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Method for preparing rabbit acellular tracheal matrix

A technology of decellularization and rabbit trachea, applied in the field of bioengineering, can solve the problems of unsuitable emergency tracheal graft preparation, long time-consuming and high cost

Active Publication Date: 2019-07-02
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it requires 7 preparation cycles, which is costly and time-consuming, so it is not suitable for the preparation of emergency tracheal grafts

Method used

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  • Method for preparing rabbit acellular tracheal matrix
  • Method for preparing rabbit acellular tracheal matrix
  • Method for preparing rabbit acellular tracheal matrix

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1.1 Selection and grouping of experimental animals

[0026] Select 30 healthy New Zealand rabbits of 2.5-3.5kg, male or female, and randomly divide them into 3 groups. Group A is the original control group, and groups B and C correspond to the modified DEM groups of 2 and 4 cycles.

[0027] 1.2 Preparation of modified DEM decellularized tracheal matrix

[0028] Three groups of New Zealand rabbits were sacrificed by injecting air into ear margin veins, and the trachea was isolated under the principle of asepsis, and the connective tissue on the outer wall of the trachea was immediately peeled off. Soak the fresh trachea of ​​group A (control group, n=10) in 4°C PBS buffer solution containing 1% antibiotics and antifungal drugs (AA) for testing. The other two groups of fresh trachea were divided into groups B and C.

[0029] First osmotically dissolve in sterile distilled water at 4°C for 48h, then enter the first decellularization cycle: soak in 4% w / v (g / ml) sodium de...

Embodiment 2

[0030] Embodiment 2 effect verification

[0031] method:

[0032] 1. Macroscopic measurement and biomechanical testing

[0033] Observe the general shape of the tracheal matrix of each group under direct vision, such as whether there is a defect or collapse; measure the length, thickness and width of the middle part of the trachea of ​​each group of samples, and test the anti-pressure properties of each group of samples with a universal mechanical testing machine. The fixed pressure plate sensor of the universal testing machine is used to fix the sample in the center of the disc, the initial load pressure is 0.1N, and the constant compression is started at a rate of 5mm / min at room temperature, and the time when the pipe diameter is compressed to 50% is recorded. Strain pressure, elastic modulus value, and recovery of deformation after compression.

[0034] 2. Routine histological staining analysis

[0035] The samples of each group were immersed in 10% neutral formaldehyde...

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Abstract

The invention provides a method for preparing a rabbit acellular tracheal matrix. The method includes the following steps: using a fresh rabbit trachea as a raw material, permeating and dissolving insterile distilled water at 4 DEG C for 48 hours, entering a first decellularization cycle: soaking in 4% w / v (g / ml) of a sodium deoxycholate solution, and placing in shaking table at 20 DEG C for performing incubation for 4 h; rinsing with sterile PBS for twice, then placing in 1 mol / L NaCl containing Dnase I at a concentration of 50 kU / mL, continuously shaking for 3 h to dissolve a nucleus and degrade DNA, and finally rinsing twice with the PBS; and performing a next decellularization cycle in the same procedure, and totally performing treatment of two cycles. The method unexpectedly discovers that through improved DEM treatment, it is only required to prepare an acellular matrix similar in structure of an original trachea in the two cycles, mucosal epithelial cells are completely removed, a few chondrocytes remain, a structure is complete, the immunogenicity is weak, infiltration of inflammatory cells is less, the clinical preparation time is shortened, and a treatment basis and hopeare provided for patients who need emergency tracheal stents.

Description

technical field [0001] The invention relates to a method for preparing rabbit decellularized tracheal matrix, which belongs to the technical field of bioengineering. Background technique [0002] Tracheal injury is mainly caused by tumor, infection, trauma, etc. When the length of the injury is less than 1 / 2 of the adult's full length or 1 / 3 of the child's, stump anastomosis is considered to be the most effective treatment for tracheal repair. When these malignant or benign lesions involve this length of the trachea, conventional surgical treatment is difficult to achieve good results. In recent years, with the rapid development of tissue engineering and the continuous deepening of research on tracheal transplant substitutes, tissue engineered trachea has gradually become one of the best methods of replacement therapy for patients with end-stage tracheal lesions. The best tracheal substitute should have similar anatomical structure and chemical and biological signaling syst...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3687A61L27/3691A61L27/3679A61L27/50A61L2430/40A61L2430/22
Inventor 王志豪史宏灿卢丹潘枢潘子寅
Owner YANGZHOU UNIV
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