Nanometer hydroxyapatite/chitosan porous composite scaffold material as well as bionic dialysis mineralization preparation method and application thereof

A technology of nano-hydroxyapatite and composite scaffolds, applied in tissue regeneration, medical science, prostheses, etc., can solve the problems of poor host bonding ability, low matching degree of degradation absorption rate and growth rate, low mechanical strength and toughness, etc. , to achieve controllable degradability, excellent bone conduction, and full-featured effects

Inactive Publication Date: 2018-09-04
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chitosan is often used clinically to prepare bone tissue engineering scaffold materials. A large number of experimental studies have also confirmed that CS scaffold composite cells or scaffold materials constructed with cytokines have good osteogenic ability in vivo, but chitosan There are still shortcomings that cannot be ignored when used for bone defect repair: such as (1) low mechanical strength and toughness; (2) the degradation and absorption rate of chitosan in the body is relatively low in matching with the growth rate of regenerated tissue; (3) and The host has poor bonding ability, etc.

Method used

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  • Nanometer hydroxyapatite/chitosan porous composite scaffold material as well as bionic dialysis mineralization preparation method and application thereof
  • Nanometer hydroxyapatite/chitosan porous composite scaffold material as well as bionic dialysis mineralization preparation method and application thereof
  • Nanometer hydroxyapatite/chitosan porous composite scaffold material as well as bionic dialysis mineralization preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] A preparation method of n-HAP porous composite support material, comprising the following steps:

[0059] (1) the chitosan powder that takes 2.5g is dissolved in the acetic acid solution of 50mL 3% (v / v), and stirs 24h at a constant speed, after chitosan dissolves completely, the final concentration that obtains chitosan is 5% ( Wt). Add the chitosan solution into the 96-well plate with a 2.5mL needle-free syringe, add 0.5mL to each well, let it stand for a while, and put the orifice plate into a 70°C refrigerator after there are no air bubbles in each well Freezing and icing, and finally freeze-drying at low temperature to obtain the porous chitosan scaffold in the early stage. Soak the dried stent together with the orifice plate in a 5% NaOH solution for 24 hours, soak in distilled water and wash the stent repeatedly until the pH is about 7-8, then freeze-dry at low temperature again to obtain the final solution required for this experiment. chitosan porous scaffold...

Embodiment 2

[0065] A preparation method of n-HAP porous composite support material, comprising the following steps:

[0066] (1) the chitosan powder that takes 2.5g is dissolved in the acetic acid solution of 50mL 3% (v / v), and stirs 24h at a constant speed, after chitosan dissolves completely, the final concentration that obtains chitosan is 5% ( Wt). Add the chitosan solution into the 96-well plate with a 2.5mL needle-free syringe, add 0.5mL to each well, let it stand for a while, and put the orifice plate into a 70°C refrigerator after there are no air bubbles in each well Freeze and freeze in medium, and finally freeze-dry at low temperature to obtain the CS scaffold in the early stage. Soak the dried CS scaffold together with the orifice plate in NaOH solution with a mass fraction of 5% for 24 hours, soak in distilled water and wash the scaffold repeatedly until the pH=7-8, then freeze-dry at low temperature again to obtain the final Desired to obtain a mineralized organic template...

Embodiment 3

[0073] A preparation method of n-HAP porous composite support material, comprising the following steps:

[0074] (1) Prepare the n-HAP / CS composite scaffold material according to the material preparation method in the above-mentioned Example 2, and mark the material as dialysis 24h, dialysis 48h and dialysis 72h according to the different dialysis time.

[0075] (2) Select the mineralized scaffold material and pure CS scaffold as the research object of the cell culture experiment, sterilize the scaffold material with cobalt 60 (radiation intensity is 15k gy), soak it with 75% alcohol on the ultra-clean bench After 30 minutes, rinse with PBS three times for later use. Take the third-generation MC3T3-E1 cells. After the density reaches 90%, digest with 0.25% trypsin for 3 minutes, stop the digestion with 2 times the volume of complete medium, and centrifuge at 1000r / min. Cells were collected for 5 minutes and resuspended in complete medium. Dilute the above treated cell suspens...

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Abstract

The invention belongs to the technical field of bone tissue engineering, and discloses a nanometer hydroxyapatite/chitosan porous composite scaffold material as well as a bionic dialysis mineralization preparation method and application thereof. The method comprises the following steps that (1) chitosan is dissolved in an acetic acid solution to obtain a chitosan solution; the chitosan solution isput into a container for freezing; freeze drying is performed to obtain an early-period chitosan porous scaffold; (2) the early-period chitosan porous scaffold is soaked in alkali liquid; water washing is performed until the pH is 7 to 8; freeze drying is performed to obtain a chitosan porous scaffold; (3) the chitosan porous scaffold is soaked in an alkaline phosphatase solution; then, the chitosan porous scaffold and the alkaline phosphatase solution are charged into a dialysis bag; the dialysis bag is put into a mixed solution of calcium glycerophosphate and CaCl2; constant-temperature constant-speed stirring is performed for mineralization; taking out, washing and freeze drying are performed to obtain the nanometer hydroxyapatite/chitosan porous composite scaffold material. The nanometer hydroxyapatite/chitosan porous composite scaffold material can be applied to the field of bone tissue engineering, particularly the bone defect repair.

Description

technical field [0001] The invention belongs to the technical field of bone tissue engineering, and in particular relates to a nano-hydroxyapatite / chitosan (n-HAP / CS) porous composite scaffold material and a bionic dialysis mineralization preparation method and application thereof. Background technique [0002] Bone is a hard organ of the human body with high hardness and certain flexibility. It is composed of periosteum, bone tissue and bone marrow, and has abundant blood vessels and nerve distribution. Bone tissue is characterized by the deposition and orderly arrangement of a large number of inorganic calcium salts in the extracellular matrix, which endows the bone with a strong organizational structure foundation. Bone is the support body of the whole human body. It has the functions of cooperating with body movements and protecting internal organs. It is also the main supply bank of calcium and phosphorus ions required to maintain the metabolism of the human body. Calc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/56A61L27/58A61L27/50A61L27/46
CPCA61L27/46A61L27/50A61L27/56A61L27/58A61L2430/02C08L5/08
Inventor 李立华李娜张舒昀周长忍
Owner JINAN UNIVERSITY
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