Composite structured tissue engineering cartilage graft and its preparation method

A composite structure and tissue engineering technology, which is applied in the fields of medicine and biomedical engineering, can solve the problems of biological glue not reaching the effective concentration, causing great damage to patients, and decreasing viscosity, and achieve the effects of saving chondrocytes, reducing pain, and not easy to fall off

Inactive Publication Date: 2006-06-14
北京市创伤骨科研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, cartilage defects often occur in joints (such as knee joints) where there is a lot of tissue fluid. Bio-protein glue is easily diluted or degraded too quickly by protease in tissue fluid, so that the bio-glue cannot reach the effective con

Method used

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  • Composite structured tissue engineering cartilage graft and its preparation method
  • Composite structured tissue engineering cartilage graft and its preparation method

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Experimental program
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Effect test

Embodiment 1

[0030] Example 1 Preparation and molding of allogeneic demineralized bone:

[0031] 1. The cancellous bone part of the allograft bone was made into a cylinder with a diameter of 4 mm and a length of 6 mm.

[0032] 2. Defat and protein the prepared cylindrical allogeneic bone to prepare antigen-free bone: use surfactant to defat and decellularize. For the specific method, refer to the patented method for the preparation of biologically derived artificial bone without antigen.

[0033] 3. Demineralize the allogeneic bone of the cylinder with hydrochloric acid at a concentration of 0.1-2.0 mol / L at a weight ratio of 50:1, and the demineralization time is 0.5-3 hours. In this embodiment, the concentration of hydrochloric acid is 0.5 mol / L, the weight ratio of hydrochloric acid to artificial bone is 50:1, and the demineralization time is 1 hour.

[0034] 4. After decellularization, degreasing and demineralization, the allograft bone forms an internally connected grid structure.

...

Embodiment 2

[0037] The acquisition of embodiment 2 chondrocytes:

[0038]We use dogs as experimental subjects. The knee joint of the dog's left hind leg was exposed, some cartilage in the non-load-bearing area of ​​the lower end of the femur was taken, and the joint cavity was sutured. Soak the removed cartilage tissue in sterile PBS (concentration 0.01mol / L, pH7.4±0.1) containing gentamicin (40 units / ml) at a ratio of 200mg cartilage / 50ml PBS for 45-60 minutes, Afterwards, the cartilage tissue was manually cut into small pieces of 1mm×1mm×1mm size; the crushed cartilage tissue pieces were placed in 0.2% mg / ml type II collagenase solution, and digested in a constant temperature oscillator at 37°C±0.5°C for 8- After 12 hours, filter and centrifuge; the precipitated cells were washed twice with sterile PBS (concentration 0.01 mol / L, pH 7.4±0.1), stained with trypan blue to detect cell viability, and the viability was > 60%, and set aside.

Embodiment 3

[0039] The preparation of embodiment 3 cartilage tissue engineering graft: (following steps all require aseptic operation)

[0040] 1. Put 2.5×10 5 chondrocytes with 25 μl containing thrombin and Ca 2+ The solution (sterilized) was mixed evenly, and prepared into solution 4 for later use.

[0041] 2. Take the solution whose main components are fibrinogen and factor XIII as solution 5, inhale solution 5 into another needle syringe, and the inhalation volume is equal to solution 4.

[0042] 3. Place the cylindrical solid carrier with a diameter of 4 mm and a length of 6 mm upright, and slowly inject solution 4 and solution 5 into the cylindrical solid carrier after the needle of the syringe containing solution 4 and solution 5 is pressed against the top of the cylindrical solid carrier.

[0043] 4. Let it stand for 5 minutes until the mixture of solution 4 and solution 5 coagulates. At this time, the colloidal carrier formed by the agglutination of the mixture of solution 4 a...

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Abstract

A tissue-engineered cartilage transplant with composite structure for treating the cartilage defect and surficial depressed defect is composed of the medically acceptable solid carrier with internal cavity, and the mixture of medically acceptable colloid and normal cartilage cells through filling said mixture in said cavity of biodegradable solid carrier.

Description

Technical field: [0001] The invention relates to the fields of medicine and biomedical engineering, in particular to a tissue engineered cartilage graft and a preparation method thereof. Background technique: [0002] Cartilage tissue is a single tissue containing only chondrocytes, without blood vessels inside, and its nutrition mainly depends on the penetration of interstitial fluid. It is a kind of connective tissue that has the functions of supporting, protecting, and dispersing stress. Histologically, it can be divided into three categories: hyaline cartilage (articular cartilage, costal cartilage), elastic cartilage (ear cartilage), and fibrocartilage (meniscus). Structurally, chondrocytes are located within the cartilage lacuna and are surrounded by the surrounding cartilage capsule. The cartilage capsule acts as a barrier separating the chondrocytes from the surrounding tissue. Therefore, cartilage tissue has the characteristics of weak antigenicity and mild immune...

Claims

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Application Information

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IPC IPC(8): A61L27/36
Inventor 孙磊田伟陈磊江健陶剑峰
Owner 北京市创伤骨科研究所
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