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Manufacture method of decellularized trachea stent

A tracheal stent and decellularization technology, which is applied in the field of preparation of decellularized tracheal stents, can solve the problems that incomplete removal of tracheal cartilage cells, cells and antigen components affects the quality of life, and reduces the mechanical properties of the trachea. Reduced digestion time, good growth effect

Active Publication Date: 2021-03-19
GUANGDONG GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After 17 cycles of detergent-enzyme digestion (sodium deoxycholate and DNase I for about 35 days), the bioengineered tracheal matrix showed mechanical properties similar to those of the organism, allografts and xenografts There was no immune response in the next 30 days, showing that it has great potential for the replacement of tissue engineered trachea in the future, but this method cannot completely remove tracheal chondrocytes, and the residual nuclei still exist in the cartilage tissue. long
[0004] In conclusion, although the current use of decellularized trachea as a scaffold is feasible and effective, the decellularized tracheal matrix still faces long decellularization time and reduced tracheal mechanical properties after decellularized trachea with different chemical components. , Decellularization is not complete, residual cells and antigenic components may still affect the quality of life due to host immune rejection in the long-term prognosis

Method used

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  • Manufacture method of decellularized trachea stent
  • Manufacture method of decellularized trachea stent
  • Manufacture method of decellularized trachea stent

Examples

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Embodiment 1

[0061] A reagent for decellularization of animal trachea, including digestive juice A, digestive juice B, digestive juice C and digestive juice D,

[0062] The digestive juice A is EDTA digestive juice containing trypsin, and the mass percentage of trypsin in the digestive juice A is 0.25%;

[0063] The digestive solution B is a Tris-HCl buffer solution containing NaCl, the pH of the digestive solution B is 8.0, the concentration of Tris in the digestive solution B is 0.05M, and the NaCl concentration is 87.7g / L;

[0064] The digestive solution C is a Tris-HCl buffer containing TritonX-100, the pH of the digestive solution C is 7.4, the concentration of Tris in the digestive solution C is 0.05M, and the concentration of TritonX-100 is 10mL / L;

[0065] The digestive solution D contains DNAseI, RNAseI, MgCl 2 , CaCl 2 Tris-HCl buffer solution, the pH of the digestive solution D=7.6, the concentration of Tris in the digestive solution D is 0.01M, the concentration of DNAseI is ...

Embodiment 2

[0072] A method for preparing a decellularized tracheal stent, comprising the following steps:

[0073] (1) Acquisition and treatment of trachea

[0074] Put New Zealand white rabbits to death by injecting ketamine (25mg / kg) through the ear vein, put them on the ultra-clean aseptic workbench, put them on the center of the workbench in a supine position, disinfect the head, neck and chest with 75% ethanol, and use aseptic surgical instruments in sequence Separate the skin, fascia, and muscles to fully expose the trachea, cut the trachea about 2-4 cm along the cricoid cartilage at about 0.5 cm, put it in a culture dish containing PBS buffer, remove foreign objects on the surface of the trachea, and pay attention to maintaining the integrity of the trachea structure , soaked in double antibodies (penicillin and streptomycin) for 5 minutes, sterilized with 75% alcohol for 1 hour, put into cold D-Hanks buffer containing double antibodies, and washed 3 times on a shaker at 4°C (160 ...

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Abstract

The invention discloses a manufacture method of a decellularized trachea stent. Through a 0.25% of pancreatin-EDTA combined with detergent-ribozyme digestion method, the decellularized trachea stent is manufactured, a result indicates that a trachea matrix microstructure is completely kept, cells and residues in the trachea stent can be removed, in addition, an elimination effect is good, an immunological rejection reaction is small, and mechanical properties are good. Through the manufacture method disclosed by the invention, a decellularized trachea matrix can be obtained in about 7-10 days,digestion time is shortened, in addition, cell anchorage-dependent growth is good after in vitro implantation cell culture is carried out for three days, mechanical detection and biocompatibility detection do not have an obvious difference, and the decellularized trachea stent can serve as a choice of a tissue engineering trachea stent.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a method for preparing a decellularized tracheal stent. Background technique [0002] Tissue engineering is a new field that could potentially provide therapeutic alternatives for addressing defective organs without the risk of immune rejection and immunosuppression. At present, decellularized matrices are mainly used as potential scaffolds to obtain functional tissue engineered organs. While maintaining the integrity of the extracellular matrix and providing sufficient mechanical properties to maintain airway ventilation, tracheal grafts require a series of treatments to remove antigenicity and avoid immune rejection. [0003] Common methods for decellularized scaffold preparation include lyophilization and detergent methods. Low-pressure freeze-drying treatment of the trachea can destroy the proteoglycan and collagen components that make up the cartilage that maint...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3687A61L27/3691A61L27/3604A61L27/3679A61L27/50A61L2430/40A61L2430/22
Inventor 何少茹陈亮
Owner GUANGDONG GENERAL HOSPITAL