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Preparation and application of nerve tissue matrix derived tissue engineering scaffold material

A technology of tissue engineering scaffold and nerve tissue, which is applied in medical science, catheters, prostheses, etc., can solve the problems of inconsistent composition, poor biocompatibility, tube wall collapse, etc., and achieve good biocompatibility, no Effect of immune rejection, uniform distribution of pores

Inactive Publication Date: 2011-10-19
卢世璧
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Natural bioactive materials such as veins and skeletal muscle have problems such as tube wall collapse, poor regeneration, scar tissue hyperplasia and adhesion, and the sources are relatively limited, not suitable for mass production, and have certain immunogenicity
Artificially synthesized non-degradable materials still have many problems in terms of mechanical properties, degradation and absorption speed, and the negative impact of degraded acidic substances on nerve regeneration. Insufficiency such as inflammatory reaction in the product

Method used

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  • Preparation and application of nerve tissue matrix derived tissue engineering scaffold material
  • Preparation and application of nerve tissue matrix derived tissue engineering scaffold material
  • Preparation and application of nerve tissue matrix derived tissue engineering scaffold material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Preparation of nerve matrix-derived scaffold material

[0053] Take the canine sciatic nerve, remove the muscle and fat tissue around the nerve, rinse with sterile distilled water 2-3 times, 3-5 minutes each time; add to the phosphate buffer containing protease inhibitors 0.005-0.1% PMSF and 0.01-1% EDTA , crushed into small pieces by a pulverizer; expand the tissue with sodium citrate buffer (pH3-5), shake continuously at 4°C for 24-72 hours; wash 2-3 times with sterile distilled water, 3-5 times each time minutes; add 0.5-2% Triton X-100 in 10mM / L Tris-HCL buffer (pH7.5) at 4°C, which also contains protease inhibitors 0.005-0.1% PMSF and 0.01-1% EDTA, shake continuously for 24-72 hours to remove cell components; wash repeatedly with PBS after centrifugation; digest with DNase and RNase at 37°C for 6-48 hours; add 2M urea solution to dissolve for 48-72 hours, then mechanically pulverize again; centrifuge at 6000rpm for 20 minutes The undissolved substances ...

Embodiment 2

[0056] Example 2: Preparation of three-dimensional porous orientation scaffold

[0057] Use dilute acetic acid with pH=3.2 as the solvent to make the neuromatrix-derived powder into a suspension with a concentration of 3%, stir well, and centrifuge to remove air bubbles, then inject the centrifuged slurry into a mold, and place it vertically in a -40°C Pre-freeze on a frozen metal plate, store at -40°C for at least 24 hours after freezing completely, and then transfer to a freeze dryer for sublimation drying. The ice phase was sublimated under vacuum (60 After disinfection, seal and store at 4°C.

[0058] Physical and chemical performance testing: under an optical microscope, it can be seen that the stent pore size is uniform and the pores are interconnected (see Figure 7 ). Observation by scanning electron microscope shows that the stent pore size and distribution are relatively uniform, and the gaps are interconnected, and the pore size is 200-300 microns.

Embodiment 3

[0059] Example 3: Preparation of Nerve Regeneration Conduit

[0060] The neuromatrix-derived powder obtained in Example 1 was freeze-dried and dissolved in hexafluoroisopropanol (HFP), stirred thoroughly to prepare the solution, vacuumed to remove air bubbles, and then added to the syringe of the electrospinning instrument. Adjust the voltage of the electrospinning equipment, the distance between the needle and the receiving device, and the flow rate of the solution to prepare microfilaments with uniform thickness to form a thin film. Depending on the receiving device of the electrospinning equipment, a film with a certain direction or disorder can be prepared. The prepared electrospun film is taken out from the mould, and the distance from the light source is 5-10 cm, the wavelength is 258 nm, and the ultraviolet rays are cross-linked for 2-8 hours; For: EDC 50mM; NHS 20mM) After cross-linking at 4°C for 24h, rinse with three distilled water for 6 times, and freeze-dry. Wra...

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Abstract

The invention discloses preparation and application of a nerve tissue matrix derived tissue engineering scaffold material. A nerve tissue is taken as a raw material, matrix components (including nano-scale collagen microfilaments, fibronectin microfilaments and laminin microfilaments) favorable for nerve regeneration are extracted by means of medicine expansion, mechanical pulverization, enzymolysis treatment, dialysis collection and the like, and immunogenicity components (including Schwann cells, phospholipid and axons) not favorable for nerve regeneration are removed. The prepared nerve tissue matrix derived material can be further prepared into a three-dimensional porous oriented scaffold through oriented crystallization, freeze drying and crosslinking separately or by matching other polymer materials, or is prepared into a nano-scale film by an electrospinning technology, and the film is wound to form a nerve regeneration catheter. The tissue engineering scaffold or catheter prepared by the method is favorable for adhesion, proliferation and migration of seed cells, promotes nerve generation and can be used for never defect repair.

Description

technical field [0001] The invention belongs to the technical field of preparation of tissue engineering materials, and relates to the preparation technology of peripheral nerve tissue engineering bio-scaffold materials, in particular to a preparation method of a peripheral nerve matrix material for nerve tissue engineering that guides the regeneration of peripheral nerve tissue, and the use of the peripheral nerve matrix material A method for preparing a three-dimensional porous oriented scaffold and a nerve regeneration catheter and corresponding scaffold products. Background technique [0002] The repair and reconstruction of peripheral nerve defect is a major problem in the field of peripheral nerve injury. At present, autologous nerve transplantation is still the standard operation for solving nerve defects in the clinical treatment process. However, due to the limited source of autologous nerves and many complications in the donor site, the medical community has been l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L29/00A61L29/04
Inventor 卢世璧王玉彭江赵喆黄靖香张莉许文静
Owner 卢世璧
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