Preparation method and application of cell-stent composite material for bone defect treatment and repairing

A technology of composite materials and scaffold materials, applied in the field of bone tissue engineering and regenerative medicine, can solve the problems of not finding cells and scaffolds

Inactive Publication Date: 2018-08-28
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 is still no material with excellent cell and scaffold properties, cell and scaffold, and tissue biocompatibility.

Method used

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  • Preparation method and application of cell-stent composite material for bone defect treatment and repairing
  • Preparation method and application of cell-stent composite material for bone defect treatment and repairing
  • Preparation method and application of cell-stent composite material for bone defect treatment and repairing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Fish oocyte extract induces reprogramming of canine skin fibroblasts into induced pluripotent stem cells.

[0038] In this example, the fibroblasts were derived from the skin tissue of the dog's back, and the fish oocyte extract induced the reprogramming of the canine skin fibroblasts into induced pluripotent stem cells, including the following steps.

[0039] (1) Preparation of fish oocyte extract: The adult cell reprogramming inducer was fish oocyte extract, and the fish oocytes were collected from local crucian carp in Kunming. The preparation method of the fish oocyte extract is as follows: soak crucian carp in 75% alcohol for 15-20 minutes, take out the fish eggs under sterile conditions, put them into a pre-cooled mortar, put the mortar on ice for Grind, then add pre-cooled 0.9% normal saline at a volume ratio of about 1:1, centrifuge at 1000rpm for 10min at 4°C, take the supernatant, then centrifuge at 3000rpm for 10min at 4°C, take the supernatant, Use a 0.22 m...

Embodiment 2

[0046] Preparation of HAP / β-TCP / CS bioscaffold.

[0047] The preparation material of the HAP / β-TCP / CS bioscaffold in this embodiment includes three single material components of HAP, β-TCP and CS, and the preparation method includes the following steps.

[0048] (1) Preparation of hydroxyapatite whiskers: prepare calcium and phosphorus ion concentration products with diammonium hydrogen phosphate solution and calcium nitrate solution [Ca 2+ ]·[PO 3 4- ]=0.0167mol 2 ·L -2 Add urea and template agent sorbitol, adjust the pH value of the solution to 2~3 with concentrated nitric acid, then react at a constant temperature in a water bath at 83°C for 10 hours, then raise the temperature to 93°C and react at a constant temperature for 6 hours, and age for 12 Hours later, the filter cake of whiskers was obtained by suction filtration, and dried at 70°C.

[0049] (2) Preparation of hydroxyapatite powder: prepare 1.2mol / L and 0.72mol / L Ca(NO 3 ) 2 and (NH 4 ) 2 HPO 4 Each aqu...

Embodiment 3

[0055] Preparation of seeded cells and HAP / β-TCP / CS scaffold composites.

[0056] The seed cells prepared in Example 1 and the HAP / β-TCP / CS scaffold prepared in Example 2 were co-cultured under specific conditions to prepare the cell-stent composite material, the method and steps are: (1) Place the aseptically treated HAP / β-TCP / CS scaffold prepared in Example 2 in a six-well cell culture plate; (2) use DMEM / F12 containing 10% fetal bovine serum The seed cells prepared in Example 1 were suspended in the medium, and the cell concentration was adjusted to be 1×10 per milliliter. 6 (3) Inoculate the seed cells with adjusted concentration on the sterile HAP / β-TCP / CS material in the six-well plate in the step (1), inoculate 5ml of seed cells in each well; (4) Inoculate the Place the six-well culture plate inoculated with seeded cells in step (3) at 37°C, 5% CO 2 1. Cultivate under 90% humidity conditions, replace the culture medium every 2 days, and co-culture for 10 days; (5) Aft...

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Abstract

The invention provides a preparation method and application of a cell-stent composite material for bone defect treatment and repairing and belongs to the field of bone tissue engineering and regenerative medicine. Reprogramming cells which are induced by adopting a roe mother cell extract and has multi-directional differentiation potential and good safety are adopted for seed cells for preparing the cell-stent composite material, the composite material prepared by mixing three materials including hydroxyapatite, beta-tricalcium phosphate and chitosan is adopted for a stent, the composite material has the advantages of good biocompatibility, high compressive resistance, moderate degradation speed and the like, and the prepared cell-stent composite material can be used for effectively treating and repairing the bone defect. The cell-stent composite material has wide application prospect in the field of bone tissue engineering and regenerative medicine, in particular to bone defect treatment and repairing.

Description

technical field [0001] The invention discloses a preparation method and application of a cell-scaffold composite material for bone defect treatment and repair, and belongs to the field of bone tissue engineering and regenerative medicine. Background technique [0002] In recent years, there are millions of patients with bone defects caused by trauma, infection and tumor in our country every year. Usually, severe bone defects cannot heal and repair by themselves, and must be restored effectively with the help of bone repair materials. Traditional autologous bone transplantation not only causes additional physical trauma to patients, but also brings serious mental and economic burdens to patients. Although allogeneic bone can be used as a therapeutic material to replace autologous bone in clinical practice, it is difficult due to donor limitations. At the same time, it is difficult to become a long-term strategy for the treatment of bone defects due to the problems of allogra...

Claims

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Application Information

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IPC IPC(8): A61L27/58A61L27/56A61L27/50A61L27/12A61L27/20A61L27/38
CPCA61L27/12A61L27/20A61L27/3834A61L27/3847A61L27/3895A61L27/50A61L27/56A61L27/58A61L2430/02C08L5/08
Inventor 潘兴华朱向情陈庆华陈强
Owner 中国人民解放军联勤保障部队第九二〇医院
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