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Hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material

一种透明质酸寡糖、胶原仿生骨的技术，应用在生物医用材料领域，能够解决操作过程多、未实现骨微结构仿生等问题，达到成本低廉、改善生物相容性、方法简单的效果

Active Publication Date: 2020-01-03
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent document CN101590293A (application number 200910149959.0) discloses a preparation method of HA (this refers to hydroxyapatite) / collagen / chitosan interpenetrating polymer network scaffold, which uses PVP as a template to prepare hydroxyapatite The stone sol is dispersed in the collagen and chitosan blend, and after two-step cross-linking, the composite scaffold is obtained by post-treatment such as decompression and degassing. Although the inorganic particles in the scaffold are well dispersed in the collagen / chitosan matrix, the On the whole, there are many operations, and the natural bone uses collagen as a template to realize the nucleation and mineralization of calcium and phosphorus salts through the regulation of non-collagen protein or the interaction between collagen and mineral phase, and the formation is based on the self-assembly process of collagen The hierarchical structure of bone, which is formed step by step from microscopic to macroscopic, is very important to the performance and function of bone. However, in this literature, only the separately prepared hydroxyapatite is cross-linked and bonded to the polymer matrix. However, bionic bone microstructure has not been realized

Method used

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  • Hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material
  • Hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material
  • Hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Preparation of glycosylated collagen (Col / HA, HA is hyaluronic acid raw material before enzymatic hydrolysis):

[0066] Preparation of the reaction system: Weigh 80 mg of collagen, 20 mg of HA (5K), recrystallized NaBH 3 CN 30mg in a 25mL glass bottle, add the prepared reaction system (i.e. HFP and 0.1M NaHCO 3 Mixed medium, volume ratio 3:2) 5mL, under dark conditions, 37°C, magnetic stirring for 24h;

[0067] Cleaning of reaction samples: After the reaction, transfer the reaction solution to a small beaker, add 5% acetic acid for 8-fold dilution, and put it into six 30K ultrafiltration tubes, 4000g / min, 25min / time, ultrafiltration and centrifugal replacement 6 times, and then collect samples for vacuum freeze-drying;

[0068] Determination of the sugar loading of the reaction sample: the sugar content of the dried Col / HA was measured by the carbazole-ethanol method, and the sugar loading was determined to be 5.266%.

Embodiment 2

[0070] Preparation of glycosylated collagen (Col / oHAs, oHAs are oligosaccharides after enzymatic hydrolysis):

[0071] The preparation method as described in Example 1, the difference is that collagen 80mg, oHAs (776.5-2293.4) ​​20mg, recrystallized NaBH 3 CN 30mg, ultrafiltration and centrifugal replacement 4 times.

[0072] The sugar content of the dried Col / HA was measured by the carbazole-ethanol method, and the sugar loading was determined to be 2.101%.

[0073] oHAs is the glycosylation modification of collagen with hyaluronic acid oligosaccharides. Hyaluronic acid oligosaccharides are tetrasaccharides, hexasaccharides, octasaccharides, decasaccharides, and dodecasaccharides obtained after enzymatic hydrolysis, separation and purification of 5k sodium hyaluronate And its mixture, the molecular weight range is 776-2293Da.

[0074] For the preparation steps of hyaluronic acid oligosaccharides, please refer to "Optimum conditions for hyaluronic acid hydrolysis reaction ca...

Embodiment 3

[0076] Preparation of Glycosylated Collagen Mineralization Composite (Col / oHAs / HAP):

[0077] Weigh 60 mg of the dried glycosylated collagen (Col / oHAs) prepared in Example 2 into 100 mL of HCl (concentration 0.01M) solution, and dissolve with magnetic stirring to obtain the Col / HA solution, and slowly add CaCl dropwise while stirring 2 (0.1M) solution 14mL, after mixing, let it stand for 10min, continue to stir and slowly add NaH 2 PO 4 (0.1M) solution 8.4mL (Ca:P=1.66), then slowly add 0.1M NaOH dropwise during stirring to adjust the pH of the system. When the pH is close to 6, the solution begins to appear turbid, and when the pH is about 6.2, the fluctuation is large. When the reaction is still in progress, continue to add NaOH dropwise until the pH of the system is 7, then continue to stir for 2 hours and keep the pH of the mineralization system unchanged; Centrifuge at 8000r / min for 3 times (8min / time), freeze-dry the sample at last, and grind to obtain hyaluronic acid-...

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Abstract

The invention relates to a hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material and a preparation method thereof. The hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material has the following structure that collagen in a collagen-hydroxyapatite composite material is connected with hyaluronic acid oligosaccharide through aC-N bond, a glycosylation modified mineralization collagen composite material is obtained, and the molecular weight of the hyaluronic acid oligosaccharide is 776-5000Da. According to the hyaluronic acid oligosaccharide modified mineralization collagen bionic bone repair material disclosed by the invention, the collagen is subjected to hyaluronic acid oligosaccharide modification through a Schiff's base reaction for the first time, glycosylation collagen in covalent bonding can be obtained, the idea that the glycosylation collagen is used as a mineralization formwork to be used for bone stentdesign is put forward for the first time, and besides, the functions that low-molecular-weight HA facilitates cell migration, proliferation, disintegration and wound healing promotion are achieved, anew material base and a research strategy are provided for in vitro construction of a vascularization stent.

Description

[0001] This application is a divisional application of the following application: application date March 28, 2017, application number 2017101926909, invention name: a mineralized collagen biomimetic bone repair material modified with hyaluronic acid oligosaccharides and its preparation method technical field [0002] The invention relates to a mineralized collagen bionic bone repair material modified by hyaluronic acid oligosaccharides and a preparation method thereof, belonging to the technical field of biomedical materials. Background technique [0003] Bone defects caused by trauma, tumors, infections and pathological factors are very common in clinical practice and have become one of the problems that plague the healthy life of human beings. Bone transplantation has become the graft with the largest demand after blood transfusion, and there is an increasing trend year by year. At present, bone repair materials for clinical application include autologous bone, allogeneic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/46A61L27/50A61L27/56D01D5/00C12Q1/02
CPCA61L27/46A61L27/50A61L27/56A61L2430/02D01D5/003G01N33/5008C08L5/08
Inventor 陈宗刚张秀丽栗敏顾国锋郭忠武
Owner SHANDONG UNIV
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