CN109667059B - Method for preparing silk fibroin biological tissue engineering scaffold by solvent spraying - Google Patents
Method for preparing silk fibroin biological tissue engineering scaffold by solvent spraying Download PDFInfo
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- CN109667059B CN109667059B CN201811632028.1A CN201811632028A CN109667059B CN 109667059 B CN109667059 B CN 109667059B CN 201811632028 A CN201811632028 A CN 201811632028A CN 109667059 B CN109667059 B CN 109667059B
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
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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
- A61L27/50—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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/04—Dry spinning methods
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
- D01F4/00—Monocomponent artificial filaments or the like of proteins; Manufacture thereof
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Abstract
The technical scheme discloses a method for preparing a silk fibroin biological tissue engineering scaffold by spray, which comprises the following steps: (1) dissolving degummed silk by using a solvent to obtain a silk fibroin solution with the concentration of 12-45%; (2) and (2) taking the silk fibroin solution prepared in the step (1) as a spinning solution, carrying out solution-jet spinning by taking a high-pressure air jet flow at the temperature of 37-120 ℃ as a stretching power, and receiving the spinning by using a fiber receiver. The silk fibroin biological tissue engineering scaffold prepared by the preparation method has high porosity, good pore canal connectivity and large specific surface area, can provide a good microenvironment for the growth of cells, and is beneficial to the adhesion, differentiation and proliferation of the cells; the silk fibroin biological tissue engineering scaffold prepared by the technical method can be used for tissue engineering scaffolds such as blood vessels, urethra, cartilage, bone and the like, and can also be used for filtering blood after surface treatment.
Description
Technical Field
The technical scheme belongs to the technical field of biomedical materials, and particularly relates to a solution-spraying preparation method of a silk fibroin biological tissue engineering scaffold.
Background
The silk fibroin biological tissue engineering scaffold membrane can simulate the structure and function of extracellular matrix to a certain extent, provides ideal growth, proliferation and differentiation microenvironment for cells, and is widely applied to various fields including tissue engineering research of cartilage, bone, blood vessels, skin and the like within a few years. The electrostatic spinning technology is the only preparation technology of the silk fibroin biological tissue engineering scaffold at present, and the mechanical property of the silk fibroin biological tissue engineering scaffold can meet the mechanical strength requirements of different tissue engineering applications. However, the mechanical properties of the silk fibroin biological tissue engineering scaffold obtained by primary spinning are poor, and the mechanical strength requirement required by the application of the tissue engineering scaffold can be met only after certain post-treatment process treatment. The post-treatment process increases the production cost of the silk fibroin biological tissue engineering scaffold.
The preparation of the silk fibroin biological tissue engineering scaffold by the electrostatic spinning technology has the following disadvantages: (1) higher voltages are required, there are drawbacks in operational safety, and it is not easy to apply in industrial production on a large scale; (2) in the electrostatic spinning process with extremely high speed, the single electrostatic stretching force cannot promote the silk fibroin molecules to be converted into a beta-folding structure from a random coil/alpha-helical structure in an aqueous solution state, so that the silk fibroin is difficult to crystallize and rearrange, and the collected fibers have low crystallinity and poor mechanical property; (3) the electrostatic spinning production technology is just entering the industrial trial production stage, and a plurality of difficulties need to be overcome.
Disclosure of Invention
The technical scheme provides a solution-spraying preparation method of a silk fibroin biological tissue engineering scaffold, the preparation method can simulate the animal spinning process in nature, and the silk fibroin biological tissue engineering scaffold prepared by the preparation method has good mechanical properties.
In order to achieve the above purpose, the technical means adopted by the technical scheme is as follows.
A method for preparing a silk fibroin biological tissue engineering scaffold by solvent spraying comprises the following steps:
(1) dissolving and dialyzing the degummed silk by using a solvent to obtain a silk fibroin solution with the concentration of 12-45%;
(2) and (2) taking the silk fibroin solution prepared in the step (1) as a spinning solution, and carrying out solution-jet spinning by taking a high-pressure air jet flow at the temperature of 37-120 ℃ as a stretching power.
Further, the solvent for dissolving the degummed silk is LiBr water solution or ethanol/CaCl water solution.
Further, the method for preparing the silk fibroin biological tissue engineering scaffold by solution spraying also comprises the following steps: and (2) adding a pH value regulator into the silk fibroin solution prepared in the step (1) to regulate the pH value, wherein the added pH value regulator is metal ions or inorganic nano powder.
Further, the metal ion is one of calcium ion and silver ion.
Further, the inorganic nano powder is one of hydroxyapatite and tricalcium phosphate.
Further, when the fiber receiver adopts a fiber receiving flat plate, a silk fibroin fiber membrane is obtained.
Furthermore, when the fiber receiver adopts a fiber receiving roller, the silk fibroin continuous long fiber membrane which is stretched in a single direction and arranged in a single direction is obtained.
Further, the high-pressure air jet flow in the step (2) is dried compressed air, and the relative humidity of the high-pressure air jet flow is 10% -40%.
Furthermore, the injection pressure of the high-pressure air injection flow is 10MPa to 30 MPa.
The beneficial effects of this technical scheme do: the diameter of silk fibroin fiber in the silk fibroin biological tissue engineering scaffold prepared by the preparation method in the technical scheme is micron and submicron grade, even can reach nanometer grade, and can imitate the structure of human extracellular matrix to the maximum extent; the preparation method in the technical scheme does not need voltage, does not adopt an organic solvent, has high operability and safety, is environment-friendly, and can be applied to industrial production in a large scale; the preparation method in the technical scheme can enable the porosity in the prepared silk fibroin biological tissue engineering scaffold to meet the requirements of different cell growth by adjusting the processing parameters; the silk fibroin biological tissue engineering scaffold prepared by the preparation method has high porosity, good pore canal connectivity and large specific surface area, can provide a good microenvironment for the growth of cells, and is beneficial to the adhesion, differentiation and proliferation of the cells; the silk fibroin fibrous membrane prepared by the technical method can be used for tissue engineering scaffolds such as blood vessels, urethra, cartilage, bone and the like, and can also be used for filtering blood after surface treatment.
Detailed Description
The technical scheme of the invention is further explained by combining specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Example 1
A method for preparing a silk fibroin bioengineering bracket by solvent spraying comprises the following steps: (1) degumming silkworm cocoons by using a sodium carbonate aqueous solution to obtain degummed silk; dissolving degummed silk by using 9mol/L lithium bromide aqueous solution as a solvent, and dissolving the degummed silk for 2 hours in a water bath environment at 40 ℃ to obtain a silk fibroin aqueous solution; (2) centrifuging, filtering, dialyzing and concentrating to obtain 12 wt% silk fibroin spinning solution; (3) and (2) carrying out solution-jet spinning on the silk fibroin spinning solution by taking a high-pressure air jet flow with the temperature of 120 ℃ and the relative humidity of 10% as a stretching power, wherein the jet pressure of the high-pressure air jet flow is 15MPa, and receiving the spinning by adopting a fiber receiving flat plate to prepare the silk fibroin fiber membrane, wherein the average diameter of the fiber is 6 microns.
Example 2
A method for preparing a silk fibroin bioengineering bracket by solvent spraying comprises the following steps: (1) degumming silkworm cocoons by using a sodium carbonate aqueous solution to obtain degummed silk, dissolving the degummed silk by using a 9mol/L lithium bromide aqueous solution as a solvent, and dissolving the degummed silk for 12 hours in a water bath environment at 40 ℃ to obtain a silk fibroin aqueous solution; (2) after centrifugation, filtration, dialysis and concentration to 20 wt%, adding CaCl until the concentration of calcium ions in the silk fibroin aqueous solution is 0.3mol/L, and continuing to concentrate until the concentration of the silk fibroin is 45 wt%; (3) and (2) carrying out solution-jet spinning on the silk fibroin spinning solution by taking a high-pressure air jet flow with the temperature of 37 ℃ and the humidity of 50 wt% as a stretching power, wherein the jet pressure of the high-pressure air jet flow is 30MPa, and receiving the spinning by adopting a fiber receiving flat plate to obtain the silk fibroin fiber membrane, wherein the average diameter of the fiber is 3 mu m.
Example 3
A method for preparing a silk fibroin bioengineering bracket by solvent spraying comprises the following steps: (1) degumming silkworm cocoons by using a sodium carbonate aqueous solution to obtain degummed silk, dissolving the degummed silk by using a 7mol/L calcium chloride/ethanol aqueous solution as a solvent, and dissolving for 6 hours in a water bath environment at 40 ℃ to obtain a silk fibroin aqueous solution; (2) centrifuging, filtering, dialyzing and concentrating to obtain spinning solution with the silk fibroin concentration of 30 wt%; (3) and (2) carrying out solution-jet spinning on the silk fibroin spinning solution by taking a high-pressure air jet flow with the temperature of 100 ℃ and the humidity of 25 wt% as a stretching power, wherein the jet pressure of the high-pressure air jet flow is 10MPa, and receiving the spinning by adopting a fiber receiving flat plate to obtain the silk fibroin fiber membrane, wherein the average diameter of the fiber is 5 microns.
Example 4
A method for preparing a silk fibroin bioengineering bracket by solvent spraying comprises the following steps: (1) degumming silkworm cocoons by using a sodium carbonate aqueous solution to obtain degummed silk, dissolving the degummed silk by using a 7mol/L calcium chloride/ethanol aqueous solution as a solvent, and dissolving the degummed silk for 12 hours in a water bath environment at 40 ℃ to obtain a silk fibroin aqueous solution; (2) centrifuging, filtering, dialyzing and concentrating to obtain spinning solution with the silk fibroin concentration of 12 wt%; (3) and (2) carrying out solution-jet spinning on the silk fibroin spinning solution by taking a high-pressure air jet flow with the temperature of 100 ℃ and the humidity of 10 wt% as a stretching power, wherein the jet pressure of the high-pressure air jet flow is 25MPa, and receiving the spinning by adopting a fiber receiving flat plate to obtain the silk fibroin fiber membrane, wherein the average diameter of the fiber is 200 nm.
While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the present invention is not limited to the embodiments, but various equivalent modifications and substitutions can be made without departing from the spirit of the present invention, and the equivalents and substitutions are intended to be included in the scope of the present invention as defined by the appended claims.
Claims (6)
1. A method for preparing a silk fibroin biological tissue engineering scaffold by solvent spraying is characterized by comprising the following steps:
(1) dissolving degummed silk by using LiBr aqueous solution or ethanol/CaCl aqueous solution as a solvent to obtain silk fibroin solution with the concentration of 12-45%;
(2) taking the silk fibroin solution prepared in the step (1) as a spinning solution, carrying out spray spinning by taking a high-pressure air jet flow at the temperature of 37-120 ℃ as a stretching power, and receiving the spinning by using a fiber receiver to prepare the silk fibroin biological tissue engineering scaffold; the high-pressure air jet flow is dried compressed air, and the relative humidity of the high-pressure air jet flow is 10-50%; the injection pressure in the high-pressure air injection flow is 10MPa to 30 MPa.
2. The solution spraying preparation method of the silk fibroin biological tissue engineering scaffold as claimed in claim 1, further comprising the steps of: and (2) adding a pH value regulator into the silk fibroin solution prepared in the step (1) to regulate the pH value, wherein the added pH value regulator is metal ions or inorganic nano powder.
3. The solution spraying preparation method of the silk fibroin biological tissue engineering scaffold as claimed in claim 2, wherein the metal ion is one of calcium ion and silver ion.
4. The method for preparing a silk fibroin biological tissue engineering scaffold by spray coating according to claim 2, wherein the inorganic nanopowder is one of hydroxyapatite and tricalcium phosphate.
5. The method for preparing the silk fibroin biological tissue engineering scaffold by solvent spraying as claimed in claim 1, wherein the silk fibroin fibrous membrane is obtained when the fiber receiver adopts a fiber receiving flat plate.
6. The method for preparing the silk fibroin biological tissue engineering scaffold by solvent spraying according to claim 1, wherein when the fiber receiver adopts a fiber receiving roller, a unidirectionally stretched and unidirectionally and orderly arranged silk fibroin continuous long fiber membrane is obtained.
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| CN109529125B (en) * | 2018-12-28 | 2023-01-24 | 佛山科学技术学院 | Method for preparing biological tissue engineering scaffold by solvent spraying |
| CN110585480B (en) | 2019-10-08 | 2021-10-29 | 东南大学 | Forming method of self-supporting silk fibroin catheter stent |
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