CN115737909A - Preparation method and application of nanofiber material taking polycaprolactone as matrix - Google Patents
Preparation method and application of nanofiber material taking polycaprolactone as matrix Download PDFInfo
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Abstract
The invention provides a nanofiber material taking polycaprolactone as a matrix and a preparation method thereof, wherein the preparation method comprises the following steps: mixing and dissolving the polycaprolactone master batch and an organic solvent according to a certain proportion, preparing the obtained slurry into a fiber film through electrostatic spinning, carrying out surface modification and drug loading after drying, and drying and sterilizing to obtain the nanofiber material. The nanofiber material taking polycaprolactone as a matrix provided by the invention has relatively stable performance and chemical stability in vivo, has good biocompatibility, and is excellent in tissue repair promotion and drug loading performance.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a nanofiber material taking polycaprolactone as a matrix and a preparation method and application thereof.
Background
Biomaterials are a new class of high-tech materials used in the diagnosis, treatment, repair or replacement of human tissues, organs or to enhance their functions, which relate to the health of hundreds of millions of people and are a necessity for the health of human beings. The application of the medicine not only saves the lives of tens of thousands of critically ill patients, obviously reduces the death rate of major diseases such as cardiovascular diseases, cancers, wounds and the like, but also plays an important role in improving the life quality and health level of the patients and reducing the medical cost.
Gastrointestinal tract and pancreaticobiliary tract are important components of the digestive system, and relate to various links such as food conveying, digestion, absorption, discharge and the like, digestive tract diseases may cause the esophagus, intestinal tract, biliary tract and pancreas of patients to be narrow, and the digestive tract of some patients is blocked by malignant tumors. When these cancer patients no longer prefer surgical treatment and are intolerant to chemotherapy or radiation therapy, stent implantation has been used as an excellent palliative treatment modality because it allows immediate relief from symptoms in the patient. In addition, temporary internal stenting procedures may be beneficial in treating these several digestive tract disorders, and may help ensure that bile, pancreatic juice, or fluid accumulation flows into the gastrointestinal tract after endoscopic or surgical procedures. Stent-graft therapy is of great advantage and appeal for the treatment of strictures of the digestive tract or for pain relief and for promoting tissue repair.
Endogenous electric fields and electric stimulation are widely proved to have a promoting effect on cell growth and wound healing, conducting polymers such as polyaniline, polypyrrole and polythiophene have no cytotoxicity and can be used for tissue engineering, and after the surface of the material is modified and added with the conducting additives, the purpose of promoting tissue repair can be achieved by utilizing the endogenous electric fields or external electric stimulation. Heparan sulfate proteoglycan has wide action in mammals, and has important action on maintaining the homeostasis of the human body system. They have a wide range of regulatory effects on the digestive, respiratory, endocrine, nervous, urinary, immune and circulatory systems of the human body. Polydopamine with excellent biocompatibility has been widely used for surface modification of biomaterials. The reason for this is that the groups of polydopamine are mainly phenolic hydroxyl, amine and imine groups, which can further improve the surface properties of biomaterials by covalently bonding target molecules. Under the bonding effect of dopamine, the prepared nanofiber material can play a role in transporting and carrying the conductive auxiliary and the heparan sulfate proteoglycan.
Polycaprolactone is a hydrophobic semi-crystalline polymer whose crystallinity tends to decrease as molecular weight increases. The good solubility, low melting point and excellent blending compatibility of polycaprolactone have prompted extensive research into its potential applications in the biomedical field. Polycaprolactone has gained popularity in the early seventies, but degradation cycles of polycaprolactone up to 1 year or more have been rapidly replaced by polyglycolide, polylactide, and copolymers thereof having shorter degradation cycles. The advent of tissue engineering, which began to resurge interest in polycaprolactone almost 20 years ago, recognized that polycaprolactone has superior rheological and viscoelastic properties over many absorbable polymer counterparts, making it easy to manufacture and manipulate into a wide range of scaffolds. In addition, the fact that many drug delivery devices manufactured using polycaprolactone have been FDA approved and CE labeled provides a faster avenue for the market.
In the field of biological materials, polycaprolactone has good biocompatibility, excellent rheological and viscoelastic properties, relatively stable chemical properties and simple processing mode, and can be designed and prepared according to actual application. Polycaprolactone is dissolved in an organic solvent, nanofibers can be obtained through electrostatic spinning, the nanofiber has a large specific surface area, a surface conducting layer is obtained through in-situ oxidative polymerization, the surface conducting layer is soaked in heparan sulfate proteoglycan solution to load the drug, and the functions of transportation and loading are achieved through dopamine. The polycaprolactone nanofiber material after surface modification and drug loading can improve the adhesion of nutrients such as protein and the like on the surface of the material, increase biocompatibility, reduce inflammation and cytotoxicity, and has excellent performance in the aspects of promoting cell growth and tissue repair.
Disclosure of Invention
The invention provides a nano-fiber material taking polycaprolactone as a matrix and a preparation method thereof, aiming at improving the functionality of the polycaprolactone and the stability of the polycaprolactone in different acid-base in-vivo environments of pancreatic bile duct, gastrointestinal tract and the like, and the polycaprolactone nano-fiber material after surface modification and drug loading treatment can improve the adhesion of nutrients such as protein and the like on the surface of the material, increase the biocompatibility, reduce the inflammation and cytotoxicity and has excellent performance in promoting cell growth and tissue repair
In order to achieve the above purpose, the utility model adopts the following scheme:
a preparation method of a nanofiber material taking polycaprolactone as a matrix is characterized by comprising the following steps:
s1, mixing and dissolving a polycaprolactone master batch and an organic solvent to obtain slurry;
s2, preparing the obtained slurry into a polycaprolactone fiber film through electrostatic spinning;
and S3, drying the polycaprolactone fiber film, carrying out surface modification and drug loading, and drying and sterilizing to obtain the nanofiber material.
Further, the nanofiber material is composed of a polycaprolactone fiber film, a modified material and a loaded drug.
Further, the polycaprolactone fiber film is prepared by dissolving polycaprolactone master batch in an organic solvent according to the mass fraction (10-30%), then carrying out electrostatic spinning, and drying.
Further, the organic solvent mainly comprises two solvents A and B according to the mass ratio of 7, wherein A is one of dichloromethane and dichloroethane, and B is one of N, N-dimethylformamide and N, N-dimethylacetamide.
Further, the surface modification is to soak the prepared polycaprolactone fiber film in dopamine solution at 15-45 ℃ for 10-48 h, and to place the film in inorganic acid solution containing oxidant and conductive polymer at-15-5 ℃ for 10-48 h after the soaking is finished.
Furthermore, the dopamine solution has a solute of dopamine, a solvent of Tris-HCl buffer solution or phosphate buffer solution, and the concentration of the dopamine is 0.01-100 mg/mL.
Further, the inorganic acid solution containing an oxidant and a conductive polymer, wherein the oxidant is one of ammonium persulfate and ferric chloride, the conductive polymer is one of aniline, pyrrole and thiophene, the inorganic acid is one of hydrochloric acid, sulfuric acid and perchloric acid, and the molar ratio of the oxidant: conductive polymer: inorganic acid = (0.005 to 0.1): (0.005-0.1): (0.1-10) adding deionized water to a constant volume of 1L to prepare the inorganic acid solution.
Further, the drug loading is mainly carried out by a soaking method, the used drug is heparan sulfate proteoglycan, ultrapure water is used as a solvent, the drug content is 1-100 mg/ml, and the fiber film after the surface modification is soaked in the heparan sulfate proteoglycan aqueous solution for 10-48 hours at the temperature of 30-40 ℃.
Further, the sterilization is any one of ultraviolet sterilization, dry heat sterilization and high-pressure steam sterilization.
Furthermore, the preparation method of the nanofiber material with polycaprolactone as the matrix can be used as a culture and proliferation carrier of cells such as vascular endothelial cells (HUVEC) gastric parietal cells, gastric mucosal cells, islet cells and the like in vitro, and can also be prepared into a stent for repairing tissues related to digestive systems such as gastrointestinal tracts, pancreatic ducts and the like.
In summary, compared with the prior art, the invention has the beneficial effects that:
the nanofiber material taking polycaprolactone as a matrix provided by the invention has relatively stable performance and chemical stability in vivo, has good biocompatibility, and is excellent in tissue repair promotion and drug loading performance.
Drawings
FIG. 1 is a flow chart of the preparation of polycaprolactone nanofiber.
Fig. 2 is a graph comparing the control group and the experimental group.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every intervening value, to the extent any stated or intervening value in a stated range, and every other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the documents are cited. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The description and examples are intended to be illustrative only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
In the present invention, all the raw materials are conventional commercially available products.
Example 1
3g of the polycaprolactone plastic master batch was added to a mixed solution containing 18.9g of dichloromethane, 8.1gN, N-dimethylformamide and stirred at 50 ℃ for 4 hours. The obtained slurry was put into a 5ml syringe, and the spinning voltage was set at 20KV, the reception distance was 15cm, and the injection speed was 0.8ml/h. After the electrostatic spinning is finished, the obtained product is placed into a vacuum drying oven and dried for 24 hours at the temperature of 40 ℃.
Cutting the dried polycaprolactone nanofiber membrane into a wafer with the thickness of 10mm by using a cutter, immersing the wafer into 2mg/ml dopamine solution for 24 hours after ultrasonic cleaning, taking out the wafer, washing the wafer for three times by using deionized water, and drying the wafer. The dried wafer was immersed in 100ml of a mineral acid solution for 24h, the solution having the composition and molar ratio: ammonium persulfate: aniline: hydrochloric acid = (0.01 mol) (0.01 mol) (1 mol) and deionized water are added to the solution to make a solution with a constant volume of 1L. The surface-modified fiber film was immersed in 10mg/ml aqueous heparan sulfate proteoglycan solution at 35 ℃ for 24 hours. Drying, and performing ultraviolet sterilization for 30min to obtain the nanofiber material taking polycaprolactone as a matrix.
Putting the polycaprolactone wafer into a forty-eight orifice plate, putting one prepared polycaprolactone wafer into each orifice, then inoculating 8000 cells into each orifice, putting the cells into an incubator to be cultured for 1, 3 and 7 days, and taking a blank polycaprolactone wafer as a control. The cell activity was evaluated by the MTT method.
The cell activities of the control groups at 1, 3 and 7 days are as follows: 0.30, 0.41, 0.45.
The cell activities in experimental groups 1, 3 and 7 days were: 0.70, 0.82, 0.93.
Compared with a control group, the cell activity of an experimental group is greatly improved, and the biocompatibility is effectively improved.
Example 2
3g of the polycaprolactone plastic master batch was added to a mixed solution containing 11.9g of dichloromethane and 5.1gN, N-dimethylformamide and stirred at 45 ℃ for 5h. The obtained slurry was charged into a 5ml syringe, and the spinning voltage was set at 20KV, the reception distance was 15cm, and the injection speed was 0.8ml/h. After the electrostatic spinning is finished, the obtained product is placed into a vacuum drying oven and dried for 20 hours at the temperature of 45 ℃.
And cutting the dried polycaprolactone nanofiber membrane into a wafer with the thickness of 10mm by using a cutter, immersing the wafer in a dopamine solution with the concentration of 4mg/ml for 18 hours after ultrasonic cleaning, taking out the wafer, washing the wafer with deionized water for three times, and drying the wafer. Immersing the dried wafer in 100ml of inorganic acid solution for 12 hours, wherein the solution comprises the following components in molar ratio: ammonium persulfate: pyrrole: perchloric acid = (0.05 mol) (0.05 mol) (5 mol) and deionized water are added to make a solution with a constant volume of 1L. The surface-modified fiber film was immersed in 20mg/ml aqueous heparan sulfate proteoglycan solution at 40 ℃ for 18 hours. Drying, and performing ultraviolet sterilization for 30min to obtain the nanofiber material taking polycaprolactone as a matrix.
Putting the polycaprolactone wafer into a forty-eight orifice plate, putting one prepared polycaprolactone wafer into each orifice, then adding 50 mu L of matrigel into each orifice, and then putting the orifice plate into a 37 ℃ incubator for 4h to solidify the matrigel into gel. Then, 8000 cells are inoculated in each hole, the cells are vascular endothelial cells, the cells are placed in an incubator to be cultured for 18h, calcein (AM) is used for staining, an inverted fluorescence microscope is used for observing vascularization conditions, and a blank polycaprolactone wafer is used as a control.
Compared with a control component, the activity of the blood vessel of the experimental group is greatly improved, and the tissue repair is promoted.
Example 3
6g of polycaprolactone plastic master batch is taken and added into a mixed solution containing 9.8g of dichloroethane, 4.2gN and N-dimethylacetamide, and the mixture is uniformly stirred for 4 hours at the temperature of 45 ℃. The obtained slurry was charged into a 5ml syringe, and the spinning voltage was set at 20KV, the reception distance was 15cm, and the injection speed was 0.8ml/h. After the electrostatic spinning is finished, the obtained product is placed into a vacuum drying oven and dried for 24 hours at the temperature of 40 ℃.
Cutting the dried polycaprolactone nanofiber membrane into a wafer with the thickness of 10mm by using a cutter, immersing the wafer into 3mg/ml dopamine solution for 24 hours after ultrasonic cleaning, taking out the wafer, washing the wafer for three times by using deionized water, and drying the wafer. The dried wafer was immersed in 100ml of a mineral acid solution for 12h, the solution having the composition and molar ratio: iron chloride: thiophene: sulfuric acid = (0.02 mol) (0.02 mol) (2 mol) deionized water is added to the solution to make a solution with a constant volume of 1L. The fiber film after the surface modification is soaked in 50mg/ml heparan sulfate proteoglycan water solution for 12 hours at the temperature of 30 ℃. Drying, and performing ultraviolet sterilization for 30min to obtain the nanofiber material taking polycaprolactone as a matrix.
The degradability of the prepared polycaprolactone nanofiber wafer under different pH values is measured by adopting a mass loss method, the mass of the prepared polycaprolactone nanofiber wafer before an experiment is recorded after drying is finished, the wafer is respectively soaked in solutions with the pH =8.5, the pH =7.4 and the pH =1, incubation is carried out for 21 days at 37 ℃, the solutions are replaced once every five days, and the polycaprolactone nanofiber wafer is weighed after being dried for a constant weight at 40 ℃. And a blank polycaprolactone disc was used as a control.
Mass loss of control group: 20% (PH = 8.5), 15% (PH = 7.4), 14% (PH = 1).
Mass loss in experimental groups: 11% (PH = 8.5), 14% (PH = 7.4), 8% (PH = 1).
Compared with a control group, the stability of the experimental group is improved under a partial acid or partial alkali environment.
Claims (10)
1. A preparation method of a nanofiber material taking polycaprolactone as a matrix is characterized by comprising the following steps:
s1, mixing and dissolving a polycaprolactone master batch and an organic solvent to obtain slurry;
s2, preparing the obtained slurry into a polycaprolactone fiber film through electrostatic spinning;
and S3, drying the polycaprolactone fiber film, carrying out surface modification and drug loading, and drying and sterilizing to obtain the nanofiber material.
2. The preparation method of the nanofiber material taking polycaprolactone as the matrix according to claim 1, wherein the nanofiber material is composed of a polycaprolactone fiber film, a modified material and a loaded drug.
3. The preparation method of the nanofiber material taking polycaprolactone as the matrix according to claim 1, wherein the polycaprolactone fiber film is prepared by dissolving polycaprolactone master batch in an organic solvent according to the mass fraction (10% -30%), performing electrostatic spinning, and drying.
4. The method for preparing a nanofiber material taking polycaprolactone as a matrix according to claim 3, wherein the organic solvent mainly comprises two solvents A and B according to a mass ratio of 7.
5. The method for preparing the nanofiber material taking polycaprolactone as the matrix according to claim 1, wherein the surface modification is carried out by soaking the prepared polycaprolactone fiber film in dopamine solution at 15-45 ℃ for 10-48 h, and after the soaking, placing the polycaprolactone fiber film in inorganic acid solution containing an oxidant and a conductive polymer at-15-5 ℃ for 10-48 h.
6. The method for preparing a nanofiber material taking polycaprolactone as a matrix according to claim 5, wherein the dopamine solution is prepared by using dopamine as a solute and Tris-HCl buffer or phosphate buffer as a solvent, and the concentration of the dopamine is 0.01-100 mg/mL.
7. The method for preparing the nanofiber material taking polycaprolactone as the matrix according to claim 6, wherein the inorganic acid solution containing an oxidant and a conductive polymer is one of ammonium persulfate and ferric chloride, the conductive polymer is one of aniline, pyrrole and thiophene, the inorganic acid is one of hydrochloric acid, sulfuric acid and perchloric acid, and the molar ratio of the oxidant: conductive polymer: inorganic acid = (0.005 to 0.1): (0.005-0.1): (0.1-10) adding deionized water to a constant volume of 1L to prepare the inorganic acid solution.
8. The method for preparing a nanofiber material taking polycaprolactone as a matrix according to claim 1, wherein the drug loading mainly adopts a soaking method, the used drug is acetamide sulfate heparin proteoglycan, ultrapure water is used as a solvent, the drug content is 1-100 mg/ml, and the fiber film after the surface modification is soaked in the acetamide sulfate heparin proteoglycan aqueous solution for 10-48 h at 30-40 ℃.
9. A nanofiber material taking polycaprolactone as a matrix is characterized in that: the production method according to any one of claims 1 to 8.
10. Use of a nanofiber material based on polycaprolactone according to claim 9, characterized in that: the nanofiber material can be used as a culture proliferation carrier of cells in vitro, such as vascular endothelial cells (HUVEC), gastric parietal cells, gastric mucosal cells and pancreatic islets, and can also be prepared into a stent for repairing tissues related to a digestive system, such as gastrointestinal tracts, pancreas and gall bladder.
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| CN108193501A (en) * | 2018-01-24 | 2018-06-22 | 哈尔滨工业大学 | A kind of conductive shapes memory membrane, preparation method and its electric drive method |
| CN114000262A (en) * | 2021-12-06 | 2022-02-01 | 广东工业大学 | Drug-loaded polydopamine-coated nanofiber dressing and preparation method thereof |
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| CN108193501A (en) * | 2018-01-24 | 2018-06-22 | 哈尔滨工业大学 | A kind of conductive shapes memory membrane, preparation method and its electric drive method |
| CN114000262A (en) * | 2021-12-06 | 2022-02-01 | 广东工业大学 | Drug-loaded polydopamine-coated nanofiber dressing and preparation method thereof |
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