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Preparation method of novel tissue repair-promoting material and application thereof

A technology for tissue repair and seeding cells, which is applied in tissue regeneration, medical preparations with inactive ingredients, and medical preparations containing active ingredients, etc. It can solve the problems of bone defects that cannot heal and repair by themselves.

Active Publication Date: 2019-11-22
JINAN UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Usually, severe bone defects cannot heal and repair by themselves, and must be restored effectively with the help of bone repair materials

Method used

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  • Preparation method of novel tissue repair-promoting material and application thereof
  • Preparation method of novel tissue repair-promoting material and application thereof
  • Preparation method of novel tissue repair-promoting material and application thereof

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preparation example Construction

[0055] In a preferred embodiment, the present invention provides a method for preparing a composite scaffold for bone defect treatment and repair, which specifically includes the following steps.

[0056] Preparation of TGF-β3-chitosan-collagen microspheres:

[0057] (1) Prepare microspheres by emulsification method, water phase: 1-3w% chitosan (CS) acetic acid aqueous solution and collagen aqueous solution with a concentration of 1-4mg / mL, oil phase: 14mL liquid paraffin, 10mL petroleum ether , 0.72mL span 80;

[0058] (2) Take 1-4mL CS solution and 1mL collagen solution (CS solution: collagen solution (v / v) = 1-4:1), mix well and add to the oil phase drop by drop, magnetically stir for 1500r / min, after mixing, add 1.5mL of 25% glutaraldehyde at 35°C for cross-linking, then let stand overnight, centrifuge at 3500g to remove the supernatant;

[0059] (3) The precipitate was washed successively with petroleum ether for 3 times, methanol for 2 times, acetone for 1 time, isopr...

Embodiment 1

[0085] Preparation of TGF-β3-chitosan-collagen microspheres:

[0086] (1) Prepare microspheres by emulsification method, water phase: 2w% chitosan (Zhengzhou Kerui Fine Chemical Co., Ltd., molecular weight 3.6KD, deacetylation degree 50%) (CS) acetic acid aqueous solution and concentration of 4mg / mL Collagen (Dongguan Hanshuo Jianyuan Biotechnology Co., Ltd.) aqueous solution, oil phase: 14mL liquid paraffin, 10mL petroleum ether, 0.72mL span 80;

[0087] (2) Take 4mL CS solution and 1mL bone collagen solution (CS solution: bone collagen solution (v / v) = 4:1), mix well, add dropwise to the oil phase, stir magnetically at 1500r / min, mix After homogenization, add 1.5mL of 25% glutaraldehyde to carry out cross-linking at 35°C, then let stand overnight, and centrifuge at 3500g to remove the supernatant;

[0088] (3) Rinse the precipitate with petroleum ether (3), methanol (2), acetone (1), isopropanone (1), ethanol (1), distilled water (2) in turn to obtain chitosan / collagen micr...

Embodiment 2

[0092] Preparation of chitosan sponge containing human-like collagen-EGF fusion protein:

[0093] (1) The preparation method of the humanoid collagen-EGF fusion protein is the same as that described in the patent CN 104098701B (201410352444.1);

[0094] (2) the chitosan solution that concentration is 2w% obtains chitosan sponge by the method for freeze-drying, then fully soaks the chitosan sponge of freeze-drying in the human-like collagen-EGF in (1) at low temperature fusion protein solution (12 μg / mL), and finally freeze-dried in a freeze dryer to obtain a chitosan sponge containing human-like collagen-EGF fusion protein.

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Abstract

The invention provides a preparation method of a novel tissue repair-promoting material and an application thereof, particularly can be used for bone defect and repair treatment, which belong to the field of tissue engineering and regenerative medicine. A composite scaffold is prepared by mixing and freeze-drying TGF-beta3-chitosan-ossein protein microspheres and chitosan sponge containing bioactive human-like collagen protein. The composite scaffold has the characteristics of being good in biocompatibility, is capable of promoting proliferation and adhesion of cells around injured tissues andcollecting stem cells in vivo, and is suitable for cell growth and osteogenic differentiation and the like. Meanwhile, the periodontal ligament stem cells can be tightly combined with the scaffold material, so that the good growth of the cells in the scaffold material is facilitated, and the scaffold material has stronger osteogenic differentiation capacity and can be better applied to treatmentof bone defects. Wide application prospects are realized in the fields of bone tissue engineering and regenerative medicine, particularly in the aspects of bone defect treatment and repair and the like.

Description

technical field [0001] The invention belongs to the field of tissue engineering and regenerative medicine, and more specifically provides a novel tissue-promoting repair material, its preparation method and application, especially for bone defect and repair treatment. Background technique [0002] Bone defect refers to a disease in which the integrity of bone structure is destroyed due to trauma, infection, tumor, congenital disease, etc., and is a common clinical symptom. Usually, severe bone defects cannot heal and repair by themselves, and must be restored effectively with the help of bone repair materials. In recent years, the rapid development of bone tissue regeneration engineering technology provides a new way for the treatment and repair of bone defects. Current research shows that the common substitute materials for bone tissue defect and repair mainly include natural polymer materials, synthetic polymers, inorganic materials, nanomaterials, etc. Each of the above...

Claims

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Application Information

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IPC IPC(8): A61L27/20A61L27/22A61L27/24A61L27/38A61L27/54A61L27/56A61K9/16A61K38/18A61K38/19A61K38/39A61K47/36A61P19/08
CPCA61L27/20A61L27/225A61L27/24A61L27/227A61L27/3834A61L27/3847A61L27/56A61L27/54A61K9/1652A61K9/167A61K38/39A61K38/1841A61K38/19A61K47/36A61P19/08A61L2430/02A61L2300/252A61L2300/412C08L5/08C08L89/00
Inventor 项琪杨艳黄亚东李杨帆肖巧学
Owner JINAN UNIVERSITY
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