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Articular cartilage graft and preparation method thereof

A technology for articular cartilage and grafts, applied in joint implants, joint implants, medical science and other directions, can solve the problems of affecting the integration effect, poor biocompatibility, difficult cell migration, etc., and achieve the promotion of extracellular matrix. Secretion, good compression and abrasion resistance, improved long-term efficacy

Active Publication Date: 2014-07-16
西安博鸿生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] After retrieval, Chinese patents 02136560.1, 03147950.2, and 200310109203.6 disclose methods for constructing tissue engineered cartilage derived from allogeneic cells, but none of them carry out subregional construction according to the structure and function of normal cartilage, which may cause the prepared tissue engineered cartilage to be different from the surrounding normal cartilage and cartilage. The lower osseointegration is not good, and the wear resistance and compression resistance consistent with natural cartilage cannot be obtained. The patent also does not take effective measures to reduce the risk of immune rejection of allogeneic cells, and cannot guarantee the survival and function of cells in the recipient
Chinese patents 200610026862.7 and 200810102842.2 respectively use adipose-derived mesenchymal stem cells and umbilical cord mesenchymal stem cells to induce differentiation into chondrocytes as seed cells. Although mesenchymal stem cells come from a wide range of sources, mesenchymal stem cells induced in the direction of cartilage may be involved in gene expression, Extracellular matrix secretion, cell signal transduction, etc. cannot be exactly the same as normal chondrocytes, and it is uncertain whether mesenchymal stem cells induced in the direction of chondrocytes can maintain the characteristics of hyaline cartilage stably for a long time
In addition, when constructing tissue engineered cartilage, the biocompatibility of artificially synthesized scaffold materials is poor, and the degradation products may have adverse effects on the body
Without the use of scaffold materials, it is impossible to provide a three-dimensional space for the growth of chondrocytes, and it is impossible to solve the problems of chondrocyte aging and fibrosis during the culture process
Chinese patent 200710078264.9 constructs a bionic function interface osteochondral composite tissue integrated engineering scaffold with cartilage layer, calcification layer and subchondral bone layer, but the corresponding thickness of each layer is not set according to the thickness of normal cartilage, which is easy to cause The thickness of each layer of the constructed scaffold is inconsistent with that of the surrounding normal tissue during clinical application. After implantation, each layer of the constructed scaffold is mismatched with each layer of the surrounding normal cartilage, which affects the integration effect
In addition, because chondrocytes are surrounded by a large amount of extracellular matrix, cell migration is difficult. Simple scaffold materials used for cartilage tissue repair can only achieve fibrocartilage repair, and the long-term effect is not good.

Method used

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  • Articular cartilage graft and preparation method thereof
  • Articular cartilage graft and preparation method thereof
  • Articular cartilage graft and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Step 1. Obtain chondrocytes: cut the rabbit superficial cartilage tissue into 0.5~3mm 3 Size, first digest with 10 times volume of 0.1% (w / v) hyaluronidase solution for 30 minutes, shake once every 15 minutes, rinse with PBS twice after digestion; then add 10 times volume of 0.2% (w / v) ) Digestion with type II collagenase solution for 4 hours, stop the digestion with PBS and fully pipette the cells, filter with a 200-mesh sieve, and wash twice with PBS to isolate the primary superficial chondrocytes at a cell density of 1×10 5 Pcs / cm 2 Inoculate in culture flask, add culture solution A, 37℃, 5% CO 2 After culturing in the incubator for 3 days, when the superficial chondrocytes reach 80% confluence, they are subcultured. Add 10 times the volume of 0.25% (w / v) trypsin solution to digest for 3 minutes, stop with PBS and wash twice, according to the cell density of 1×10 4 Pcs / cm 2 Inoculate, add culture medium A, and subculture once every 3 days;

[0031] Cut the rabbit middle c...

Embodiment 2

[0043] Step 1. Obtain chondrocytes: cut the superficial layer of porcine cartilage into 0.5~3mm 3 Size, first digest with 10 times volume of 0.15% (w / v) hyaluronidase solution for 45 minutes, shake once every 15 minutes, rinse with PBS twice after digestion; then add 10 times volume of 0.25% (w / v) v) Digestion with type II collagenase solution for 8 hours, stop the digestion with PBS and fully pipette, filter with 200 mesh screen and wash twice with PBS to isolate primary superficial chondrocytes at a cell density of 5×10 5 Pcs / cm 2 Inoculate in culture flask, add culture solution A, 37℃, 5% CO 2 After 7 days of culture in the incubator, when chondrocytes reach more than 90% confluence, pass them down, digest with 10 times the volume of 0.5% (w / v) trypsin solution for 8 minutes, stop with PBS and wash twice, according to cell density 1×10 5 Pcs / cm 2 Inoculate, add culture medium A, passage once every 7 days, and change the medium every 3 days;

[0044] Cut the porcine middle cartil...

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Abstract

An articular cartilage graft and a preparation method thereof are provided. The prepared articular cartilage graft is composed of a superficial layer, a middle layer and a deep layer from outside to inside; the thickness, shape and size of each layer are all matched with a cartilage injury part, and thus preoperative shaping is not required; after grafting, the articular cartilage graft has layer distribution corresponding to distribution of each layer of surrounding normal cartilages, is conducive to intercellular signal transmission, transduction and regulation, and can be better integrated with surrounding normal cartilage tissues; the articular cartilage graft has structure characteristics consistent with those of the natural cartilages, the collagen type II content is gradually decreased from the superficial layer to the deep layer, the GAG content is increased gradually from the superficial layer to the deep layer, and compression resistance and wear resistance are good; and chondrocytes in the cartilage graft are wrapped with an extracellular matrix, have low immunogenicity, allow generation of immunologic rejection to be avoided after grafting, can survive for a long term and exert functions, and improve cartilage repair long-term curative effects.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering medical biomaterials, and in particular relates to an articular cartilage graft and a preparation method thereof. Background technique [0002] Articular cartilage is hyaline cartilage composed of chondrocytes, cartilage fibers, and cartilage matrix. Cartilage tissue has the characteristics of heterogeneous layer distribution, which is divided into superficial layer (10-20%), middle layer (40-60%) and deep layer (20-30%) from top to bottom. In recent years, studies have found that each region of cartilage has a different structure, different protein components, and performs different functions. The gene expression and growth rate of chondrocytes obtained from different regions are different. The density of chondrocytes in the superficial, middle and deep layers decreases in turn, among which the chondrocytes in the superficial layer are closely arranged in the horizontal direction and ...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/38A61L27/24A61L27/22A61L27/20A61L27/54A61F2/30
Inventor 丛丽媛刘影
Owner 西安博鸿生物技术有限公司
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