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Method for preparing biomimetic artificial bone materials for biodegradable tissue engineering

A tissue engineering and biodegradation technology, applied in medical science, prosthesis, etc., can solve problems such as tissue damage, reduce workload, and shorten the preparation technology cycle

Active Publication Date: 2013-03-06
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because glutaraldehyde is highly toxic to cells, and its degradation products will also cause certain damage to body tissues after the bone repair material cross-linked with it is implanted in the body.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] At 100°C, Ca(NO 3 ) 2 solution with (NH 4 ) 3 PO 4 Solution reaction, the pH value of the reaction system is 0.1M NH 3 ·H 2 O was adjusted to keep the pH at 9±0.1, and the stirring reaction was continued for 4 hours to prepare a needle-shaped nano-hydroxyapatite suspension. The reaction formula is as follows:

[0030] Ca(NO 3 ) 2 4H 2 O+(NH 4 ) 3 PO 4 ·3H 2 O→Ca 10 (PO 4 ) 6 (OH) 2

[0031] The human-like collagen was dissolved into a 4% solution with distilled water, and the nano-hydroxyapatite suspension prepared above was added dropwise to the human-like collagen solution, so that the mass ratio of the human-like collagen to the nano-hydroxyapatite 6:4, stirred gently for 6h, stood overnight, and adjusted the pH of the solution to 7.0 with 0.1M NaOH. Add genipin to cross-link for 1 day, and vacuum freeze-dry for 24 hours to prepare recombinant human-like collagen II (RHLCII) / nano-hydroxyapatite (nHA) powder. Dissolve chitosan with dilute acetic ac...

Embodiment 2

[0033]The human-like collagen was dissolved into a 3% solution with distilled water, and the nano-hydroxyapatite suspension prepared above was added dropwise into the human-like collagen solution, so that the mass ratio of the human-like collagen to the nano-hydroxyapatite was 6:4, stirred gently for 6h, stood overnight, and adjusted the pH of the solution to 7.0 with 0.1M NaOH. Add genipin to cross-link for 1 day, and vacuum freeze-dry for 24 hours to prepare recombinant human-like collagen II (RHLCII) / nano-hydroxyapatite (nHA) powder. Dissolve chitosan with acetic acid into a 0.01% solution; add RHLCII / nHA powder dropwise to the chitosan solution, adjust the pH of the solution to 7.0 with 0.1M NaOH, add collagen again until the pH is 6.4, and add the aforementioned chitosan Sugar solution, adjust the pH of the solution to 7.0 with NaOH. The above steps are repeated four times, the total amount of chitosan added is 2% of the mass of the mixed solution, the mixed solution is ...

Embodiment 3

[0035] The human-like collagen was dissolved into a 4% solution with distilled water, and the nano-hydroxyapatite suspension prepared above was added dropwise into the human-like collagen solution, so that the mass ratio of the human-like collagen to the nano-hydroxyapatite was 6:4, stirred gently for 6h, stood overnight, and adjusted the pH of the solution to 7.0 with 0.1M NaOH. Add genipin to cross-link for 1 day, and vacuum freeze-dry for 24 hours to prepare recombinant human-like collagen II (RHLCII) / nano-hydroxyapatite (nHA) powder. Dissolve chitosan with dilute formic acid into a 0.01% solution; add RHLCII / nHA powder dropwise to the chitosan solution, adjust the pH of the solution to 7.0 with 0.1M NaOH, add collagen again to pH 6.4, and add the aforementioned shell The glycan solution was adjusted to pH 7.0 with NaOH. The above steps are repeated three times, the total amount of chitosan added is 3% of the mass of the mixed solution, the mixed solution is fully disperse...

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Abstract

The invention discloses a method for preparing biomimetic artificial bone materials for biodegradable tissue engineering. The method comprises the following steps: adding a hydroxyapatite suspension to a human-like collagen solution; adding Genipin for crosslinking; performing vacuum freeze-drying to obtain human-like collagen / hydroxyapatite powder; adding the human-like collagen / hydroxyapatite powder to a chitosan solution; using an NaOH solution to adjust the pH of the solution to 7.0; repeatedly adding the human-like collagen solution and the chitosan solution; adding a Genipin solution for crosslinking; freeze-drying a mixed solution; inoculating the obtained material with BMSCs of which the cell density is 5*10 cells / cm; performing culture for 1 to 10 days; using an induction medium to culture BMSCs cells so as to induce the BMSCs cells to differentiate towards an osteogenesis direction; and forming bone repair composite materials with bone inducing activity. The artificial bone materials prepared by the method are excellent in bone inducing activity and immune compatibility, thereby thoroughly putting an end to inevitable viral hidden trouble of animal collagen scaffold materials and greatly improving use safety.

Description

technical field [0001] The invention relates to a method for preparing a bionic artificial bone material for biodegradable tissue engineering, belonging to the field of biomedical materials. Background technique [0002] Biocompatibility is one of the important indicators to measure whether bone repair materials are excellent. Due to the good biocompatibility of natural biomaterials, researchers have applied them more in bone repair materials or bone tissue engineering in recent years, especially animal collagen. However, animal collagen is extracted from animal tissues, which cannot eliminate the hidden dangers of viruses such as mad cow disease, swine plague, and bird flu. The extraction of human collagen (from human placenta, bone, etc.) cannot eliminate the risk of infection such as hepatitis and AIDS. Therefore, the application of animal-derived or human-derived collagen is restricted by the World Health Organization. Furthermore, both animal-derived collagen and coll...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/40A61L27/38A61L27/54A61L27/58A61L27/24A61L27/20A61L27/12
Inventor 范代娣朱晨辉马晓轩米钰马沛朱晓丽薛文娇惠俊峰骆艳娥高琛
Owner NORTHWEST UNIV
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