Tissue engineering nerve graft and application thereof

A tissue engineering and graft technology, applied in the fields of medicine and biomedical engineering, can solve the problem of difficulty in obtaining materials, and achieve the effects of abundant sources, easy separation and purification, and simple preparation process.

Inactive Publication Date: 2012-02-08
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the late start of research on neural stem cells, most of the established neural stem cell lines are derived from mice, and there are obvious species differences between mice and humans. If autologous cells are used, neural stem cells and glial cells are located in the central nervous system. The nervous system is very difficult to obtain

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Preparation of Chitosan Tissue Engineering Nerve from Autologous Bone Marrow Stem Cells

[0017] Chitosan was dissolved with 2% lactic acid to obtain a 1.5% chitosan solution. Then inject the solution into the mold, freeze and preform at -60°C, remove the material from the mold, put in 5% sodium hydroxide solution, neutralize the acid in the material, and after fixed molding, remove the excess alkali and the generated salt with distilled water After being cleaned, freeze-dried to obtain a chitosan artificial nerve graft containing neurotrophic factors, with a porosity of 87% and an average pore diameter of 220-289 μm.

[0018] Enrichment process of autologous bone marrow stem cells: extract about 40ml of subject's bone marrow, anticoagulate with heparin, pay attention to avoid clot formation as much as possible, transfer the bone marrow to two 50ml sterile centrifuge tubes in the ultra-clean bench, dilute them with PBS and Mix well, draw FICOLL10-20ml / tube×4 ...

Embodiment 2

[0021] Example 2 Chitosan Tissue Engineering Nerves Using Autologous Bone Marrow Mesenchymal Stem Cells (MSCs)

[0022] Chitosan was dissolved with 2% acetic acid to obtain a 2% solution of chitosan. Then inject the solution into the mold, freeze and preform at -70°C, remove the material from the mold, put it into a 10% sodium carbonate solution, neutralize the acid in the material and fix the molding, then remove the excess alkali and the generated salt with distilled water After cleaning, and then freeze-drying to obtain a chitosan artificial nerve graft, the porosity is 72%, and the average pore diameter is 130-180 μm.

[0023] Autologous bone marrow mesenchymal stem cell enrichment process:

[0024] (1) Cell separation

[0025] 1. Extract about 40ml of the subject's bone marrow, anticoagulate with heparin, and try to avoid clot formation

[0026] 2. Transfer the bone marrow to two 50ml sterile centrifuge tubes in the ultra-clean bench, dilute them with PBS and mix the...

Embodiment 3

[0076] Example 3 Chitosan Tissue Engineering Nerves Using Schwann Cells

[0077] Dissolve chitosan with 2% citric acid to obtain a 5% solution of chitosan, then pour the solution into the mold, freeze and preform at -80°C, and then remove the material from the mold and put it into 5% hydrogen In the sodium oxide solution, after neutralizing the acid in the material and fixing it into shape, after cleaning the excess alkali and the generated salt with distilled water, then freeze-drying to obtain the chitosan artificial nerve graft, the porosity is 51%, and the average pore size is 57-96 μm.

[0078] The chitosan artificial nerve guide prepared above was high-pressure sterilized for use. The day before the experiment, soak in DMEM+15%FBS complete medium for 24h. Dorsal root ganglia were obtained from embryonic 16d SD rats L4,5,6. Three DRGs were implanted at both ends of the prepared silk tube, cultured in DMEM+15%FBS in the bioreactor for 1 day, and then replaced with 97% n...

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Abstract

The invention discloses a chitosan artificial nerve graft containing a neurotrophic factor and a preparation method thereof. In the chitosan artificial nerve graft, chitosan is processed into a nerve conduit with a porous structure and high tensile strength by performing processes such as weak acid dissolving, injection molding, molding, neutralization fixing, cleaning, freeze drying and the like under the condition of not adding any foaming agent or crosslinking agent. The nerve conduit contains cells or tissues which have treating effects and are included or distributed inside or on the surfaces of tubular body pores, including autologous bone marrow stem cells, autologous bone mesenchymal stem cells, schwann cells or dorsal root ganglion tissues or combinations thereof. A tissue engineering nerve can be used for repairing peripheral nerve defect and can be applied to repairing of spinal cord injury simultaneously.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, in particular to tissue engineered nerve grafts containing cells or tissues and uses thereof. Background technique [0002] In the real world, accidents such as traffic accidents, industrial accidents, sports accidents, and clinical operations will cause peripheral nerve damage. Clinically, when long-distance nerve defects cannot be compensated for by end-to-end sutures, grafts have to be relied on. Come bridge fix. In recent years, tissue engineering based on the principles and technologies of life science and engineering has provided a new way to construct nerve transplant substitutes. The research on using tissue engineering technology to repair nerve damage mainly focuses on the design and construction of new tissue-engineered nerve bridge devices. At present, such bridges usually use biomaterials alone, and process them into a tubular structure of nerve guide channels to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/36A61F2/02
Inventor 顾晓松丁斐刘红顾剑辉汤欣杨宇民
Owner NANTONG UNIVERSITY
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