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Assembling type cell-derived extracellular matrix membrane compound bone repairing material as well as preparation method and application thereof

An acellular matrix and extracellular matrix technology, which is applied in the field of assembled cell-derived extracellular matrix membrane composite bone repair material and its preparation, can solve the problem of difficult standardized production of classical bone tissue engineering products, no systematic report, stem cell tumorigenicity and other problems, to achieve the effects of good osteoinductive ability, enhanced bone regeneration ability, and rapid assembly

Active Publication Date: 2018-07-24
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, due to the complexity and dynamics of ECM, there is no systematic report on the use of CDM to construct ECM composites with controllable ECM components, but it will undoubtedly become a hot research direction for tissue engineering materials.
[0008] In addition, the traditional bone tissue engineering strategy of using BMSCs as seed cells combined with biomaterials and growth factors has certain limitations.
The classic bone tissue construction strategy takes a long time from the acquisition, expansion, planting, and induced differentiation of primary cells, requires a lot of manpower and material resources, and involves biological pollution and tumorigenicity of stem cells, making classic bone tissue engineering products Difficult to standardize production and widely used clinically

Method used

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  • Assembling type cell-derived extracellular matrix membrane compound bone repairing material as well as preparation method and application thereof
  • Assembling type cell-derived extracellular matrix membrane compound bone repairing material as well as preparation method and application thereof
  • Assembling type cell-derived extracellular matrix membrane compound bone repairing material as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: Preparation of a kind of rat osteoinductive acellular matrix membrane wrapped collagen composite bone repair material (OiECM-COL composite bone repair material), its specific preparation process is as follows figure 1 As shown, the detailed steps are as follows:

[0049] (1) Preparation of osteoinductive decellularized matrix membrane (OiECM membrane):

[0050] a. Rat bone marrow mesenchymal stem cells (rat BMSCs, Cat. No. RAWMX-01001, Cyagen Bioscience) were planted in a petri dish, and osteogenic induction medium was added (medium formula: 90% DMEM, 10% calf serum, Penicillin 100U / mL, streptomycin 100μg / mL, L-ascorbic acid 50ug / mL, 10mmol / L β-glycerol phosphate, 100nmol / L dexamethasone, change the solution every three days, at a temperature of 37°C, CO 2 Continuous culture in an incubator with a level of 5% and a pH value of 7.3 for 21 days until the extracellular matrix is ​​secreted and covered the bottom of the dish;

[0051] b. Discard the culture ...

example 2

[0061] Example 2: Acellular Matrix Membrane Encapsulation of Adipose Stem Cells Preparation of composite bone repair materials, the three-dimensional porous collagen scaffold material is selected from DePuy company Its specific preparation process is as follows:

[0062] (1) Preparation of adipose-derived stem cell acellular matrix membrane (adECM membrane):

[0063] a. Rat adipose-derived stem cells (Wistar rat adipose-derived mesenchymal stem cells, Cat. No. RAWMD-01001, Cyagen Bioscience) were planted in a petri dish, and osteogenic induction medium (90% DMEM, calf 10% calf serum, Penicillin 100U / mL, streptomycin 100μg / mL, L-ascorbic acid 50ug / mL, 10mmol / Lβ-glycerol / L phosphate, 100nmol / L dexamethasone), change the medium once every three days, at a temperature of 37°C, CO 2 Continuous culture in an incubator with a level of 5% and a pH value of 7.3 for 21 days until the extracellular matrix is ​​secreted and covered the bottom of the dish;

[0064] b. Remove the cultu...

example 3

[0067] Example 3: Preparation of fibroblast acellular matrix membrane-wrapped chitosan composite bone repair material, the specific steps are as follows:

[0068] (1) Preparation of fibroblast decellularized matrix membrane (fbECM membrane):

[0069] a. Human fibroblasts (human fibroblasts, product number HXXFB-00001, Cyagen Bioscience) were planted in a petri dish, and conditioned medium (90% DMEM, 10% calf serum, penicillin 100U / mL, streptomycin 100μg / mL L-ascorbic acid, 20-100ug / mL L-ascorbic acid), change the solution every three days, at a temperature of 37°C, CO 2 Continuous culture in an incubator with a level of 5% and a pH value of 7.3 for 14 days until the extracellular matrix is ​​secreted and covered the bottom of the dish;

[0070] b. Remove the culture medium from the cell culture dish in step a and gently rinse it with PBS solution for 3 times, add 0.1% SDS to the culture dish and treat it at 37°C for 60 minutes;

[0071] c. Rinse the cell culture dish with PB...

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Abstract

The invention provides an assembling type cell-derived extracellular matrix membrane (ECM) compound bone repairing material as well as a preparation method and application thereof. The bone repairingmaterial is prepared by the following steps: completely covering a three-dimensional degradable biological scaffold with a sheet-shaped acellular matrix membrane under sterile conditions and then freezing the three-dimensional degradable biological scaffold for 24 to 48h under the condition of -20 to -80 DEG C; then carrying out freeze drying treatment to obtain the ECM membrane compound bone repairing material, wherein the sheet-shaped acellular matrix membrane is a cell-derived extracellular matrix which is generated by in-vitro culture of cells, is subjected to decellularization treatment and has certain bone induction activity, and the three-dimensional degradable biological scaffold is a biological scaffold which is prepared from natural polymers, has a three-dimensional porous microscopic structure and can be completely degraded and absorbed in human bodies. The assembling type cell-derived extracellular matrix membrane compound bone repairing material has a good bone induction capability; furthermore, the bone regeneration capability of the compound material is strengthened; a construction method is simple and feasible and the material can be stored for a long time and has awide clinical application potential.

Description

technical field [0001] The invention relates to the field of bone tissue engineering biomaterials, in particular to an assembled cell-derived extracellular matrix membrane composite bone repair material and a preparation method thereof. Background technique [0002] So far, the repair of large bone defects caused by trauma, infection and tumor resection has been a long-standing problem in clinical medicine at home and abroad. Autologous bone sources are limited, and patients may need to bear additional pain, while other bone repair materials, such as allogeneic bone or xenograft bone, are prone to potential infection and immune rejection. Therefore, the research and development of bone repair materials (or bone graft substitute materials) that can replace autogenous bone is a recent research hotspot. [0003] At present, the commonly used bone graft repair materials in clinical practice mainly include polymers, bioceramics, and magnesium-based biodegradable biomaterials and...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/24A61L27/20A61L27/50A61L27/56A61L27/58
CPCA61L27/20A61L27/24A61L27/3633A61L27/3687A61L27/50A61L27/56A61L27/58A61L2430/02C08L5/08
Inventor 刘绍恺汪振星孙家明牟珊陈雳风周牧冉曾宇阳李嘉伦周楚超罗超黎媛侯金飞方慧敏
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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