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Acellular xenograft and preparation method thereof

A technology of nerve transplantation and decellularization, which is applied in the field of decellularized xenogeneic nerve grafts and its preparation, can solve the problems of small immune rejection, high price, and difficulty in obtaining materials, and achieve safe use, no immune rejection, and biophase good capacitive effect

Inactive Publication Date: 2016-11-23
YANTAI ZHENGHAI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgical treatment is aimed at long-term peripheral nerve injury, including direct surgical suture and nerve transplantation techniques, among which nerve transplantation techniques include autologous transplantation and nerve repair materials, etc. The former is because it can provide nerve growth factors and has less immune rejection. It is regarded as the gold standard for peripheral nerve repair, but it has the disadvantages of limited sources, new wounds, and complications such as donor site numbness, scars, and neuromas
Nerve repair materials currently used in clinical practice, such as allogeneic nerves, are mostly obtained from residual limbs, and are decellularized to ensure safety, no immune rejection, and no virus, but there are problems such as difficulty in obtaining materials and high prices.

Method used

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  • Acellular xenograft and preparation method thereof

Examples

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Embodiment 1

[0035] Embodiment 1, preparation of porcine decellularized xenograft

[0036] 1. Pre-treatment: The healthy adult pigs that have been slaughtered in the slaughterhouse are aseptically taken from the sciatic nerve, and fully washed with purified water.

[0037] 2. Alkali treatment: 1.5M NH 3 ·H 2 O aqueous solution, immerse at 4°C for 60 minutes, and wash thoroughly with purified water. To remove non-collagenous proteins in cell membranes.

[0038] 3. Surfactant treatment: firstly carry out SB-10 treatment, soak in 125mmol / L SB-10 at 4°C for 12 hours, wash fully with PBS; then use 0.14% TritonX-200, 0.6mmol / L SB-10 mixed solution Treatment, immersion treatment at 4°C for 7 hours. To destroy the phospholipid bilayer structure of the cell membrane, the cell ruptures, and the cytoplasm and nuclear substances are released.

[0039] 4. Degreasing treatment: place the product in isopropanol at 20°C and shake and soak for 8 hours. Remove the fat from the nerves.

[0040] 5. PBS...

Embodiment 2

[0042] Example 2: Preparation of bovine decellularized xenografts

[0043] 1. Pre-treatment: The healthy adult cattle that have been slaughtered in the slaughterhouse are aseptically taken from the ulnar nerve, and fully washed with purified water.

[0044] 2. Alkali treatment: 2M KOH aqueous solution, soak at 10°C for 100 minutes, and fully wash with purified water. To remove non-collagenous proteins in cell membranes.

[0045] 3. Surfactant treatment: firstly carry out SB-10 treatment, soak in 125mmol / L SB-10 at 10°C for 12 hours, wash fully with PBS; then use 0.14% TritonX-200, 0.6mmol / L SB-10 mixed solution Treatment, immersion treatment at 10°C for 7 hours. To destroy the phospholipid bilayer structure of the cell membrane, the cell ruptures, and the cytoplasm and nuclear substances are released.

[0046] 4. Degreasing treatment: place the product in acetone at 25°C and shake and soak for 10 hours. Remove the fat from the nerves.

[0047] 5. PBS buffer treatment: PBS...

Embodiment 3

[0050]Example 3: Preparation of canine decellularized nerve xenografts

[0051] 1. Pretreatment: Aseptically remove the radial nerve from healthy adult dogs that have been slaughtered in the slaughterhouse, and wash them thoroughly with purified water.

[0052] 2. Alkali treatment: 0.5M Ca(OH) 2 Aqueous solution, 20 ℃ immersion treatment for 40 minutes, fully washed with purified water. To remove non-collagenous proteins in cell membranes.

[0053] 3. Surfactant treatment: firstly carry out SB-10 treatment, soak in 125mmol / L SB-10 at 20°C for 12 hours, wash fully with PBS; then use 0.14% TritonX-200, 0.6mmol / L SB-10 mixed solution Processing, soaking at 20°C for 7 hours. To destroy the phospholipid bilayer structure of the cell membrane, the cell ruptures, and the cytoplasm and nuclear substances are released.

[0054] 4. Degreasing treatment: place the product in chloroform at 10°C and shake and soak for 9 hours. Remove the fat from the nerves. Remove the fat from the n...

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Abstract

The invention discloses an acellular xenograft and a preparation method thereof. The method includes the following steps: immersing peripheral nerves of the isolated mammal with alkaline solution, surfactant solution, degreasing agent and PBS buffer solution in order, freeze-drying and sterilizing to obtain the acellular xenograft. The acellular xenograft prepared by the invention has good histocompatibility, can regenerate nerve regeneration pathways, guide axon regeneration, accelerate nerve functional repair, and has good clinical application prospect in the field of repair of the peripheral nerves.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and in particular relates to a decellularized xenogeneic nerve graft and a preparation method thereof. Background technique [0002] Peripheral nerve injury is a common clinical trauma disease. With the development of society and the frequent occurrence of natural disasters in recent years, the number of nerve injuries caused by accidents, tumors, wars, earthquakes, etc. has increased year by year. According to statistics, more than one million people around the world will suffer from peripheral nerve injuries every year and cause Different degrees of functional impairment, loss, deformity and even disability have brought great pain to patients. Due to the complex pathological process after peripheral nerve injury, the inability of mature neurons to divide and replicate, and slow nerve repair and regeneration, how to effectively treat peripheral nerve injury has always been a difficult proble...

Claims

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Application Information

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IPC IPC(8): A61L27/36
CPCA61L27/3633A61L27/3675A61L27/3687A61L27/3691A61L2430/32A61L2430/40
Inventor 尹晓彤窦源东孙先昌董佳桓闫玉芳
Owner YANTAI ZHENGHAI BIO TECH
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