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Preparation method of externally constructed tissue engineering trachea matrix

A technology of tissue engineering and trachea, which is applied in the field of biomedical engineering, can solve the problems of poor tissue stability, difference in effect, unfavorable growth and shaping of host cells, etc., to reduce the reaction between different species, avoid calcification and immune reaction, and prevent excessive The effect of early degradation

Inactive Publication Date: 2015-12-23
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

However, there are also differences in the effect. The existing treatment methods are difficult to remove the tracheal cells and retain the complete tracheal matrix scaffolds such as cartilage scaffolds, collagen fibers and elastic fibers.
Tracheal mucosal epithelial cells are the main factor causing immune rejection of tracheal transplantation, so the existing decellularization methods are difficult to meet the requirements
In particular, the anti-degradation ability of extracellular matrix components is low, the tissue stability is poor, and it is easy to be absorbed in a short period of time after implantation, which is not conducive to the growth and shaping of host cells.

Method used

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  • Preparation method of externally constructed tissue engineering trachea matrix
  • Preparation method of externally constructed tissue engineering trachea matrix
  • Preparation method of externally constructed tissue engineering trachea matrix

Examples

Experimental program
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Effect test

Embodiment 1

[0034] (1) Under sterile conditions, obtain fresh allogeneic or heterogeneous trachea (such as porcine trachea) tissues that meet the requirements, and store them in phosphate buffered saline at 0-4°C.

[0035] (2) Trim under aseptic operation, remove adjacent fat and other connective tissue, and lavage with phosphate buffer solution containing antibiotics to remove blood on the tracheal tube.

[0036] (3) decellularization, the steps include:

[0037] ① Soak in 0.1wt% brogeramine for 30 minutes; place the trachea in 0.5wt% Triton X-100 (TritonX-100), and incubate at 37°C for 48 hours;

[0038] ② Place in 0.25wt% trypsin and 0.02wt% EDTA (volume ratio 1:1) and incubate for 30 minutes.

[0039] ③Incubate at 37°C for 24 hours with 30 μL / ml DNase-1 and 0.3 mg / mL RNase-A.

[0040] (4) photooxidative cross-linking treatment, the steps include:

[0041] ① Soak the decellularized trachea in hypertonic PBS solution (pH7.6, Osm: 680mosm) for 4 hours;

[0042] ② Place in PBS solutio...

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Abstract

The invention discloses a preparation method of an externally constructed tissue engineering trachea matrix. The preparation method comprises the following steps: pretreatments including fat removing and blood cleaning are performed on fresh allogeneic trachea tissue or xenogenic trachea tissue; then trachea tissue impurities and tissue cells are removed through the combination of a chemical method and a biological enzyme method, and finally, a photooxidation cross-linking reaction is adopted for adjusting the crosslink level of a trachea matrix framework of which the tissue cells are removed; and through the method, the tissue engineering trachea matrix which is good in stability and beneficial to seed cell seeding, attachment and proliferation can be obtained.

Description

technical field [0001] The invention relates to a preparation method of decellularized tissue engineering trachea matrix, which belongs to the field of biomedical engineering. Background technique [0002] The trachea often causes airway obstruction due to factors such as tumors or inflammatory lesions, which seriously endangers the lives of patients. Surgical resection of the diseased trachea is the most effective clinical treatment. However, due to the limitation of the extensibility of the trachea itself, long-segment tracheectomy must be performed with the help of tracheal substitutes for airway reconstruction. At present, there are mainly three kinds of tracheal substitutes, namely, allogeneic tracheal transplantation, autologous tissue reconstruction, and artificial trachea. The trachea is a hollow and elastic vital organ that maintains ventilation, cleaning and defense functions of the human body, and has unique anatomical and physiological characteristics. Tracheal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
Inventor 刘文亮肖鹏
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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