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A kind of decellularized nerve matrix material and its preparation method and application

A matrix material and decellularization technology, applied in the field of decellularized neural matrix materials and its preparation, can solve the problems of poor biocompatibility, high cost, and lack of biological activity

Active Publication Date: 2021-05-14
SHANDONG EYE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, biological tissues generally have an extremely complex ultrastructure, and it is extremely difficult to synthesize materials close to the ultrastructure of organisms by artificial methods; in addition, the cost of synthetic organic polymer materials is generally high, and the materials themselves do not have biological activity. The compatibility is poor, and it cannot trigger specific biological reactions. It is necessary to surface treat the material and add biological factors to improve the adhesion of cells to the matrix material; and the degradation products of the material may trigger the body's inflammatory response

Method used

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  • A kind of decellularized nerve matrix material and its preparation method and application
  • A kind of decellularized nerve matrix material and its preparation method and application
  • A kind of decellularized nerve matrix material and its preparation method and application

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preparation example Construction

[0031] The invention provides a preparation method of decellularized nerve matrix material, comprising the following steps:

[0032] (1) Disinfect the nerve tissue block and wash it with a sterile buffer solution to obtain the sterilized nerve tissue block;

[0033] (2) Shaking and crushing the sterilized nerve tissue block in sterile water to obtain the crushed nerve tissue block;

[0034] (3) Enzymolyzing the crushed nerve tissue block in a sterile buffer solution containing nuclease to obtain a nucleic acid-depleted nerve tissue block;

[0035] (4) Soak the nucleic acid-depleted nerve tissue block in a sterile buffer solution containing 0.5-5% TrionX-100 by volume to obtain a cell-membraned nerve tissue block;

[0036] (5) enzymatically hydrolyzing the nerve tissue blocks with decellularized membranes in a sterile buffer containing chondroitinase to obtain chondroitin-desulfated nerve tissue blocks;

[0037] (6) Put the chondroitin-desulfated nerve tissue block in 0.2-1 m...

Embodiment 1

[0052] Preparation of Decellularized Matrix of Porcine Peripheral Nerve

[0053] (1) Material collection: the tibial nerve of a healthy pig was selected, washed twice with PBS to remove blood stains. As for the EP tube, store it in a -80°C refrigerator for later use.

[0054] (2) Disinfection and preparation: Soak the tissue block in 75% alcohol for 2-3 seconds, wash it twice with PBS, and place it in a 250ml jar.

[0055] (3) Pre-cleaning and crushing: Add 250ml of sterile third-grade water, place on a vibrating shaker, set at 200rpm / min, and treat at 4°C for 12 hours.

[0056] (4) Residual nucleic acid removal: transfer the nerve tissue to an EP tube containing 1000 units of nuclease in 1 ml of sterile D-Hank's buffer, and place it on a vibrating shaker at 100 rpm / min, and treat at 37°C for 12 hours.

[0057] (5) Broken and cleaned: the nervous tissue was transferred to a jar filled with 250 ml of 1% TrionX-100 sterile PBS solution, placed on a vibrating shaker, set at 200...

Embodiment 2

[0067] Biocompatibility of acellular neural matrix materials

[0068] (1) Choose healthy male New Zealand white rabbits with a weight of 3-3.5KG, and remove 2cm of the facial nerve on the right cheek of the rabbit;

[0069] (2) Cut the decellularized nerve matrix material to a length of 2 cm, wash it with normal saline, and suture it at the nerve defect site;

[0070] (3) Suture the surgical incision;

[0071] (4) After two weeks, the sutures were removed, and the nerve repair and the degree of inflammatory response were detected.

[0072] The rabbits transplanted with the acellular neural matrix material showed a healthy living condition within two weeks, no complications occurred, and there was no abnormality in chewing and hearing. Transplant results such as image 3 as shown, image 3 -a is the photo of decellularized neural matrix material; image 3-b is the anatomical picture two weeks after the transplantation of the acellular neural matrix material, in which the t...

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Abstract

The invention discloses a preparation method of an acellular nerve matrix material, which belongs to the field of biomedical material processing. The method of the present invention sterilizes, cleans, shakes and breaks the nerve tissue block, enzymatically hydrolyzes with nuclease, soaks in TrionX-100, enzymolyzes with chondroitin sulfate enzyme, soaks and cleans with NaCl, and finally obtains a compound with good biocompatibility and suitable biological properties. Degradation rates of neural decellularized matrix materials. The material can be used for nerve regeneration repair. At the same time, the raw materials of the decellularized neural matrix material in the present invention have wide sources, are readily available, are cheap, and are easy to manufacture.

Description

technical field [0001] The invention belongs to the field of biomedical material processing, and in particular relates to a decellularized nerve matrix material and its preparation method and application. Background technique [0002] The repair and treatment of peripheral nerve defects has always been a difficult problem in clinical work, especially for long-segment nerve defects larger than 2-3cm. Shorter peripheral nerve defects can be sutured directly, while autologous nerve tissue grafts are considered to be a good repair material for long-segment nerve defects. However, since the nerves for transplantation are cutaneous nerves, it is difficult to meet the needs of clinical nerve transplantation, and it will cause permanent nerve dysfunction in the donor area. [0003] The development of tissue engineering provides a new solution for the repair of nerve defects. At present, there are many applications such as artificially synthesized polymer materials and decellulariz...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/58A61L31/00A61L31/14A61K35/30A61P25/00
CPCA61K35/30A61L27/3604A61L27/3675A61L27/3687A61L27/3691A61L27/58A61L31/005A61L31/148A61L2300/412A61L2430/32A61L2430/40A61P25/00
Inventor 周庆军赵龙史伟云
Owner SHANDONG EYE INST
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