Acellular bone matrix hydrogel with reserved natural hydroxyapatite and preparation method of acellular bone matrix hydrogel

A kind of acellular bone matrix, hydroxyapatite technology, applied in medical science, prosthesis, tissue regeneration and other directions, can solve the problems of easy agglomeration, uneven dispersion, limited addition of hydroxyapatite, etc., to achieve good storage stability The effect of stability, uniform dispersion and good injectability

Active Publication Date: 2020-11-10
北京欧亚铂瑞科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

If you want to obtain organic-inorganic composite ECM hydrogels for bone regeneration repair, usually only by adding hydroxyapatite to ECM hydrogels, mineralizing ECM gels with simulated body fluids, or gels with hydroxyapatite Co-precipitation and other methods introduce hydroxyapatite components to achieve, but these preparation methods often have shortcomings such as easy agglomeration of hydroxyapatite, uneven dispersion, and limited addition, which limits its clinical use

Method used

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  • Acellular bone matrix hydrogel with reserved natural hydroxyapatite and preparation method of acellular bone matrix hydrogel
  • Acellular bone matrix hydrogel with reserved natural hydroxyapatite and preparation method of acellular bone matrix hydrogel
  • Acellular bone matrix hydrogel with reserved natural hydroxyapatite and preparation method of acellular bone matrix hydrogel

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Embodiment 1

[0060] A method for preparing a decellularized bone matrix hydrogel retaining natural hydroxyapatite, comprising the following steps:

[0061] a. Choose fresh sheep cortex bone, rinse repeatedly with high-speed water to remove bone marrow, freeze-dry for 24 hours, crush, and sieve to obtain bone powder with a particle size of less than 0.1 mm. Soak the bone powder in 75% ethanol for 1 hour to sterilize, and wash with PBS buffer. 3 hours;

[0062] b. Soak the bone powder obtained in step a with a mixture of chloroform and methanol at a volume ratio of 1:1 for 2 h, the ratio of bone powder to chloroform / methanol mixture solution is 1:25 (w / v), and wash with PBS buffer for 3 hours Then soak the bone powder in pH 7.4 10 mM Tris-HCl buffer solution for 12 hours, the mass volume ratio of bone powder and Tris-HCl buffer solution is 1:25 (w / v) to obtain defatted bone powder;

[0063] c. Soak the defatted bone powder prepared in step b in 10 mM Tris-HCl buffer solution containing 0.8%...

Embodiment 2

[0069] A method for preparing a decellularized bone matrix hydrogel retaining natural hydroxyapatite, comprising the following steps:

[0070] a. Choose fresh bovine cortical bone, rinse repeatedly with high-speed water to remove bone marrow, freeze-dry for 24 h, crush, and sieve to obtain bone powder with a particle size of less than 1 mm. Soak the bone powder in 75% ethanol for 0.5 h to sterilize, and buffer with PBS. Wash with liquid for 6 h;

[0071] b. Soak the bone meal obtained in step a for 3 hours in a mixture of chloroform and methanol with a volume ratio of 1:0.5, the ratio of bone meal to chloroform / methanol mixture solution is 1:25 (w / v), and wash with PBS buffer for 3 hours Then soak the bone powder in pH 7.4 10 mM Tris-HCl buffer solution for 14 hours, the mass volume ratio of bone powder and Tris-HCl buffer solution is 1:25 (w / v) to obtain defatted bone powder;

[0072] c. Soak the defatted bone powder prepared in step b in pH 7.4 10 mM Tris-HCl buffer contain...

Embodiment 3

[0079] A method for preparing a decellularized bone matrix hydrogel retaining natural hydroxyapatite, comprising the following steps:

[0080] a. Select fresh sheep cancellous bone, rinse repeatedly with high-speed water to remove bone marrow, freeze-dry for 24 hours, pulverize, and sieve to obtain bone powder with a particle size of less than 0.1 mm. Soak the bone powder in 75% ethanol for 1.5 h to sterilize, and wash with PBS Wash with buffer for 1 h;

[0081] b. Soak the bone meal obtained in step a in the mixed solution of chloroform and methanol with a volume ratio of 1:2 for 1 hour, the ratio of bone meal and chloroform / methanol mixed solution is 1:25 (w / v), and wash with PBS buffer for 3 times ; Soak the bone powder in pH 7.4 10 mM Tris-HCl buffer solution for 10 h, the mass volume ratio of bone powder and Tris-HCl buffer solution is 1:25 (w / v), to obtain defatted bone powder;

[0082] c. Soak the defatted bone powder prepared in step b in pH 7.4 10 mM Tris-HCl buffer ...

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Abstract

The invention discloses acellular bone matrix hydrogel with reserved natural hydroxyapatite and a preparation method of the acellular bone matrix hydrogel, and belongs to the field of bone regeneration and repair materials in tissue engineering. According to the organic-inorganic composite hydrogel, cortical bones or cancellous bones from the same source or different sources are used as raw materials, natural bone minerals of different degrees are reserved through degreasing, decellularization and decalcification treatment, then enzymolysis digestion and purification treatment are conducted, the pH and salt concentration of a solution are adjusted to physiological conditions, and then a pre-gel solution is obtained; and when in use, the temperature is raised to 37 + / -1 DEG C, and the hydrogel is formed through self-assembly. According to the bone tissue decellularized extracellular matrix hydrogel, natural hydroxyapatite components in bone matrixes are effectively reserved, hydroxyapatite and collagenous fibers are well combined and uniformly dispersed, and the bone tissue decellularized extracellular matrix hydrogel can be used as temperature-sensitive injectable hydrogel to be applied to bone tissue engineering and has bright application prospects.

Description

technical field [0001] The application relates to the field of bone regeneration and repair materials in tissue engineering, in particular to a decellularized bone matrix hydrogel retaining natural hydroxyapatite and a preparation method thereof. Background technique [0002] Currently, the clinical treatment of critical bone defects caused by trauma, infection, tumor, or congenital disease still tends to be based on autologous or allogeneic bone grafts. However, issues such as severe donor site morbidity, high risk of infection, and transportation hinder their development as sustainable bone repair materials. [0003] Tissue engineering technology based on biodegradable porous scaffolds is considered to be a feasible and effective alternative strategy for bone regeneration repair. Nowadays, the materials that have been widely used in bone tissue engineering scaffolds are mostly synthetic degradable aliphatic polyesters, such as polylactic acid, poly(lactic acid-glycolic ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/12A61L27/20A61L27/24A61L27/36A61L27/50A61L27/52A61L27/54A61L27/58
CPCA61L27/12A61L27/20A61L27/24A61L27/3633A61L27/365A61L27/3687A61L27/50A61L27/52A61L27/54A61L27/58A61L2300/412A61L2430/02A61L2430/40C08L5/00
Inventor 蔡晴左大为易蜜杨小平
Owner 北京欧亚铂瑞科技有限公司
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