CN103690991B - A kind of preparation method of cartilage repairing scaffold material - Google Patents
A kind of preparation method of cartilage repairing scaffold material Download PDFInfo
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- CN103690991B CN103690991B CN201310690006.1A CN201310690006A CN103690991B CN 103690991 B CN103690991 B CN 103690991B CN 201310690006 A CN201310690006 A CN 201310690006A CN 103690991 B CN103690991 B CN 103690991B
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Abstract
The present invention relates to tissue engineering bracket technical field, especially a kind of preparation method of cartilage repairing scaffold material is related to, first by the chitosan nano fiber of electrostatic spinning preparation containing hydroxyapatite, then hyaluronic acid encapsulation is used, by the experimental technique of lyophilizing, the cartilage repairing scaffold material of preparation.The preparation method of a kind of cartilage repairing scaffold material of the present invention, the cartilage repairing scaffold material obtained has larger specific surface area, be conducive to growth and the propagation of chondrocyte, hydroxyapatite is as the main component in human bone, for a large amount of necessary raw material that the growth of chondrocyte provides, hyaluronic acid is the main component of cartilage liquid, be conducive to giving chondrocyte one good growing environment, simultaneously hyaluronic acid can with chitosan compound, nanometer hydroxyapatite is encapsulated in chitin fiber, form a kind of slow-releasing system of hydroxyapatite, somatomedin can promote chondrocyte fast breeding.
Description
Technical field
The present invention relates to tissue engineering bracket technical field, especially relate to a kind of preparation method of cartilage repairing scaffold material.
Background technology
Articular cartilage is one tissue comparatively special in human body, and it does not have blood vessel, nerve and lymphocyte, and thus self-reparing capability is very poor.At present Therapeutic Method traditional clinically mainly contains attrition, Cartilage transplantation and autogenous cell transplantation art etc. under microfrature, subchondral bone Drilling, cartilage, but these methods exist many deficiencies, and can bring secondary insult to patient.Also do not have a kind of method successfully can substitute damage location and reach the object of regenerating bone or cartilage at present.
The cartilage repairing scaffold material of current use can be divided into natural scaffold materials and the large class of man-made support material two substantially.Natural scaffold materials derives from naturally occurring macromolecular material in animal and plant or human body, has good biocompatibility, organizes affinity.Man-made support material is the material that research is more at present, mainly contains polyglycolic acid, polylactic acid and condensing model etc., can adjust its physics, chemistry, biomechanics and degradation property as required, easily shape, produce reproducible.Current natural scaffold materials is mainly by being processed into the form of hydrogel as cartilaginous tissue recovery support using materials such as natural polymers, but the poor performance of the repair of cartilage support breathability made of hydrogel and cell proliferation and growth.
Summary of the invention
The technical problem to be solved in the present invention is: in order to overcome the problems of the prior art, provides a kind of preparation method of cartilage repairing scaffold material.
The technical solution adopted for the present invention to solve the technical problems is: a kind of preparation method of cartilage repairing scaffold material, comprises the following steps:
Step 1) preparation of chitosan solution: preparation mass concentration is the acetic acid aqueous solution of 1-10wt% chitosan, it is fully stirred, obtains the chitosan solution of transparent and homogeneous;
Step 2) preparation of chitosan solution of hydroxyapatite: in step 1) in add nanometer hydroxyapatite in chitosan solution, its mass concentration is 10-25wt%, make it fully stir, obtain the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Step 3) water solublity synthesis macromolecular solution preparation: preparation mass concentration be that 1-10wt% water solublity synthesizes high molecular aqueous solution, it fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution;
Step 4) preparation of electrostatic spinning liquid: by step 2) in the chitosan solution of hydroxyapatite and step 3) in water solublity to synthesize high molecular aqueous solution be the ratio mix homogeneously of 5-9:1 by volume;
Step 5) method of electrostatic spinning prepares chitosan nano fiber: by step 4) in the solution of preparation join in solution supplying device, spinning voltage 10 ~ 20kV is set, spinning distance 10 ~ 20cm between spinning mouth and receiving system, flow speed control 0.1 ~ 1.2ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 1-5wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Step 6) purification of chitosan nano fiber: by step 5) nanofiber that obtains soaks 4-12 hour in deionized water, remove the water solublity synthesis macromolecule in nanofiber, dry and obtain chitosan nano fiber;
Step 7) preparation of cartilage repairing scaffold material: by step 6) in chitosan nano fiber to be immersed in mass concentration be in the hyaluronic acid aqueous solution of 1-10wt%, by freezer dryer lyophilizing, can cartilage repairing scaffold material be obtained.
In order to improve the spinning property of chitosan, described water solublity synthesis macromolecule is one or more in PVP, PVA, PEO and PEG, effectively can promote the formation of chitosan nano fiber.
Its body ground, the mass concentration of described acetic acid aqueous solution is 1-5wt%.
In order to accelerate the growth of cell on cartilage repairing scaffold material and propagation, described hyaluronic acid solution contains somatomedin.
As preferably, the content of described somatomedin is the 10-30% of hyaluronic acid quality.
Particularly, described hyaluronic weight average molecular weight is M
w=5000 ~ 2000000g/mol.
The invention has the beneficial effects as follows: the preparation method of a kind of cartilage repairing scaffold material of the present invention, the cartilage repairing scaffold material obtained has larger specific surface area, be conducive to growth and the propagation of chondrocyte, hydroxyapatite is as the main component in human bone, for a large amount of necessary raw material that the growth of chondrocyte provides, hyaluronic acid is the main component of cartilage liquid, be conducive to giving chondrocyte one good growing environment, simultaneously hyaluronic acid can with chitosan compound, nanometer hydroxyapatite is encapsulated in chitin fiber, form a kind of slow-releasing system of hydroxyapatite, somatomedin can promote chondrocyte fast breeding.
Detailed description of the invention
Below in conjunction with specific embodiment, further the present invention is set forth, should be understood that and quote embodiment only for illustration of the present invention, and be not used in and limit the scope of the invention.
Embodiment 1
A preparation method for cartilage repairing scaffold material, comprises the following steps:
Preparation mass concentration is the acetic acid aqueous solution of 1wt% chitosan, and the mass concentration of described acetic acid aqueous solution is 5wt%, it is fully stirred, and obtains the chitosan solution of transparent and homogeneous;
In chitosan solution, add nanometer hydroxyapatite, make its mass concentration be 10wt%, it is fully stirred, obtain the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Preparation mass concentration is the aqueous solution of 10wt%PVP, and it fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution;
By the ratio mix homogeneously that the aqueous solution of the chitosan solution containing hydroxyapatite and PVP is 5:1 by volume;
Mixed solution is joined in solution supplying device, spinning voltage 10kV is set, spinning distance 10cm between spinning mouth and receiving system, flow speed control 0.1ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 1wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Nanofiber is soaked 4 hours in deionized water, removes the PVP in nanofiber, dry and obtain chitosan nano fiber;
Chitosan nano fiber is immersed in mass concentration is 1wt%, weight average molecular weight is M
win the hyaluronic acid aqueous solution of=2000000g/mol, described hyaluronic acid solution contains somatomedin, and the content of somatomedin is 10% of hyaluronic acid quality, by freezer dryer lyophilizing, can obtain cartilage repairing scaffold material.
Embodiment 2
A preparation method for cartilage repairing scaffold material, comprises the following steps:
Preparation mass concentration is the acetic acid aqueous solution of 10wt% chitosan, and the mass concentration of described acetic acid aqueous solution is 1wt%, it is fully stirred, and obtains the chitosan solution of transparent and homogeneous;
In chitosan solution, add nanometer hydroxyapatite, make its mass concentration be 25wt%, it is fully stirred, obtain the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Preparation mass concentration is the aqueous solution of 1wt%PEO, and it fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution;
By the ratio mix homogeneously that the aqueous solution of the chitosan solution containing hydroxyapatite and PEO is 9:1 by volume;
Mixed solution is joined in solution supplying device, spinning voltage 20kV is set, spinning distance 20cm between spinning mouth and receiving system, flow speed control 1.2ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 5wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Nanofiber is soaked 12 hours in deionized water, removes the PEO in nanofiber, dry and obtain chitosan nano fiber;
Chitosan nano fiber is immersed in mass concentration is 10wt%, weight average molecular weight is M
win the hyaluronic acid aqueous solution of=5000g/mol, described hyaluronic acid solution contains somatomedin, and the content of somatomedin is 30% of hyaluronic acid quality, by freezer dryer lyophilizing, can obtain cartilage repairing scaffold material.
Embodiment 3
A preparation method for cartilage repairing scaffold material, comprises the following steps:
Preparation mass concentration is the acetic acid aqueous solution of 4wt% chitosan, and the mass concentration of described acetic acid aqueous solution is 4wt%, it is fully stirred, and obtains the chitosan solution of transparent and homogeneous;
In chitosan solution, add nanometer hydroxyapatite, make its mass concentration be 15wt%, it is fully stirred, obtain the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Preparation mass concentration is the aqueous solution of 3wt%PEG, and it fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution;
By the ratio mix homogeneously that the aqueous solution of the chitosan solution containing hydroxyapatite and PEG is 7:1 by volume;
Mixed solution is joined in solution supplying device, spinning voltage 16kV is set, spinning distance 10cm between spinning mouth and receiving system, flow speed control 0.9ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 2.5wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Nanofiber is soaked 6 hours in deionized water, removes the PEG in nanofiber, dry and obtain chitosan nano fiber;
Chitosan nano fiber is immersed in mass concentration is 4wt%, weight average molecular weight is M
win the hyaluronic acid aqueous solution of=100000g/mol, described hyaluronic acid solution contains somatomedin, and the content of somatomedin is 18% of hyaluronic acid quality, by freezer dryer lyophilizing, can obtain cartilage repairing scaffold material.
Embodiment 4
A preparation method for cartilage repairing scaffold material, comprises the following steps:
Preparation mass concentration is the acetic acid aqueous solution of 3wt% chitosan, and the mass concentration of described acetic acid aqueous solution is 3.5wt%, it is fully stirred, and obtains the chitosan solution of transparent and homogeneous;
In chitosan solution, add nanometer hydroxyapatite, make its mass concentration be 22wt%, it is fully stirred, obtain the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Preparation mass concentration is that 10wt% water solublity synthesizes high molecular aqueous solution, and it fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution;
By the ratio mix homogeneously that the aqueous solution of the chitosan solution containing hydroxyapatite and PVA is 5-9:1 by volume;
Mixed solution is joined in solution supplying device, spinning voltage 20kV is set, spinning distance 18cm between spinning mouth and receiving system, flow speed control 1.2ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 5wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Nanofiber is soaked 8 hours in deionized water, removes the PVA in nanofiber, dry and obtain chitosan nano fiber;
Chitosan nano fiber is immersed in mass concentration is 4wt%, weight average molecular weight is M
win the hyaluronic acid aqueous solution of=200000g/mol, described hyaluronic acid solution contains somatomedin, and the content of somatomedin is 10% of hyaluronic acid quality, by freezer dryer lyophilizing, can obtain cartilage repairing scaffold material.
Embodiment 5
A preparation method for cartilage repairing scaffold material, comprises the following steps:
Preparation mass concentration is the acetic acid aqueous solution of 5wt% chitosan, and the mass concentration of described acetic acid aqueous solution is 2wt%, it is fully stirred, and obtains the chitosan solution of transparent and homogeneous;
In chitosan solution, add nanometer hydroxyapatite, make its mass concentration be 13wt%, it is fully stirred, obtain the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Preparation mass concentration is that 4wt% water solublity synthesizes high molecular aqueous solution, and it fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution;
By the ratio mix homogeneously that the aqueous solution of the chitosan solution containing hydroxyapatite and PVA, PVP is 8:1 by volume;
Mixed solution is joined in solution supplying device, spinning voltage 12kV is set, spinning distance 11cm between spinning mouth and receiving system, flow speed control 1.1ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 2wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Nanofiber is soaked 4 hours in deionized water, removes PVA, PVP in nanofiber, dry and obtain chitosan nano fiber;
Chitosan nano fiber is immersed in mass concentration is 4wt%, weight average molecular weight is M
win the hyaluronic acid aqueous solution of=10000g/mol, described hyaluronic acid solution contains somatomedin, and the content of somatomedin is 30% of hyaluronic acid quality, by freezer dryer lyophilizing, can obtain cartilage repairing scaffold material.
With above-mentioned according to desirable embodiment of the present invention for enlightenment, by above-mentioned description, relevant staff in the scope not departing from this invention technological thought, can carry out various change and amendment completely.The technical scope of this invention is not limited to the content in description, must determine its technical scope according to right.
Claims (5)
1. a preparation method for cartilage repairing scaffold material, is characterized in that comprising the following steps:
Step 1) preparation of chitosan solution: preparation mass concentration is the acetic acid aqueous solution of 1-10wt% chitosan, it is fully stirred, obtains the chitosan solution of transparent and homogeneous;
Step 2) preparation of chitosan solution of hydroxyapatite: in step 1) in add nanometer hydroxyapatite in chitosan solution, its mass concentration is made to be 10-25wt%, it is fully stirred, obtains the chitosan solution of the hydroxyapatite of transparent and homogeneous;
Step 3) water solublity synthesis macromolecular solution preparation: preparation mass concentration be that 1-10wt% water solublity synthesizes high molecular aqueous solution, it is fully stirred, the water solublity obtaining transparent and homogeneous synthesizes high molecular aqueous solution, and described water solublity synthesis macromolecule is one or more in PVP, PVA, PEO and PEG;
Step 4) preparation of electrostatic spinning liquid: by step 2) in the chitosan solution of hydroxyapatite and step 3) in water solublity synthesize high molecular aqueous solution by volume for 5-9: 1 ratio mix homogeneously;
Step 5) method of electrostatic spinning prepares chitosan nano fiber: by step 4) in the solution of preparation join in solution supplying device, spinning voltage 10 ~ 20kV is set, spinning distance 10 ~ 20cm between spinning mouth and receiving system, flow speed control 0.1 ~ 1.2ml/h, the genipin alcoholic solution of receiving system to be the mass concentration of ground connection be 1-5wt%, starting drive carries out electrostatic spinning, in receiving system, collect chitosan nano fiber;
Step 6) purification of chitosan nano fiber: by step 5) nanofiber that obtains soaks 4-12 hour in deionized water, remove the water solublity synthesis macromolecule in nanofiber, dry and obtain chitosan nano fiber;
Step 7) preparation of cartilage repairing scaffold material: by step 6) in chitosan nano fiber to be immersed in mass concentration be in the hyaluronic acid aqueous solution of 1-10wt%, by freezer dryer lyophilizing, can cartilage repairing scaffold material be obtained.
2. the preparation method of cartilage repairing scaffold material as claimed in claim 1, is characterized in that: the mass concentration of described acetic acid aqueous solution is 1-5wt%.
3. the preparation method of cartilage repairing scaffold material as claimed in claim 1 or 2, is characterized in that: described hyaluronic acid solution contains somatomedin.
4. the preparation method of cartilage repairing scaffold material as claimed in claim 3, is characterized in that: the content of described somatomedin is the 10-30% of hyaluronic acid quality.
5. the preparation method of cartilage repairing scaffold material as claimed in claim 1, is characterized in that: described hyaluronic weight average molecular weight is M
w=5000 ~ 2000000g/mol.
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| CN104998302B (en) * | 2015-08-31 | 2017-11-24 | 中原工学院 | It is a kind of using oriented nanofibers felt as nanometer cartilage repair material of skeleton and preparation method thereof |
| CN105311676B (en) * | 2015-11-10 | 2018-09-11 | 北京大清生物技术股份有限公司 | A kind of biologically active hard tissue engineering timbering material and preparation method thereof |
| CN107802888A (en) * | 2017-10-31 | 2018-03-16 | 无锡中科光远生物材料有限公司 | A kind of preparation method of the nano fiber scaffold of promotion regenerating bone or cartilage |
| CN107653498A (en) * | 2017-11-10 | 2018-02-02 | 安徽翰联纺织有限公司 | Electrostatic spinning prepares the method for doping antibacterial granule fiber, electrostatic spinning machine |
| KR102170467B1 (en) * | 2018-04-16 | 2020-10-28 | 성균관대학교산학협력단 | Cell scaffold with fiber bundle structure and method of manufacturing the same |
| CN109745582B (en) * | 2019-03-11 | 2021-11-19 | 西南科技大学 | Preparation method of bletilla striata polysaccharide composite cytoskeleton |
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| CN111235886A (en) * | 2020-03-23 | 2020-06-05 | 动之医学技术(上海)有限公司 | Preparation method of chitosan-based composite fiber and tissue engineering scaffold |
| CN113045760B (en) * | 2021-04-12 | 2022-03-25 | 华南理工大学 | Polymer brush based on hyaluronic acid and alpha-D-galactose and preparation method and application thereof |
| CN115678233B (en) * | 2023-01-03 | 2023-03-31 | 北京德益达美医疗科技有限公司 | Toughened absorbable composite material and preparation method thereof |
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