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Tissue-engineered peripheral nerve tissue and preparation method thereof

A technology of peripheral nerve and tissue engineering, applied in biochemical equipment and methods, tissue regeneration, tissue culture, etc., can solve the problems of insufficient cell volume, influence of cell biological activity, limited length and diameter of autologous nerves, etc.

Pending Publication Date: 2021-05-07
西安交通大学口腔医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, autologous nerve transplantation also has obvious disadvantages: (1) The length and diameter of the obtained autologous nerve are limited, which affects the efficacy of nerve repair due to the mismatch between part of the autologous nerve and the recipient nerve; (2) The autologous nerve cannot be harvested. Avoidance of surrounding tissue damage, deformity and loss of nerve function at the donor site due to surgery
A number of inventions are based on the development of nerve tissue combined with tissue engineering and regenerative medicine technology, and the strategy of seed cell / scaffold material complex for nerve tissue regeneration and repair has been designed. However, this strategy currently has some shortcomings: (1) Most of them use early ectodermal Mesenchymal stem cells or neuron cells are used as seed cells, which are difficult to obtain clinically and have insufficient cell volume; (2) Seed cells need to be supported, transferred and fixed by scaffold materials such as biological glue, which has a certain impact on the biological activity of cells
[0005] In summary, most of the tissue-engineered peripheral nerve tissues currently used for peripheral nerve regeneration can promote nerve tissue regeneration, but only a small amount of cells remaining at the stump of the nerve proliferate and form myelin, and it is difficult to obtain a good result due to incomplete myelination. Support and guide nerve regeneration

Method used

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  • Tissue-engineered peripheral nerve tissue and preparation method thereof
  • Tissue-engineered peripheral nerve tissue and preparation method thereof
  • Tissue-engineered peripheral nerve tissue and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The tissue-engineered peripheral nerve tissue prepared in this example uses dedifferentiated adipocytes as seed cells, which are separated, cultured, amplified, and induced into Schwann cells in vitro, and then dedifferentiated adipocyte aggregates are prepared, and finally the cell aggregates are mixed with The silk fibroin catheter scaffold material is compounded to form tissue engineered peripheral nerve tissue, and the specific steps of its preparation are as follows:

[0042] (1) Take 8 mL of adipose tissue from the oral buccal fat pad, thigh or abdominal subcutaneous fat tissue, put it in PBS containing penicillin and streptomycin 100 U / mL, soak it in a petri dish, and wash it 3 times for 4 minutes each time. Adipose tissue shredded to 1mm 3 , add 0.25% type I collagenase 3 times the volume of adipose tissue, shake, digest at 37°C for 1 hour, add stem cell culture medium to stop digestion. Filter with 200 μm and 100 μm filters in turn, and collect the upper layer...

Embodiment 2

[0052] The tissue-engineered peripheral nerve tissue prepared in this example uses bone marrow mesenchymal stem cells as seed cells, which are separated, cultured, expanded and induced into Schwann cells in vitro, and bone marrow mesenchymal stem cell aggregates are then prepared. Mesenchymal stem cell aggregates and scaffold materials are compounded in vitro to form tissue-engineered peripheral nerve tissue. The specific steps of preparation are as follows:

[0053] (1) Isolation, cultivation and screening of bone marrow mesenchymal stem cells: 3 mL was extracted from the ilium, injected into a centrifuge tube containing 0.3 mL heparin sodium saline (1000 U / mL), shaken and then added 6 mL of lymphocyte separation solution (1.2 g / mL), remove the white membranous layer of mononuclear cells after centrifugation, wash twice with PBS, discard the supernatant after centrifugation, add stem cell medium to resuspend, and place in 5% CO 2 , Cultured at 37°C, changing the medium every ...

Embodiment 3

[0063] The tissue-engineered peripheral nerve tissue prepared in this example uses adipose-derived stem cells as seed cells, which are separated, cultured, amplified, and induced into Schwann cells in vitro, and then adipose-derived stem cell aggregates are prepared. Composite to form tissue engineered peripheral nerve tissue, the specific steps of its preparation are as follows:

[0064] (1) Take 6 mL of adipose tissue from the oral buccal fat pad, thigh or abdominal subcutaneous fat tissue, put it in PBS containing penicillin and streptomycin 100 U / mL, soak it in a petri dish, and wash it 3 times for 3 minutes each time. Tissue minced to 1mm 3 , add 0.3% type I collagenase 3 times the volume of adipose tissue, shake, digest at 37°C for 1 hour, add stem cell medium to stop digestion. Use 200 μm and 100 μm filters to filter in turn, centrifuge to collect the lower layer of stromal vascular cell mixture (SVF), resuspend in PBS, filter with a 70 μm filter, discard the supernatant...

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Abstract

The invention discloses a tissue-engineered peripheral nerve tissue and a preparation method thereof, and belongs to the technical field of tissue engineering. The preparation method comprises the following steps of: separating, culturing, purifying and amplifying seed cells, inducing the seed cells in vitro to differentiate into Schwann cells, then preparing a stem cell polymer, and implanting the stem cell polymer into a silk fibroin catheter scaffold material to form the tissue-engineered peripheral nerve tissue. The tissue-engineered peripheral nerve tissue prepared by the method can be used for defect regeneration and repair of peripheral nerve tissues such as facial nerves and trigeminal branches. Compared with other types of tissue-engineered nerves existing at present, the tissue-engineered peripheral nerve tissue has the advantages that the seed cells are easy to obtain, the wound is small, the Schwann cells induced and differentiated in vitro are secreted into nerve-related factors to improve the nerve regeneration efficiency, the stem cell polymer contains sufficient cell quantity and rich extracellular matrix, and meanwhile, the tissue-engineered peripheral nerve tissue has better biological activity and good biological safety, and can significantly shorten the nerve repair time.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a tissue-engineered peripheral nerve tissue and a preparation method thereof. Background technique [0002] Destruction of nerve blocks due to trauma, tumor invasion, or surgery is a common clinical problem. Nerve tissue damage is accompanied by nerve dysfunction, such as facial expression muscle movement disorders caused by facial nerve branch damage, which will bring great physical and mental harm and pain to patients and their families. Therefore, exploring methods for repairing nerve tissue defects and regenerating biologically active nerve tissue has great clinical significance for the recovery of tissue function and the improvement of the quality of life of patients. [0003] At present, autologous nerve transplantation is the main method of clinically repairing peripheral nerve tissue defects, and this method has also achieved good results. However, autologous...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36A61L27/22A61L27/50A61L27/54C12N5/077C12N5/0775C12N5/079
CPCA61L27/3878A61L27/383A61L27/3834A61L27/3804A61L27/3687A61L27/227C12N5/0653C12N5/0667C12N5/0663C12N5/0622A61L27/50A61L27/54C12N2509/00C12N2509/10C12N2501/115C12N2501/135C12N2506/1384A61L2430/32C08L89/00
Inventor 那思家唐安群屠军波张舟崔浩
Owner 西安交通大学口腔医院
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