CN114632189A - Elastic porous scaffold and preparation method and application thereof - Google Patents
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- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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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, 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 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 extensive bone injuries, however, these materials have their own drawbacks which are not negligible. Although the autologous bone transplantation is easily accepted by patients, new trauma and pain are caused to the patients, and although the allogeneic bone materials are convenient to obtain, the risks of disease transmission and immune rejection exist, so that the satisfactory repairing effect is difficult to achieve on the aspect 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, but the methods cannot perfectly simulate the natural extracellular matrix structure. With the increasing demand of modern society for bone tissue engineering materials and the increasing demand for the functions of tissue engineering scaffolds, electrostatic spinning, self-assembly and phase separation technologies are in line with the growing demand. 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 may be structurally damaged. The phase separation technology can control the shape, the pore size distribution and the size of 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 polymer 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 scaffolds, 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) performing brittle fracture 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 to 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 step (2), the mass concentration of the short fiber homogenate is 1-50%.
Preferably, 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 1:100-100:1, and the mass concentration of the mixed solution is 0.1-40%.
Preferably, in the step (3), the pore size of the three-dimensional scaffold is 1 to 50 μm, and the porosity is 20 to 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 1-48 h.
Preferably, in the step (4), the mass ratio of the cross-linking agent in the polymer solution of the cross-linking agent to the polymer solution is 1:100-100:1, the low-temperature freezing temperature is (-100) -0 ℃, and the low-temperature freezing time is 0.5-24 h.
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 a use of the elastic porous scaffold, and the application 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 spectrum of an elastic porous scaffold before and after crosslinking according to 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 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 macromolecules as raw materials, prepares a nanofiber membrane through electrostatic spinning, prepares short fiber homogenate by using a homogenizer, mixes the short fiber homogenate with a macromolecule solution, thereby carries out external casting treatment, and prepares a three-dimensional scaffold with a porous structure through freeze drying, and finally obtains an elastic porous scaffold material with good elasticity and a communicated pore structure in the interior, 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 a 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), while the common macroporous stent has the advantages that the pores collapse after the compression and cannot return to the original shape, such as the original thickness of 1cm, and the thickness of the 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 readily apparent to those skilled in the art that various modifications to these embodiments and the generic principles defined 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 should appreciate that many modifications and variations are possible in light of the above teaching without departing from the scope of the invention.
Claims (10)
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) 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, soaking, washing, and freeze-drying to obtain the elastic porous scaffold.
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 more than one of synthetic polymer and natural polymer;
preferably, the synthetic polymer is selected from more than one of polylactic acid, polyglycolic acid, polycaprolactone and polyethylene glycol;
preferably, the natural polymer is selected from more than one of collagen, hyaluronic acid, chitosan, silk fibroin and sodium alginate.
3. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (1), the solvent is more than one selected from hexafluoroisopropanol, trifluoroacetic acid, dichloromethane, trichloromethane, tetrahydrofuran, dimethyl sulfoxide and ethyl acetate; and/or the presence of a gas in the atmosphere,
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 8-20 cm.
4. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (2), the brittle fracture solution solvent is selected from one or more mixed solutions of ethanol, polyacrylamide and tert-butyl alcohol and water; and/or the presence of a gas in the gas,
in the step (2), the mass concentration of the short fiber homogenate is 1-50%.
5. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: 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 1:100-100:1, and the mass concentration of the mixed solution is 0.1-40%; and/or the presence of a gas in the atmosphere,
in the step (3), the aperture of the three-dimensional scaffold is 1-50 μm, and the porosity is 20-90%.
6. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (4), the cross-linking agent is selected from more than one of genipin and gallic acid; and/or the presence of a gas in the gas,
in the step (4), the cross-linking time of the three-dimensional scaffold is 1-48 h; and/or the presence of a gas in the gas,
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 1:100-100: 1; the low-temperature freezing temperature is (-100) -0 ℃, and the low-temperature freezing time is 0.5-24 h.
7. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (3) and the step (4), the polymer substance in the polymer solution is selected from more than one of polyvinyl alcohol, sodium dodecyl benzene sulfonate, polyurethane, polylactic acid-glycolic acid copolymer, silk fibroin, polyethylene glycol, collagen and gelatin; and/or the presence of a gas in the gas,
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.
8. The method of preparing an elastic porous scaffold according to claim 1, characterized in that: in the step (4), the aperture of the elastic porous bracket is 1-50 μm, and the porosity is 20-90%.
9. An elastic porous scaffold characterized by: which is obtained by the production method according to any one of claims 1 to 8.
10. Use of the elastic porous scaffold according to claim 9 in bone tissue engineering and regenerative medicine.
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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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Cited By (3)
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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 |
WO2023179422A1 (en) * | 2022-03-25 | 2023-09-28 | 上海工程技术大学 | Elastic porous scaffold, preparation method therefor and use thereof |
CN115990289B (en) * | 2023-01-09 | 2024-05-31 | 西岭(镇江)医疗科技有限公司 | Preparation method of incomplete decalcification osteoinductive material |
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CN105107022A (en) * | 2015-09-21 | 2015-12-02 | 东华大学 | Preparation method for nanofiber porous scaffold having compression elasticity in wet state |
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 |
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WO2023179422A1 (en) * | 2022-03-25 | 2023-09-28 | 上海工程技术大学 | Elastic porous scaffold, preparation method therefor and use thereof |
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 |
CN115990289B (en) * | 2023-01-09 | 2024-05-31 | 西岭(镇江)医疗科技有限公司 | Preparation method of incomplete decalcification osteoinductive material |
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