g of rhBMP-2 and 0.1 g of rhIGF-I through plasmid construction and amplification.

[0032] Step 2, preparation of rhBMP-2DNA / polyacetimide nanoparticles:

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

[0034] b, dissolving the drug mixture into 50mL N,N-dimethylformamide to obtain a drug oil solution;

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

[0036] d. Add 40 g of sucrose and freeze-dry it for later use.

[0037] Step 3, preparation of rhIGF-I DNA / polyacetylimide nanoparticles;

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

[0039] b, the drug mixture is dissolved in 50mL tetrahydrofuran to obtain a drug oil solution;

[0040] c. Add the drug oil solution to 90 g of polyacetimide ethanol solution with a mass concentr...

Embodiment 2

[0049] Step 1: Obtain 0.1 g of rhBMP-2 and 0.1 g of rhIGF-I through plasmid construction and amplification.

[0050] Step 2, preparation of rhBMP-2DNA / polyacetimide nanoparticles;

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

[0052] B, the drug mixture is dissolved in 50mL 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 continuously stir for 40 min;

[0054] d, add 40g of sucrose and freeze-dry for subsequent use;

[0055] Step 3, preparation of rhIGF-I DNA / polyacetylimide nanoparticles;

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

[0057] B, the drug mixture is dissolved in 50mL organic solvent ethyl acetate to obtain drug oil solution;

[0058] c. Add the drug oil solution to 90 g of a polyethylimide so...

Embodiment 3

[0067] Step 1: Obtain 0.1 g of rhBMP-2 and 0.1 g of rhIGF-I through plasmid construction and amplification.

[0068] Step 2, preparation of rhBMP-2DNA / polyacetimide nanoparticles;

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

[0070] B, the drug mixture is dissolved in 50mL organic solvent thionyl chloride to obtain 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 continuously stir for 50 min;

[0072] d, add 40g of sucrose and freeze-dry for subsequent use;

[0073] Step 3, preparation of rhIGF-I DNA / polyacetylimide nanoparticles;

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

[0075] b, dissolving the drug mixture into 50mL N,N-dimethylformamide to obtain a drug oil solution;

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to view more

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 relates to a preparation method of a double-gene time-sequential slow-release tissue engineering scaffold material. Background technique [0002] Bone tissue repair is a complex process, involving a series of chain processes such as differentiation and proliferation of pluripotent stem cells, recognition of extracellular matrix and signaling molecules, expression and targeting of related factors, and 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. and there is a certain phase, site rule, and synergistic / antagonistic relationship between them. Applying various osteogenic factors or controlled-release carriers loaded...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products