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Preparation method of natural tissue-derived epiphyseal cartilage combined bone acellular material

A technology of epiphyseal cartilage and decellularization, which is applied in the field of preparation of epiphyseal cartilage combined with bone decellularized materials, can solve problems that have not been reported, and achieve good biocompatibility, considerable effect, and excellent cartilage combined with bone defect repair effect

Inactive Publication Date: 2021-11-02
THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on biomaterials that can be used to repair cartilage combined with bone, but there is no report on the repair of osteoarthritis and cartilage defects with epiphyseal cartilage combined with bone

Method used

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  • Preparation method of natural tissue-derived epiphyseal cartilage combined bone acellular material
  • Preparation method of natural tissue-derived epiphyseal cartilage combined bone acellular material
  • Preparation method of natural tissue-derived epiphyseal cartilage combined bone acellular material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A method for preparing epiphyseal cartilage combined with bone decellularized material derived from natural tissue, the specific steps are as follows:

[0051] (1) Material collection: take fresh 3-month-old juvenile Large White pig femoral distal epiphysis cartilage joint bone, rinse with sterile PBS buffer 5 times at 4°C for 10 minutes each time, and remove surface hair, muscle, and fascia , blood and interstitial fluid and other impurities; the size of the material is determined according to the actual needs, usually a cylinder with a diameter of 5mm and a height of 5mm;

[0052] (2) Decellularization process:

[0053] ①. In 500ml of organic solvent solution (20% acetone solution), place in a constant temperature shaker with a constant temperature of 25°C and a rotation speed of 60rpm for 4 hours to shake and degrease, and then rinse with sterile PBS for 12 hours;

[0054] ②. After drying the material with sterile gauze, place it in liquid nitrogen for 30 minutes, t...

Embodiment 2

[0076](1) Material collection: take fresh 3-month-old juvenile Large White pig femoral distal epiphysis cartilage joint bone, rinse with sterile PBS buffer 5 times at 4°C for 10 minutes each time, and remove surface hair, muscle, and fascia , blood and interstitial fluid and other impurities; the size of the material is determined according to the actual needs, usually a cylinder with a diameter of 5mm and a height of 5mm;

[0077] (2) Decellularization process:

[0078] ①. In 500ml of organic solvent solution (20% acetone solution), place in a constant temperature shaker with a constant temperature of 25°C and a rotation speed of 60rpm for 2 hours to shake and degrease, and then rinse with sterile PBS for 12 hours;

[0079] ②. After drying the material with sterile gauze, place it in liquid nitrogen for 30 minutes, then quickly take it out and place it in a water bath at 37°C for 30 minutes; repeat 3 times; finally rinse with sterile PBS twice, 5 minutes each time;

[0080] ...

Embodiment 3

[0087] (1) Material collection: take fresh 3-month-old juvenile Large White pig femoral distal epiphysis cartilage joint bone, rinse with sterile PBS buffer 5 times at 4°C for 10 minutes each time, and remove surface hair, muscle, and fascia , blood and interstitial fluid and other impurities; the size of the material is determined according to the actual needs, usually a cylinder with a diameter of 5mm and a height of 5mm;

[0088] (2) Decellularization process:

[0089] ①. In 500ml of organic solvent solution (30% acetone solution), place in a constant temperature shaker with a constant temperature of 25°C and a rotation speed of 60rpm to vibrate and degrease for 6 hours, and then rinse with sterile PBS for 12 hours;

[0090] ②. After drying the material with sterile gauze, place it in liquid nitrogen for 30 minutes, then quickly take it out and place it in a water bath at 37°C for 30 minutes; repeat 3 times; finally rinse with sterile PBS twice, 5 minutes each time;

[009...

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Abstract

The invention discloses a preparation method of a natural tissue-derived epiphyseal cartilage combined bone acellular material, and belongs to the technical field of biological materials for tissue or organ repair and regeneration. The preparation method comprises the following steps: rinsing and repeatedly freezing and thawing epiphyseal cartilage combined bone at the distal femur of a young large white pig, and treating the epiphyseal cartilage combined bone with a PBS buffer solution containing TritonX-100, a PBS buffer solution containing SLES, a PBS buffer solution containing HTHOPS, a PBS buffer solution containing DNaseI and normal saline to obtain the decellularized epiphyseal cartilage combined bone material. After thorough decellularization treatment is carried out on the epiphyseal and cartilage combined bone material, the completeness of the epiphyseal and cartilage combined bone ECM is reserved to the maximum extent while cells with immunogenicity and cell content components are completely removed, and the epiphyseal and cartilage combined bone ECM has the capability of repairing osteochondral defects and can be used for treating orthopedic diseases caused by osteochondral defects.

Description

technical field [0001] The invention relates to the technical field of biomaterials used for tissue or organ repair and regeneration, and more specifically relates to a preparation method of epiphyseal cartilage combined with bone decellularized material derived from natural tissues. Background technique [0002] At present, as China gradually enters an aging society, diseases such as osteoarthritis and cartilage damage and cartilage defects caused by strenuous activities are increasingly affecting human health, causing heavy economic and physical and mental burdens on individuals, and causing heavy social burdens. Problems that need to be solved urgently. In view of the above problems, the current traditional solutions include arthroscopic repair surgery, bone and joint replacement, etc. There are still many problems, such as poor repair effect and huge surgical trauma. In recent years, the use of biomaterials to repair cartilage tissue defects has received widespread atte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3612A61L27/3608A61L27/3654A61L27/3687A61L27/3691A61L27/50A61L2430/06A61L2430/24
Inventor 林贤丰张琪范顺武
Owner THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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