CN107376012B - Chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and preparation method thereof - Google Patents
Chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and preparation method thereof Download PDFInfo
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- CN107376012B CN107376012B CN201710433540.2A CN201710433540A CN107376012B CN 107376012 B CN107376012 B CN 107376012B CN 201710433540 A CN201710433540 A CN 201710433540A CN 107376012 B CN107376012 B CN 107376012B
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- chitosan
- tissue engineering
- dimensional tissue
- engineering scaffold
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- 229920001661 Chitosan Polymers 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 8
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000012046 mixed solvent Substances 0.000 claims abstract description 4
- 238000004108 freeze drying Methods 0.000 claims abstract description 3
- 238000000926 separation method Methods 0.000 claims abstract description 3
- 238000007710 freezing Methods 0.000 claims description 4
- 230000008014 freezing Effects 0.000 claims description 4
- 239000002121 nanofiber Substances 0.000 abstract description 9
- 239000002904 solvent Substances 0.000 abstract description 4
- 210000001519 tissue Anatomy 0.000 description 12
- 239000002253 acid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000005191 phase separation Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000024245 cell differentiation Effects 0.000 description 2
- 238000010041 electrostatic spinning Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229920002101 Chitin Polymers 0.000 description 1
- 229920001410 Microfiber Polymers 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000003658 microfiber Substances 0.000 description 1
- 239000002062 molecular scaffold Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dispersion Chemistry (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention relates to a chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and a preparation method thereof, which adopts a ternary mixed solvent of acetic acid, 1, 4-dioxane and water with a certain proportion to dissolve chitosan, and can obtain the chitosan micro-nano fibrous three-dimensional tissue engineering scaffold after sample separation, freeze forming and freeze drying. The coexistence microstructure of the micro-nano fibers endows the scaffold with good biocompatibility and mechanical strength. The preparation method has simple process and good formability, and compared with the chitosan bracket prepared by the traditional binary solvent, the microstructure of the chitosan bracket is in the form of micro-nano fibers.
Description
Technical Field
The invention relates to a chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and a preparation method thereof, belonging to the technical field of biomedical materials.
Background
The chitosan is natural alkaline polysaccharide obtained by amination of chitin, has good biocompatibility and has important application in the field of tissue engineering materials. The tissue engineering scaffold prepared from chitosan has good mechanical property, is beneficial to the proliferation and differentiation of cells, has the characteristics of antibiosis and antiphlogosis, and is widely regarded by researchers, Sundarajan V and the like firstly adopt a phase separation technology to prepare the chitosan scaffold in 1999, the chitosan scaffold has a flaky honeycomb structure with the thickness of tens to hundreds of microns, and has good mechanical strength and biological property (Biomaterials, 1999;20: 1133-. Hutch and the like disclose a preparation method of a three-dimensional ordered porous chitosan scaffold material (CN 101366972), which can be used for repairing bone tissues. The microstructure of the scaffold has important influence on the biological performance of the scaffold, and researches show that the nanofiber scaffold has higher specific surface area and is beneficial to cell adhesion and differentiation. But the mechanical strength of the pure nano fiber is lower, and the mechanical load of the stent implantation process cannot be born. In contrast, the micron fiber scaffold has better mechanical properties. Thus, having a microstructure of both nano and micro fibers in the scaffold can overcome the contradiction between biocompatibility and mechanical properties. At present, researchers mostly adopt an electrostatic spinning method to prepare micro-nanofibers, and the electrostatic spinning method can only obtain a two-dimensional film without a special device, and the preparation time is long. The traditional phase separation method for preparing the chitosan three-dimensional tissue engineering scaffold is generally a sheet-shaped honeycomb structure, and the dimension of the chitosan three-dimensional tissue engineering scaffold is usually dozens of micrometers. As the chitosan molecules have stronger intermolecular force and have larger viscosity after being dissolved in acid, the concentration of the chitosan needs to be reduced when the micro-nano fiber structure is obtained, but the prepared scaffold has no mechanical strength and cannot be applied to tissue engineering.
Disclosure of Invention
The invention mainly assumes that a binary solvent system consisting of water and acid in the traditional phase separation preparation method is changed, a novel ternary solvent system consisting of an organic solvent, the water and the acid is added, the intermolecular force of chitosan is changed, and the chitosan is promoted to form micro-nano fibers in the freezing process, so that the chitosan three-dimensional tissue engineering scaffold is obtained.
The preparation method of the chitosan micro-nano fibrous three-dimensional tissue engineering scaffold is specifically as follows.
(1) Preparing a ternary mixed solvent of acetic acid, 1, 4-dioxane and water.
(2) Adding chitosan, and electromagnetically stirring to dissolve for 24 hours.
(3) And (4) after the solution is subjected to sample separation, placing the solution at low temperature for freezing and forming.
(4) Freeze-drying for 48 hours by a freeze dryer to obtain the chitosan micro-nano fibrous three-dimensional tissue engineering scaffold sample.
The sample prepared by the method is in a white foam shape, and the three-dimensional shape of the sample can be adjusted by adopting different containers and sample dividing volumes in sample dividing. The coexistence microstructure of the micro-nano fibers endows the scaffold with good biocompatibility and mechanical strength. The preparation method has simple process and good formability, and compared with the chitosan bracket prepared by the traditional binary solvent, the microstructure of the chitosan bracket is in the form of micro-nano fibers.
Detailed Description
The present invention will be further described with reference to specific examples, but the implementation of the present invention is not limited to these examples.
Example 1: respectively taking 1ml, 10ml and 89ml of acetic acid, 1, 4-dioxane and water, and uniformly mixing. 0.5g of chitosan with the molecular weight of 10 ten thousand is weighed and dissolved for 24 hours by electromagnetic stirring. After the chitosan is completely dissolved, the chitosan solution is sampled into a container, the container is frozen at the temperature of minus 18 ℃ for more than 12 hours for forming, and the three-dimensional tissue engineering scaffold with the chitosan micro-nano fibrous structure can be obtained after the container is frozen in a freeze dryer for 48 hours.
Example 2: respectively taking 10ml, 50ml and 40ml of acetic acid, 1, 4-dioxane and water, and uniformly mixing. 3g of chitosan with the molecular weight of 40 ten thousand is weighed and dissolved for 24 hours by electromagnetic stirring. After the chitosan is completely dissolved, the chitosan solution is divided into samples and put into a container, the samples are frozen and formed under the temperature of 196 ℃ below zero (in liquid nitrogen), and the three-dimensional tissue engineering scaffold with the chitosan micro-nano fibrous structure can be obtained after the samples are frozen and formed in a freeze dryer for 48 hours.
Claims (4)
1. A preparation method of a chitosan nano-micron fibrous three-dimensional tissue engineering scaffold is characterized in that chitosan is dissolved in a ternary mixed solvent of acetic acid, 1, 4-dioxane and water, sample separation is carried out, freezing forming is carried out at a certain temperature, and freeze drying is carried out in a freeze dryer to obtain the chitosan nano-micron fibrous three-dimensional tissue engineering scaffold.
2. The method for preparing a chitosan nano-micron fibrous three-dimensional tissue engineering scaffold according to claim 1, wherein the molecular weight of the chitosan is 10-40 ten thousand, and the mass volume concentration of the chitosan is 0.5-3%.
3. The method for preparing a chitosan nano-micron fibrous three-dimensional tissue engineering scaffold according to claim 1, wherein the volume ratio of acetic acid in the ternary mixed solvent is between 1 and 10%, the volume ratio of 1, 4-dioxane is between 10 and 50%, and the volume ratio of water is between 40 and 89%.
4. The method for preparing a chitosan nano-micron fibrous three-dimensional tissue engineering scaffold according to claim 1, wherein the freezing and forming temperature is between-18 ℃ and-196 ℃.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710433540.2A CN107376012B (en) | 2017-06-09 | 2017-06-09 | Chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710433540.2A CN107376012B (en) | 2017-06-09 | 2017-06-09 | Chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and preparation method thereof |
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| Publication Number | Publication Date |
|---|---|
| CN107376012A CN107376012A (en) | 2017-11-24 |
| CN107376012B true CN107376012B (en) | 2020-03-31 |
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| CN201710433540.2A Expired - Fee Related CN107376012B (en) | 2017-06-09 | 2017-06-09 | Chitosan micro-nano fibrous three-dimensional tissue engineering scaffold and preparation method thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102747453A (en) * | 2012-07-05 | 2012-10-24 | 四川大学 | Porous superfine polymer fiber and preparation method thereof |
| CN103285424A (en) * | 2013-05-27 | 2013-09-11 | 东华大学 | Three-dimensional fiber-based aerogel tissue engineering scaffold and preparation method thereof |
| CN105641744A (en) * | 2016-02-27 | 2016-06-08 | 青岛大学 | Nano-micro multi-scale chitosan and polylactic acid composite scaffold and preparation method thereof |
| WO2017075554A1 (en) * | 2015-10-29 | 2017-05-04 | Golfetto Michael | Methods freeze drying and composite materials |
-
2017
- 2017-06-09 CN CN201710433540.2A patent/CN107376012B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102747453A (en) * | 2012-07-05 | 2012-10-24 | 四川大学 | Porous superfine polymer fiber and preparation method thereof |
| CN103285424A (en) * | 2013-05-27 | 2013-09-11 | 东华大学 | Three-dimensional fiber-based aerogel tissue engineering scaffold and preparation method thereof |
| WO2017075554A1 (en) * | 2015-10-29 | 2017-05-04 | Golfetto Michael | Methods freeze drying and composite materials |
| CN105641744A (en) * | 2016-02-27 | 2016-06-08 | 青岛大学 | Nano-micro multi-scale chitosan and polylactic acid composite scaffold and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 添加剂聚乙二醇对壳聚糖超滤膜结构和性能的影响;刘强等;《膜科学与技术》;20100228;第30卷(第1期);正文第24页左栏第1-3段、实验部分、讨论部分 * |
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| Publication number | Publication date |
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| CN107376012A (en) | 2017-11-24 |
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Granted publication date: 20200331 |