CN114632189B - Elastic porous scaffold and preparation method and application thereof - Google Patents
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- 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
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- 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
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
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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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
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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
- A61L27/26—Mixtures of macromolecular compounds
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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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
Abstract
The invention provides an elastic porous scaffold and a preparation method and application thereof, wherein the preparation method comprises the following steps: dissolving a high molecular polymer in a solvent, and performing electrostatic spinning to obtain a nanofiber membrane; performing brittle fracture in a brittle fracture solution solvent to form short fiber homogenate; mixing the solution with a polymer solution to form a mixed solution, and then pouring the mixed solution into a mold for freeze drying to obtain a three-dimensional scaffold; immersing into polymer solution with cross-linking agent, freezing at low temperature, and freeze-drying; according to the invention, the electrostatic spinning technology and the freeze drying technology are combined, the prepared scaffold has a nanofiber structure, and the internal communicated pore channel is beneficial to cell proliferation and adhesion; the stent after the outer casting treatment can recover the original shape when the strain reaches 80% in a wet state, has good elastic property, high porosity and high water absorption performance, and can be widely applied to the aspects of bone tissue engineering and regenerative medicine.
Description
Technical Field
The invention belongs to the field of biomaterial bone tissue engineering and regenerative medicine, and particularly relates to an elastic porous scaffold and a preparation method and application thereof.
Background
The transplantation of autologous or allogeneic bone material in vivo has been the method used to repair large areas of bone damage, however, these materials have their own drawbacks that cannot be ignored. Although the autologous bone transplantation is easily accepted by patients, new trauma and pain are caused to the patients, and although the allogenic bone materials are convenient to obtain, the risks of disease transmission and immune rejection exist, so that a satisfactory repair effect is difficult to achieve in terms of biological safety. Bone tissue engineering and regenerative medicine have been developed against this background, and new expectations are being placed on bone repair.
The traditional three-dimensional scaffold can be prepared by a plurality of methods, such as a hot-pressing forming method, a solvent casting method, a rapid forming method and the like, and the tissue engineering scaffold can be prepared by the methods, but the methods cannot perfectly simulate a natural extracellular matrix structure. With the increasing demand of modern society for bone tissue engineering materials and the higher and higher requirements for the functions of tissue engineering scaffolds, electrostatic spinning, self-assembly and phase separation technologies come into force. The fiber membrane prepared by the electrostatic spinning technology is formed by stacking multiple layers of fibers, has poor mechanical property and cannot be well applied to bone defects. The membranes or scaffolds prepared by self-assembly techniques may have insufficient stability and structural failure. The phase separation technology can control the shape, the pore size distribution and the size of the pores of the bracket, but the mechanical property of the bracket is poor. Therefore, it is important to select a suitable method for preparing a scaffold with elastic properties and an internally communicated porous structure.
Degradable synthetic high molecular compounds such as polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), polyethylene glycol and the like, have the advantages of excellent biocompatibility, no toxicity, good encapsulation and film-forming properties, good osteoinductive potential and the like, are commonly used for preparing microspheres, nanoparticles and tissue engineering supports, and are widely applied to the field of bone tissue engineering.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide an elastic porous scaffold and a preparation method and application thereof. The scaffold has high porosity and large aperture, can simulate the structure of natural extracellular matrix, and can be widely applied to bone tissue engineering and regenerative medicine.
In order to achieve the above purpose, the solution of the invention is as follows:
in a first aspect, the present invention provides a method for preparing an elastic porous scaffold, comprising the steps of:
(1) Dissolving a high polymer material in a solvent, stirring to obtain a spinning solution, and performing electrostatic spinning to obtain a nanofiber membrane;
(2) Brittle fracture is carried out on the nanofiber membrane in a brittle fracture solution solvent to form short fiber homogenate;
(3) Fully mixing the short fiber homogenate with the polymer solution to form a mixed solution, and then pouring the mixed solution into a mould for freeze drying to obtain a three-dimensional scaffold;
(4) And immersing the three-dimensional scaffold into a high molecular solution with a cross-linking agent, freezing the cross-linked scaffold at a low temperature, immersing, washing, and freeze-drying to obtain the elastic porous scaffold.
Preferably, in the step (1), the polymer material is one or more of a synthetic polymer and a natural polymer.
Preferably, the synthetic polymer is selected from one or more of polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), and polyethylene glycol.
Preferably, the natural polymer is selected from more than one of collagen, hyaluronic acid, chitosan, silk fibroin and sodium alginate.
Preferably, in the step (1), the solvent is one or more selected from hexafluoroisopropanol, trifluoroacetic acid, dichloromethane, chloroform, tetrahydrofuran, dimethyl sulfoxide and ethyl acetate.
Preferably, in the step (1), the electrostatic spinning process is as follows: adding the spinning solution into an injector, installing the injector on a propulsion pump, connecting high pressure, setting the voltage of a spinning machine to be 1-12KV, the propulsion speed to be 0.5-4mL/h, and the distance from an aluminum foil receiver to be 8-20cm, and carrying out electrostatic spinning.
Preferably, in the step (2), the brittle fracture solvent is selected from one or more mixed solutions of ethanol, polyacrylamide and tertiary butanol and water.
Preferably, in the step (2), the mass concentration of the short fiber homogenate is 1 to 50%.
Preferably, in the step (3), the mass concentration of the polymer solution is 1 to 50%, the volume ratio of the short fiber homogenate to the polymer solution is 1.
Preferably, in the step (3), the pore size of the three-dimensional scaffold is 1-50 μm, and the porosity is 20-90%.
Preferably, in the step (4), the cross-linking agent is a naturally extracted green cross-linking agent, and the cross-linking agent is selected from more than one of genipin and gallic acid.
Preferably, in the step (4), the cross-linking time of the three-dimensional scaffold is 1-48h.
Preferably, in the step (4), the mass ratio of the cross-linking agent to the polymer solution in the polymer solution of the cross-linking agent is 1.
Preferably, in the step (3) and the step (4), the polymer in the polymer solution is selected from one or more of polyvinyl alcohol (PVA), sodium Dodecyl Benzene Sulfonate (SDBS), polyurethane, polylactic-co-glycolic acid (PLGA), silk fibroin, polyethylene glycol, collagen and gelatin.
Preferably, in the step (3) and the step (4), the solvent in the polymer solution is selected from one or more of deionized water, dimethyl sulfoxide, tetrahydrofuran and 1, 4-dioxane.
In the step (3), the dissolving temperature of the high molecular substance in the high molecular solution is 20-110 ℃.
Preferably, in the step (4), the elastic porous scaffold has a pore size of 1 to 50 μm and a porosity of 20 to 90%.
In a second aspect, the present invention provides an elastic porous scaffold obtained by the above-described preparation method.
In a third aspect, the invention provides the use of the elastic porous scaffold, and the use of the elastic porous scaffold in bone tissue engineering and regenerative medicine.
Due to the adoption of the scheme, the invention has the beneficial effects that:
according to the invention, by combining the electrostatic spinning technology and the freeze drying technology, the prepared scaffold has a nanofiber structure, and the pore passages communicated with the inside of the scaffold are beneficial to cell proliferation and adhesion; the stent after the outer casting treatment can recover the original shape when the strain reaches 80% in a wet state, has good elastic property, high porosity and high water absorption performance, and can be widely applied to the aspects of bone tissue engineering and regenerative medicine.
Drawings
FIG. 1 is a scanning electron micrograph of an elastic porous scaffold according to example 1 of the present invention.
FIG. 2 is a FT-IR analysis of the elastic porous scaffold before and after crosslinking of example 2 of the present invention.
FIG. 3 is a graph of compressive stress-strain curves for the resilient porous scaffold of example 2 of the present invention.
Fig. 4 is a water absorption rate graph of the elastic porous support according to example 2 of the present invention.
Detailed Description
The invention provides an elastic porous scaffold and a preparation method and application thereof.
The invention mainly takes high polymer as raw material, prepares nano fiber membrane through electrostatic spinning, then uses a refiner to prepare short fiber homogenate, mixes with high polymer solution, thereby carries out external casting treatment, and prepares a three-dimensional bracket with porous structure through freeze drying, and finally obtains an elastic porous bracket material with good elasticity and communicated pore structure inside, which is beneficial to the proliferation and adhesion of cells and can be widely applied to the reverse side of bone tissue engineering and regenerative medicine.
The invention is further illustrated by the following figures and examples.
Example 1:
weighing 1g of polyglycolic acid and silk fibroin, dissolving in 10mL of hexafluoroisopropanol, uniformly stirring, and performing electrostatic spinning to obtain the uniformly distributed nanofiber membrane. The prepared nanofiber membrane was cut into pieces and weighed. The nanofiber membrane was brittle in polyacrylamide using a homogenizer to make a short fiber homogenate. Dissolving polyvinyl alcohol (PVA) in water at 90 ℃ (the mass concentration of the PVA is 2.5%), uniformly mixing the polyvinyl alcohol (PVA) with the short fiber homogenate, carrying out ultrasonic treatment, pouring the mixed solution after ultrasonic treatment into a mold, pre-freezing for 1h in a refrigerator at-80 ℃, and then carrying out freeze drying to obtain the three-dimensional scaffold. And soaking the obtained three-dimensional scaffold in a genipin/PVA solution for crosslinking for 24h, freezing for 1h at-20 ℃, then washing for 3 times by using ice deionized water, transferring to a required mould, and obtaining the elastic porous scaffold by freeze drying. Fig. 1 shows the micro-topography of the elastic scaffold.
Example 2:
weighing the components in a mass ratio of 2:1, dissolving the glycolic acid copolymer and the collagen in hexafluoroisopropanol to prepare a solution with the mass concentration of 12%, uniformly stirring, and performing electrostatic spinning to obtain the uniformly distributed nanofiber membrane. The prepared nanofiber membrane was cut into pieces and weighed. The nanofiber membrane was brittle in t-butanol using a homogenizer to a short fiber homogenate of 5% mass concentration. Preparing glycerol into a glycerol/water solution with the mass concentration of 10%, uniformly mixing the glycerol/water solution with the short fiber homogenate, carrying out ultrasonic treatment, pouring the ultrasonic solution into a mold, pre-freezing the mold for 2 hours in a refrigerator with the temperature of-80 ℃, and then carrying out freeze drying to obtain the three-dimensional scaffold. And soaking the obtained three-dimensional scaffold in a gallic acid/PVA solution for crosslinking for 24h, freezing for 1h at the temperature of-20 ℃, then washing for 3 times by using ice deionized water, transferring to a required mould, and obtaining the elastic porous scaffold by freeze drying. The water absorption rate of the stent in water can reach 1200%, and the stent can be restored to the original state and has good elastic performance when the compressive strain reaches 80% in a wet state (as shown in fig. 3, the abscissa is the strain, the ordinate is the stress, and the stress and the strain return to the starting point after the compression is finished, so that the stent is proved to be completely rebounded and has good elasticity), whereas the pores of the common macroporous stent collapse after the compression, and cannot return to the original shape, such as the original thickness of 1cm, and the thickness of the common macroporous stent is only 0.3cm after the compression is finished. The stent prepared by the present invention can return to the original position after being compressed, and thus has good elastic properties, porosity and high water absorption properties (as shown in fig. 2 to 4).
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. It will be apparent to those skilled in the art that various modifications to these embodiments can be readily made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments. Those skilled in the art, having the benefit of the teachings of this invention, will appreciate numerous modifications and variations there from without departing from the scope of the invention as defined by the appended claims.
Claims (14)
1. A preparation method of an elastic porous scaffold is characterized by comprising the following steps: which comprises the following steps:
(1) Dissolving a high polymer material in a solvent, stirring to obtain a spinning solution, and performing electrostatic spinning to obtain a nanofiber membrane;
(2) Brittle fracture is carried out on the nanofiber membrane in a brittle fracture solution solvent to form short fiber homogenate;
(3) Fully mixing the short fiber homogenate with a high molecular solution to form a mixed solution, and then pouring the mixed solution into a mould for freeze drying to obtain a three-dimensional scaffold;
(4) Immersing the three-dimensional scaffold into a high molecular solution with a cross-linking agent, freezing the cross-linked scaffold at a low temperature, soaking, washing, and freeze-drying to obtain an elastic porous scaffold;
in the step (2), the brittle fracture solution solvent is selected from one or more mixed solutions of ethanol, polyacrylamide and water;
in the step (4), the cross-linking agent is selected from more than one of genipin and gallic acid;
in the step (3) and the step (4), the polymer in the polymer solution is selected from more than one of polyvinyl alcohol, sodium dodecyl benzene sulfonate and polyurethane;
in the step (4), the aperture of the elastic porous scaffold is 1-50 μm, and the porosity is 20-90%.
2. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (1), the polymer material is selected from one or more of synthetic polymers and natural polymers.
3. The method of preparing an elastic porous scaffold according to claim 2, characterized in that: the synthetic polymer is selected from more than one of polylactic acid, polyglycolic acid, polycaprolactone and polyethylene glycol.
4. The method of preparing an elastic porous scaffold according to claim 2, characterized in that: the natural polymer is selected from more than one of collagen, hyaluronic acid, chitosan, silk fibroin and sodium alginate.
5. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (1), the solvent is selected from more than one of hexafluoroisopropanol, trifluoroacetic acid, dichloromethane, trichloromethane, tetrahydrofuran, dimethyl sulfoxide and ethyl acetate.
6. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (1), the electrostatic spinning parameters are as follows: the voltage of the spinning machine is 1-12KV, the advancing speed is 0.5-4mL/h, and the receiver distance is 8-20cm.
7. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (2), the mass concentration of the short fiber homogenate is 1-50%.
8. The method for preparing an elastic porous scaffold according to claim 1, wherein: in the step (3), the mass concentration of the polymer solution is 1-50%, the volume ratio of the short fiber homogenate to the polymer solution is 1.
9. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (3), the aperture of the three-dimensional scaffold is 1-50 μm, and the porosity is 20-90%.
10. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (4), the cross-linking time of the three-dimensional scaffold is 1-48h.
11. The method for preparing an elastic porous scaffold according to claim 1, wherein: in the step (4), the mass ratio of the cross-linking agent to the polymer solution in the polymer solution of the cross-linking agent is 1; the low-temperature freezing temperature is (-100) -0 ℃, and the low-temperature freezing time is 0.5-24h.
12. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (3) and the step (4), the solvent in the polymer solution is selected from more than one of deionized water, dimethyl sulfoxide, tetrahydrofuran and 1, 4-dioxane.
13. An elastic porous scaffold, comprising: obtained by the process according to any one of claims 1 to 12.
14. Use of the elastic porous scaffold of claim 13 in the preparation of bone tissue engineering and regenerative medical materials.
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| CN202210301368.6A CN114632189B (en) | 2022-03-25 | 2022-03-25 | Elastic porous scaffold and preparation method and application thereof |
| PCT/CN2023/081587 WO2023179422A1 (en) | 2022-03-25 | 2023-03-15 | Elastic porous scaffold, preparation method therefor and use thereof |
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| CN115990289A (en) * | 2023-01-09 | 2023-04-21 | 西岭(镇江)医疗科技有限公司 | Preparation method of incomplete decalcification osteoinductive material |
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| WO2009154344A1 (en) * | 2008-06-18 | 2009-12-23 | 주식회사 바이오폴 | Preparation method for natural porous hyaluronic acid-collagen polymer support for tissue repair |
| CN105107022A (en) * | 2015-09-21 | 2015-12-02 | 东华大学 | Preparation method for nanofiber porous scaffold having compression elasticity in wet state |
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| CN107185038A (en) * | 2017-05-19 | 2017-09-22 | 广州市朴道联信生物科技有限公司 | A kind of surface immobilized modified cornea repair material and preparation method thereof |
| WO2019209762A1 (en) * | 2018-04-23 | 2019-10-31 | Board Of Regents Of The University Of Nebraska | Nanofiber microspheres and methods of use thereof |
| CN109836595A (en) * | 2019-01-28 | 2019-06-04 | 北京航空航天大学 | A kind of bionical cartilage splices the preparation method of the double-deck hydrogel doped with ordered magnetism nano short fiber |
| CN110538006B (en) * | 2019-08-13 | 2022-06-17 | 上海亚朋生物技术有限公司 | Manufacturing method of fiber-reinforced three-dimensional printing cartilage acellular matrix scaffold |
| CN112064193A (en) * | 2020-08-28 | 2020-12-11 | 华东理工大学 | Preparation method of cellulose diacetate based three-dimensional scaffold with antibacterial and biocompatibility functions |
| CN113750291A (en) * | 2021-10-18 | 2021-12-07 | 诺一迈尔(苏州)生命科技有限公司 | Articular cartilage scaffold and preparation method thereof |
| CN114632189B (en) * | 2022-03-25 | 2023-02-10 | 上海工程技术大学 | Elastic porous scaffold and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2009154344A1 (en) * | 2008-06-18 | 2009-12-23 | 주식회사 바이오폴 | Preparation method for natural porous hyaluronic acid-collagen polymer support for tissue repair |
| CN105107022A (en) * | 2015-09-21 | 2015-12-02 | 东华大学 | Preparation method for nanofiber porous scaffold having compression elasticity in wet state |
| JP2019042280A (en) * | 2017-09-05 | 2019-03-22 | 国立大学法人京都大学 | Elastic fibrous tissue regeneration substrate and method for producing same |
| CN107715174A (en) * | 2017-09-30 | 2018-02-23 | 清华大学 | A kind of bionical tissue engineering bracket containing micropore and nanofiber composite construction and preparation method thereof |
| CN110975009A (en) * | 2019-12-17 | 2020-04-10 | 东南大学 | Preparation method of electrostatic spinning fiber scaffold material |
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