Tissue engineering cartilage and preparation method thereof

A technology of tissue engineering and cartilage, applied in bone implants, medical science, prostheses, etc., can solve the problems of limited donor sources, failure to meet normal cartilage mechanical requirements, increase patient damage, etc., and achieve good cell activity and micro-organisms. Effects of environment, improvement of cartilage survival rate, and improvement of fusion ability

Active Publication Date: 2012-07-04
SHAANXI BIO REGENERATIVE MEDICINE CO LTD
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AI Technical Summary

Problems solved by technology

For example, joint debridement can only temporarily relieve symptoms, but cannot effectively treat cartilage damage; microfracture surgery can only regenerate fibrocartilage at the defect site, which cannot meet the mechanical requirements of normal cartilage, and there is a hidden danger of later degeneration; autologous cartilage transplantation is better, Only suitable for small cartilage defects (2 ) injury treatment; allogeneic cartilage transplantation can be used to repair larger defects, but the source of donors is limited, and there are risks of infectious diseases and immune rejection
Clinical studies have also shown that chondrocyte transplantation is currently an effective way to repair articular cartilage damage and achieve functional reconstruction; the first-generation transplantation technology needs to use the patient's autologous periosteum to seal the defect position, which increases the damage to the patient and is prone to periosteum hypertrophy. Second

Method used

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  • Tissue engineering cartilage and preparation method thereof
  • Tissue engineering cartilage and preparation method thereof
  • Tissue engineering cartilage and preparation method thereof

Examples

Experimental program
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Effect test

example 1

[0033] Example 1, preparation of human tissue engineered cartilage

[0034] Step 1. Preparation of cartilage calcification layer: Human bone marrow mesenchymal stem cells were amplified with commercial low-sugar DMEM medium, and the third generation BMSCs were taken at 1×10 5 / cm 2 Inoculated in a 6-well culture plate for cell culture, after the cells were completely adhered to the wall, the culture solution A was replaced to induce culture; Bovine serum 100ml / L, dexamethasone 10 -8 mol / L, β-sodium glycerophosphate 10mmol / L and ascorbic acid 50μg / mL.

[0035] Detection of induced differentiation into osteoblasts: Alkaline phosphatase calcium cobalt staining, calcium alizarin red staining and type I collagen immunohistochemistry were performed on the 7th and 14th days of culture to detect the induction of BMSCs into osteoblasts, and the results Positive; the preparation of cartilage calcification layer was completed after 15 days of culture.

[0036] The acquisition and cul...

example 2

[0044] Example 2, preparation of rabbit tissue engineered cartilage

[0045] Step 1. Preparation of cartilage calcification layer: take the third generation rabbit BMSCs according to the ratio of 1.5×10 5 / cm 2 Inoculate in a 12-well culture plate for cell culture at a density of 12, and replace the culture medium A after the cells adhere to the wall for induction culture; the composition of the culture medium A is: contained in commercial high-sugar DMEM culture medium, fetal bovine serum 100ml / L, dexamethasone 10 -8 mol / L, β-sodium glycerophosphate 10mmol / L and ascorbic acid 75μg / mL.

[0046] Detection of induced differentiation into osteoblasts: according to the detection method of Example 1, the result was positive; after 14 days of culture, the preparation of cartilage calcification layer was completed.

[0047] Step 2. Preparation of deep layer of cartilage: Take the third generation BMSCs and press 1×10 5 Inoculate in a 12-well culture plate for cell culture at a d...

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Abstract

The invention discloses a tissue engineering cartilage and a preparation method thereof. The tissue engineering cartilage is cytoactive, consists of sequentially laminated cell patches including a surface layer, a middle layer, a deep layer and a calcified layer, and takes collagen matrix as a main component; the layers are compounded by collagen and hyaluronic acid, and the bordering integration of various layers of cartilage structures is promoted through hydrostatic culture; and the tissue engineering cartilage is prepared by preparing various layers of cartilage cell patches and assembling multiple structural layers. The prepared tissue engineering cartilage is in a layering arrangement of a natural cartilage, has a very good tissue structure and very good mechanical properties (frictional coefficient is 75-85 percent that of a natural cartilage tissue, and the elastic modulus is 80-90 percent that of the natural cartilage tissue.); the integration between different layers is good; experiments prove that the prepared tissue engineering cartilage can form the tidal line same as that of a normal cartilage tissue and has the same layering structure; and the prepared tissue engineering cartilage is capable of promoting regeneration and function recovery of a cartilage and obviously shortening the cycle of cartilage damage treatment and is applicable to cartilage defect restoration for a load-bearing part of a articular cartilage.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering medical biomaterials, in particular to a tissue engineering cartilage with natural structure and function and a preparation method thereof. Background technique [0002] The composition and structure of natural articular cartilage, as the depth changes, the size, shape and density of chondrocytes also change. The content of collagen decreases with the increase of depth, while the content of proteoglycan increases with the increase of depth. According to the morphological changes of chondrocytes and cartilage interstitium, articular cartilage is divided into four layers from the articular surface to the bone, which are superficial layer, middle layer, deep layer and calcified layer. There is no clear boundary between the layers, but their composition, structure and mechanical properties are different, and the cell morphology and function are also different. The surface layer is the thin...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36A61L27/24A61L27/20A61F2/28
Inventor 田智泉
Owner SHAANXI BIO REGENERATIVE MEDICINE CO LTD
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