Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Novel dermal matrix decellularization method

A dermal matrix and decellularization technology, applied in medical science, prostheses, etc., can solve the problems of reduced mechanical properties of materials, structural damage of dermal matrix, poor biocompatibility, etc., to improve stability, induce cell growth, increase Effects of hydrophilicity and cell affinity

Inactive Publication Date: 2020-07-24
百澳瑞派(天津)生物科技有限公司
View PDF12 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The commonly used decellularization methods include physical methods (such as stirring, shaking, ultrasound, hypotonicity and hypertonicity), chemical methods (such as acids, alkalis, surfactants), enzymatic methods (such as trypsin, DNase, etc.), organic solvents, etc. (such as ethanol, acetone, etc.), these four methods will cause different degrees of damage to the structure of the dermal matrix, degrade the active factors in the matrix, and lead to a decrease in the mechanical properties of the material, poor thermal stability, and poor biocompatibility.

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
  • Novel dermal matrix decellularization method
  • Novel dermal matrix decellularization method
  • Novel dermal matrix decellularization method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (1) Take fresh calf leather (purchased from the slaughterhouse), use a leather machine to remove the epidermis and subcutaneous tissue of the cowhide, and obtain a 0.5mm dermal reticular layer, soak it in 0.1% bromogeramine solution immediately, sterilize and degrease After 30 minutes, wash with PBS.

[0036] (2) Use purified water to prepare PBS buffer solution containing structural stabilizer 10% silk fibroin, 2.5% arginine and 1% sucrose, shake and mix at 120 rpm / min for 10 min at room temperature, and filter to sterilize with a 0.45 μm water filter. Place the bovine leather prepared above in the mixed solution, soak at 4°C for 30 minutes under low pressure, the pressure is 300Pa, the soaking ratio is solution volume: wet weight of dermal matrix = 1:1 (w / v).

[0037] (3) Use purified water to prepare a decellularized mixture containing 10% silk fibroin, 2.5% arginine, 1% sucrose, 0.25% trypsin, 0.15% ficin and PBS, shake and mix at 120rpm / min for 15min at room temper...

Embodiment 2

[0045] (1) Take fresh pigskin (purchased from a slaughterhouse), remove the epidermis and subcutaneous tissue of the pigskin with a skin remover, and obtain a 0.7mm dermal reticular layer, soak it in 0.1% bromogeramine solution immediately, and sterilize and degrease for 30 minutes , washed with PBS.

[0046] (2) Use purified water to prepare PBS buffer solution containing structural stabilizers 2.5% mannitol, 1% glycine, and 1% sucrose, shake and mix at room temperature for 5 minutes at a speed of 100 rpm / min, and filter to sterilize with a 0.45 μm water filter. Put the above-prepared porcine dermis in the mixed solution, soak for 1 hour at 4°C under low pressure, the pressure is 500Pa, the soaking ratio is solution volume: wet weight of dermal matrix = 1:1 (w / v).

[0047] (3) Use purified water to prepare a decellularized mixture containing 3% mannitol, 1% glycine, 1% sucrose, 0.5% SDS, 2% TrionX-100 and PBS, shake and mix at 180rpm / min for 30min at room temperature, 0.45μm ...

Embodiment 3

[0052] (1) Take fresh pigskin (purchased from a slaughterhouse), remove the epidermis and subcutaneous tissue of the pigskin with a skin remover, and obtain a 0.7mm dermal reticular layer, soak it in 0.1% bromogeramine solution immediately, and sterilize and degrease for 30 minutes , washed with PBS.

[0053] (2) Use purified water to prepare PBS buffer solution containing structural stabilizers 5% silk fibroin and 3% arginine, shake at room temperature for 5 minutes, rotate at 120 rpm / min, and filter to sterilize with a 0.45 μm water filter membrane. Place the pig dermis prepared above in the mixed solution, soak at 2°C for 20 minutes under low pressure, the pressure is 200 Pa, and the soaking ratio is solution volume: wet weight of dermal matrix = 2:1 (w / v).

[0054] (3) Use purified water to prepare a decellularized mixture containing 5% silk fibroin, 3% arginine, 0.25% trypsin, 1% TrionX-100 and PBS, shake and mix at 180rpm / min for 20min at room temperature, 0.45μm Steril...

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

No PUM Login to View More

Abstract

The invention relates to a novel dermal matrix decellularization method. Dermis is treated by solutions containing a structural stabilizer in the whole course, and the novel dermal matrix decellularization method comprises soaking before decellularization, tissue decellularization, and cleaning after decellularization, and comprises the following steps: (1) placing a dermal matrix in a PBS buffersolution containing the structural stabilizer, and conducting soaking at 2-8 DEG C and low pressure of 200-500 Pa for 20 min to 2 h; (2) placing the dermal matrix in a decellularization solution containing the structural stabilizer, and conducting shaking treatment at 2-8 DEG C and 80-200 rpm for 4-6 h; and (3) placing the dermal matrix in a decellularization solution containing the structural stabilizer, and conducting ultrasonic high-pressure treatment at 2-4 DEG C for 30-60 min, wherein the ultrasonic power is 20-40 Hz, and the pressure is 200-500 MPa. The structural stabilizer comprises two or three of silk fibroin, mannitol, glycine, arginine and sucrose. A dermal matrix material produced through the decellularization method has softer appearance and strength, and the thermal stability, the mechanical performance and retention of bioactive components are all improved, so that the hydrophilicity and the cellular affinity of the material are improved, and cell ingrowth can be induced.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and relates to a decellularization technology of dermal matrix, in particular to a novel decellularization method of dermal matrix. Background technique [0002] The skin is composed of epidermis, dermis and subcutaneous tissue. The dermis is a layer of dense connective tissue composed of collagen fibers, fibroblasts, smooth muscle cells, endothelial cells, and blood vessels. Collagen fibers are mainly composed of type Ⅰ collagen, with little difference among species, while foreign cells, interstitial fluid, nucleic acid, and lipid are the factors that cause immunogenicity. When the dermal matrix is ​​implanted into the human body as a foreign tissue, the above factors such as cells and nucleic acids need to be removed to avoid immune rejection. The dermis matrix can come from allogeneic skin, pigskin, cowhide, sheepskin, etc. [0003] The commonly used decellularization methods include phys...

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 Applications(China)
IPC IPC(8): A61L27/36
CPCA61L27/362A61L27/3687A61L27/3691A61L27/3695
Inventor 蒋彩云李志宏郭雅楠柏玮
Owner 百澳瑞派(天津)生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com