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Mineralized collagen bionic bone repair material modified by hyaluronan oligosaccharide and preparation method of mineralized collagen bionic bone repair material

A hyaluronic acid oligosaccharide and collagen biomimetic bone technology, which is applied in the fields of analysis materials, tissue regeneration, instruments, etc., can solve the problems of multiple operations and unrealized bone microstructure biomimetic, achieve low cost and improve biocompatibility , the effect of simple method

Active Publication Date: 2017-06-20
SHANDONG UNIV
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  • Claims
  • Application Information

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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

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  • Mineralized collagen bionic bone repair material modified by hyaluronan oligosaccharide and preparation method of mineralized collagen bionic bone repair material
  • Mineralized collagen bionic bone repair material modified by hyaluronan oligosaccharide and preparation method of mineralized collagen bionic bone repair material
  • Mineralized collagen bionic bone repair material modified by hyaluronan oligosaccharide and preparation method of mineralized collagen bionic bone repair material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Preparation of glycosylated collagen (Col / HA, HA is hyaluronic acid raw material before enzymatic hydrolysis):

[0065] 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;

[0066] 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;

[0067] 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

[0069] Preparation of glycosylated collagen (Col / oHAs, oHAs are oligosaccharides after enzymatic hydrolysis):

[0070] 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.

[0071] 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%.

[0072] 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.

[0073] For the preparation steps of hyaluronic acid oligosaccharides, see "Optimum Conditions for Hyaluronic Acid Hydrolysis Catalyzed by Hyaluronid...

Embodiment 3

[0075] Preparation of Glycosylated Collagen Mineralization Composite (Col / oHAs / HAP):

[0076] 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 mineralized collagen bionic bone repair material modified by hyaluronan oligosaccharide and a preparation method of the mineralized collagen bionic bone repair material. The mineralized collagen bionic bone repair material modified by hyaluronan oligosaccharide has the following structure: collagen in a collagen-hydroxyapatite composite material is connected with hyaluronan oligosaccharide through C / N bonds, thus obtaining the mineralized collagen composite material with galactosylated modification, wherein the molecular weight of the hyaluronan oligosaccharide is 776-5000 Da. According to the mineralized collagen bionic bone repair material and the preparation method, Schiff base reaction is adopted for carrying out hyaluronan oligosaccharide modification on the collagen for the first time, the glycosylated collagen realizing covalent binding is obtained, meanwhile, the glycosylated collagen is taken as a mineralization template to be applied to the design of the bone scaffold for the first time, the functions facilitating cell migration, proliferation and differentiation and promoting wound healing of low-molecular-weight HA are exerted, and a novel material basis and research strategy are provided for the in-vitro construction of the vascularized stent.

Description

technical field [0001] 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 [0002] 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 bone, xenograft bone, artificial bone, etc., but there are certain problems and cannot completely solve the huge clinical demand for bone materials. Although there are limited sources and secondary trauma, autogenous bone is still the bone implant material with the best clinical effect, and has...

Claims

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Application Information

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IPC IPC(8): A61L27/46A61L27/50A61L27/56D01D5/00
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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