Preparation method and application of cell-stent composite material for cartilage injury repairing

A composite material and cartilage damage technology, applied in the field of bone tissue engineering and regenerative medicine, can solve the problem of not finding materials and methods for repairing cartilage damage

Inactive Publication Date: 2017-11-24
中国人民解放军联勤保障部队第九二〇医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are still no materials and methods for cartilage damage repair that are good in terms of cells, scaffolds, and tissue biocompatibility.

Method used

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  • Preparation method and application of cell-stent composite material for cartilage injury repairing
  • Preparation method and application of cell-stent composite material for cartilage injury repairing
  • Preparation method and application of cell-stent composite material for cartilage injury repairing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Fish oocyte extract induces reprogramming of rabbit skin fibroblasts into induced pluripotent stem cells.

[0040] In this example, the fibroblasts were derived from the skin tissue on the back of rabbits, and the technical route for the research of fish oocyte extracts to induce the reprogramming of rabbit skin fibroblasts into induced pluripotent stem cells is shown in the attached figure 1 shown, including the following steps.

[0041] (1) Preparation of fish oocyte extract Select the local crucian carp produced in Kunming when the oocyte yolk is full, soak it in 75% alcohol for 15-20 minutes, take out the fish eggs under sterile conditions, and put them in pre-cooling Put the mortar on ice for grinding. After fully grinding, add pre-cooled saline at a volume ratio of about 1:1, centrifuge at 1000rpm at 4°C for 10min, and then centrifuge at 3000rpm at 4°C. 10min, take the supernatant, use a 0.22 micron pore filter to filter, use the Coomassie brilliant blue method t...

Embodiment 2

[0048] Preparation of KGM / HA bioscaffolds.

[0049]The preparation materials of the KGM / HA bioscaffold in this example include konjac glucomannan and sodium hyaluronate. The preparation method is as follows: disperse 0.06g HA powder in 40 ml water, stir magnetically, and add 0.08ml NH 3 ·H 2 O adjust the pH value, add 1.50g of KGM according to the design of the orthogonal test table, and continue magnetic stirring at the same time until the stirring stops automatically and seals; it is matured in a water bath at 70°C, and dealkalized under the same conditions; then it is placed in a low-temperature freezer. , frozen at -20°C for 24 hours, then taken out, and finally dried in a freeze dryer to obtain the composite porous scaffold KGM / HA.

[0050] Using optical microscopic analysis and quantitative metallographic analysis, it was found that the prepared KGM / HA scaffold was suitable for the growth of stem cells. The pore size of 100 μm to 250 μm accounted for 80% to 90% of the t...

Embodiment 3

[0052] Preparation of reprogrammed cells and KGM / HA scaffold composites.

[0053] Co-cultivate the reprogrammed cells prepared in Example 1 and the KGM / HA scaffold prepared in Example 2 to prepare the reprogrammed cell-KGM / HA scaffold composite material, the method and steps are as follows: (1) Implement the The aseptically treated KGM / HA scaffold prepared in Example 2 was placed in a six-well plate for cell culture; (2) Suspended in the DMEM / F12 medium containing 10% fetal bovine serum prepared in Example 1 of reprogrammed cells and adjust their concentration to 1 x 10 per ml 6 (3) inoculate the reprogrammed cells with adjusted concentration on the sterile KGM / HA composite material in the six-well plate in the step (1), inoculate 5ml in each well; (4) inoculate the Place the six-well culture plate seeded with reprogrammed cells in step (3) at 37°C, 5% CO 2 1. Cultivate under 90% humidity conditions, replace the culture medium every two days, and cultivate for 10 days; (5) A...

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Abstract

The invention provides a preparation method and application of a cell-stent composite material for cartilage injury repairing, and belongs to the fields of bone tissue engineering and regenerative medicines. Reprogrammed cells with multi-directional differentiation potential and relatively good security are adopted as seed cells for preparing the cell-stent composite material, a biological material which is prepared from konjac glucomannan and sodium hyaluronate through mixing is adopted as a stent, and the cell-stent composite material is prepared from the cells and the stent by using a co-culture method. The cell-stent composite material is relatively simple in preparation method, is capable of effectively repairing cartilage injury, and has wide application prospects in the fields of bone tissue engineering and regenerative medicines, particularly in the field of joint cartilage repairing.

Description

technical field [0001] The invention relates to a preparation method and application of a cell-scaffold composite material for repairing cartilage damage, and belongs to the fields of bone tissue engineering and regenerative medicine. Background technique [0002] Cartilage tissue is a tissue rich in proteoglycans, collagen, and hyaluronic acid, but lacks blood supply and chondrocytes. Therefore, once cartilage tissue is damaged, it is difficult to repair or heal naturally even a small cartilage injury. At present, the commonly used repair methods for articular cartilage injury in clinic mainly include cartilage and subchondral bone removal, microfracture and subchondral bone drilling, autologous osteochondral transplantation and allogeneic osteochondral transplantation, cartilage or periosteum transplantation, etc. (Liu imitation, et al. Chinese Journal of Traumatology and Orthopedics, 2014,16(6):537-539). Among these methods, the autologous osteochondral transplantation m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/26A61L27/58
CPCA61L27/26A61L27/3804A61L27/3852A61L27/58C08L5/08C08L5/02
Inventor 潘兴华朱向情陈强陈庆华
Owner 中国人民解放军联勤保障部队第九二〇医院
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