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Method for preparing tissue-engineered bone/cartilage integrated scaffold

A technology for tissue engineering bone and cartilage, applied in the field of tissue engineering technology and biomaterials, can solve the problems of complex process operation, poor shear resistance, obvious scaffold delamination, etc. Simple process effect

Inactive Publication Date: 2012-07-11
BEIJING SHENYOU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the scaffolds prepared by the above methods and materials have the following problems: (1) the internal layer of the scaffold is obvious, and the shear resistance between layers is poor; (2) the pore structure of the scaffold cartilage layer is low in bionicity, and there is no gradient orientation change (3) There are defects in the immediate fixation of the bracket and the repair site, which cannot meet the needs of clinical repair; (4) The mechanical support strength of gel materials is low, and the degradation of polymer materials produces acidic substances, which destroys the microenvironment of cartilage repair; ( 5) Part of the process is complex and requires expensive equipment

Method used

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  • Method for preparing tissue-engineered bone/cartilage integrated scaffold
  • Method for preparing tissue-engineered bone/cartilage integrated scaffold
  • Method for preparing tissue-engineered bone/cartilage integrated scaffold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Preparation of different concentrations of articular cartilage extracellular matrix (ECM)

[0037] Purchase fresh porcine articular cartilage tissue pieces, rinse them repeatedly with normal saline, cut them into small pieces, and put them in Tris-HCL (pH7.4) buffer containing protease inhibitor phenylmethylsulfonyl fluoride (0.35mM) , Ultrafine wet pulverization at 4°C. Add triple distilled water to fully dilute and centrifuge at 4°C for 30 minutes at a differential speed (3000-6000r / min), collect the supernatant, and use 1% TritonX-100 at 4°C for 24 hours to descaling and remove residual cell components. Add deoxyribonuclease 50000U / L and ribonuclease 1000U / L, and digest at 37°C for 12 hours to remove nuclear substances. Add 10-20ml of 75% ethanol solution for precipitation, centrifuge at 12000r / min at 4°C to collect the precipitate, add triple distilled water to prepare two kinds of slurries with a concentration of 1% and 4%, and store at 4°C for later us...

Embodiment 2

[0038] Embodiment 2: Preparation of bone matrix gelatin (BMG)

[0039] Select the cancellous bone of the Achyranthes knee joint, cut the cancellous bone into 1mm-thick bone slices with a non-decalcified bone slicer, soak and rinse with normal saline repeatedly for 15-30 minutes, then soak in 1% hydrogen peroxide for 1 hour to The bone block is milky white. After rinsing the bone blocks twice with triple distilled water, they were frozen together at -20°C for 24 hours and then thawed, and the cell membrane was broken by repeated freezing and thawing twice in hypotonicity. Add fat-free and decellularized solution: 2% sodium dodecyl sulfate, 6% lipase, 1% ethylenediaminetetraacetic acid, 0.1% sodium azide, shake at room temperature for 24 hours. Then the bone slices were put into 10% ethylenediaminetetraacetic acid solution and stored at 37° C. for 4 days until the bone slices took on the shape of porous white sponge. Then rinse the bone slices with 3% TritonX-100 for 12 hours,...

Embodiment 3

[0040] Example 3: Preparation of bone matrix gelatin and extracellular matrix integrated scaffold

[0041] Choose a mold with one end open and the other end as a piston-type movable bottom. First put the BMG sheet prepared in Example 2 into the bottom of the mold, get 20ml of the 4% ECM slurry prepared in Example 1 and pour it into the mold, vibrate for 2 minutes, so that the ECM slurry is completely soaked into the BMG pores, and the excess slurry is covered in the mold. The upper surface of the BMG has a thickness of 2mm; the concentration in Example 1 is 10ml of ECM slurry and continues to be added to the mold, covered with 4% ECM slurry, and the vertical vibration makes the two layers of ECM slurry blend naturally to form an integrated structure. The overall thickness is 3mm; adjust the depth of the mold so that the slurry is flush with the open end, cover the open end with a stainless steel sheet and fully contact the slurry, and wrap the outer periphery and bottom of the...

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Abstract

The invention discloses a method for preparing a tissue-engineered bone / cartilage integrated scaffold. An integrated scaffold structure of which internal pores are aligned and pore diameters are changed in a gradient way along a thickness direction and which has good mechanical properties is obtained by simulating the composition and spatial structure characteristics of natural articular cartilage and using extracellular matrixes and bone matrix gelatin of the cartilage as main raw materials by methods such as a different concentration superposition method, a directional crystallization method, a freeze-drying method, a cross-linking reaction method and the like. The high biocompatibility of the scaffold is favorable for the adhesion and proliferation of cells; a special pore structure of the scaffold ensures that the content of matrixes secreted by the cells is also changed in a gradient way; and meanwhile, a bone matrix gelatin substrate provides good mechanical support for the instant fixation of the scaffold and an injured part.

Description

technical field [0001] The invention belongs to the field of tissue engineering technology and biomaterials, relates to the technical field of preparation of tissue engineering scaffolds for repairing human articular cartilage defects, in particular to a tissue engineering bone / cartilage integrated scaffold with gradient function bionics and a preparation method thereof. Background technique [0002] Articular cartilage damage caused by factors such as trauma, degeneration, or inflammation is a common clinical disease in orthopedics. Due to the extremely limited ability of cartilage regeneration and repair, it is difficult to repair itself once damaged. Traditional treatment methods such as conservative treatment, microfracture, and arthroscopy Lavage, etc., cannot achieve satisfactory curative effect, and the continuous development of the injury site can lead to limb dysfunction and disability, which seriously affects the quality of life of patients. At present, autologous ...

Claims

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Application Information

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IPC IPC(8): A61L27/46A61L27/36A61L27/22A61L27/20A61L27/12A61L27/56C08J9/42C08J3/28C08J3/24
Inventor 梁增义
Owner BEIJING SHENYOU BIO TECH
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