Beta-1, 3 / alpha-1, 3-glucan nanofiber as well as preparation method and application thereof

Abstract

The invention relates to a beta-1, 3 / alpha-1, 3-glucan nanofiber and a preparation method and application thereof.The preparation method comprises the steps that 5.5 wt%-6.5 wt% of a drug carrier and 10 wt%-20 wt% of beta-1, 3 / alpha-1, 3-glucan are added into a pre-prepared solution system at the same time and stirred and mixed, and an electrostatic spinning solution is obtained; and performing electrostatic spinning processing on the electrostatic spinning solution by adopting an electrostatic spinning process to obtain the beta-1, 3 / alpha-1, 3-glucan nanofiber. The beta-1, 3 / alpha-1, 3-glucan nanofiber prepared by the method has better water solubility, good dispersity and better anti-inflammatory and antibacterial curative effects; meanwhile, polycaprolactone which is good in biocompatibility, free of immunogenicity, biodegradable and high in mechanical strength is selected as a base material of the nanofiber, so that the beta-1, 3 / alpha-1, 3-glucan nanofiber is high in mechanical performance and good in compatibility; and by controlling the parameters of the electrostatic spinning process, the diameter and the form of the nanofiber can be accurately controlled, no particle crosstalk state exists, and different application requirements can be met.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to a beta-1,3 / alpha-1,3-glucan nanofiber, a preparation method and application thereof. Background Art

[0002] β-1,3 / α-1,3-glucan is a bioactive polysaccharide widely found in the cell walls of natural plants and microorganisms such as yeast, mushrooms, oats, and rice bran. It can improve skin immunity, accelerate wound healing, and enhance the body's resistance to bacterial, fungal and viral infections by stimulating the body's nonspecific immune response. Therefore, it has been used in clinical treatments such as wound care management and metabolic dysbiosis.

[0003] Traditional wound dressings, such as dry gauze, hydrocolloid dressings, transparent film dressings, hydrogel dressings, etc., have problems such as bacterial invasion, poor absorption of exudate, poor mechanical properties, and obvious scars after treatment. At the same time, they have low drug loading, uneven drug distribution, and poor drug release, which affects the efficacy of the drug. β-1,3 / α-1,3-glucan nanofibers are more gentle and have fewer side effects. As a treatment for skin wound healing, they have better efficacy.

[0004] The traditional preparation method of glucan nanofibers has some shortcomings, such as uneven fiber diameter, low production efficiency, poor product solubility, low bioavailability, etc., which leads to insufficient efficacy. In addition, the traditional β-1,3 / α-1,3-glucan fibers have insufficient physical and mechanical protection, limited gas exchange capacity, uneven distribution of active ingredients, low wound healing potential, low biological activity and affinity, and difficulty in controlling drug release. Therefore, it is necessary to improve the preparation method of β-1,3 / α-1,3-glucan nanofibers to obtain β-1,3 / α-1,3-glucan nanofibers with better performance. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a β-1,3 / α-1,3-glucan nanofiber, a preparation method and application thereof, which has the advantages of a simple preparation method and better performance of the obtained nanofiber.

[0006] To achieve the above objectives and other related objectives, the present invention provides the following technical solutions: The first aspect of the disclosed embodiment of the present invention provides a method for preparing β-1,3 / α-1,3-glucan nanofibers, comprising: Adding 5.5-6.5 wt % of the drug carrier and 10-20 wt % of β-1,3 / α-1,3-glucan to the pre-formulated solution system and stirring and mixing to obtain an electrospinning solution; The electrospinning solution was electrospun using an electrospinning process to obtain β-1,3 / α-1,3-glucan nanofibers.

[0007] As a preferred embodiment of the invention, the solution system is a formic acid / acetic acid solution with a ratio of (65-70) / (30-35).

[0008] As a preferred embodiment of the invention, the drug carrier is polycaprolactone.

[0009] As a preferred embodiment of the invention, the parameters of the electrospinning process are: the distance from the needle to the collector is 11~13cm, the electrospinning voltage is 25~35 kV, the electrospinning room temperature is 23~27℃, the relative humidity is 55~65%, and the flow rate is set to 1.7~2.8mL / h.

[0010] According to a second aspect of the disclosed embodiments of the present invention, there is provided a β-1,3 / α-1,3-glucan nanofiber, which is prepared by the preparation method described in the first aspect.

[0011] The third aspect of the disclosed embodiments of the present invention provides the use of β-1,3 / α-1,3-glucan nanofibers prepared by the preparation method described in the first aspect in wound care drugs.

[0012] As a preferred embodiment of the invention, the wound care medicine comprises a dressing.

[0013] As described above, the present invention has the following beneficial effects: The embodiment of the present invention provides a β-1,3 / α-1,3-glucan nanofiber, a preparation method and application thereof, wherein the preparation method comprises: adding 5.5-6.5wt% of a drug carrier and 10-20wt% of β-1,3 / α-1,3-glucan to a pre-formulated solution system and stirring and mixing to obtain an electrospinning solution; and electrospinning the electrospinning solution using an electrospinning process to obtain the β-1,3 / α-1,3-glucan nanofiber. The β-1,3 / α-1,3-glucan nanofibers prepared by this method have good water solubility and dispersibility, and have good anti-inflammatory and antibacterial effects; at the same time, polycaprolactone with good biocompatibility, non-immunogenicity, biodegradability and high mechanical strength is selected as the substrate of the nanofibers, so that the β-1,3 / α-1,3-glucan nanofibers have high mechanical properties and good compatibility; and by controlling the parameters of the electrospinning process, the diameter and morphology of the nanofibers can be precisely controlled, and there is no particle crosstalk state, which can meet different application requirements; and by adjusting the concentration of β-1,3 / α-1,3-glucan, the hydrophilicity of the electrospinning solution can be changed, thereby adjusting the water absorption of the β-1,3 / α-1,3-glucan nanofibers, and the surface of the nanofibers produced by the electrospinning process is smooth, and the entanglement between fibers is reduced, allowing air and water molecules to pass more easily, effectively improving the air permeability and moisture permeability. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Shown is a scanning electron microscope image of the electrospinning solution in Comparative Example 1 of the present invention.

[0015] Figure 2 Shown is the contact angle diagram of the electrospinning solution at 0s in Comparative Example 1 of the present invention.

[0016] Figure 3 Shown is the contact angle diagram of the electrospinning solution at 60s in Comparative Example 1 of the present invention.

[0017] Figure 4 Shown is a scanning electron microscope image of the electrospinning solution in Comparative Example 2 of the present invention.

[0018] Figure 5 Shown is the contact angle diagram of the electrospinning solution at 0s in Comparative Example 2 of the present invention.

[0019] Figure 6 It shows the contact angle diagram of the electrospinning solution at 60s in Comparative Example 2 of the present invention.

[0020] Figure 7 Shown is a scanning electron microscope image of the electrospinning solution in Example 1 of the present invention.

[0021] Figure 8Shown is the contact angle diagram of the electrospinning solution at 0s in Example 1 of the present invention.

[0022] Fig. 9 Shown is the contact angle diagram of the electrospinning solution at 60s in Example 1 of the present invention.

[0023] Fig.10 Shown is a scanning electron microscope image of the electrospinning solution in Example 2 of the present invention.

[0024] Fig.11 Shown is the contact angle diagram of the electrospinning solution at 0s in Example 2 of the present invention.

[0025] Fig.12 Shown is the contact angle diagram of the electrospinning solution at 60s in Example 2 of the present invention. DETAILED DESCRIPTION

[0026] The following is a description of the implementation of the present invention by means of specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0027] A first aspect of an embodiment of the present invention provides a method for preparing β-1,3 / α-1,3-glucan nanofibers, comprising: S100, adding 5.5-6.5 wt % of the drug carrier and 10-20 wt % of β-1,3 / α-1,3-glucan to the pre-formulated solution system and stirring and mixing overnight to obtain an electrospinning solution.

[0028] The solution system is a formic acid / acetic acid solution with a ratio of (65-70) / (30-35), preferably, the solution system is a formic acid / acetic acid solution with a ratio of 70 / 30, and the drug carrier is polycaprolactone, preferably, the concentration of polycaprolactone is 6wt%.

[0029] β-1,3 / α-1,3-glucan is a polysaccharide made from sucrose as raw material through fermentation with Rhizobiumpusense, alcohol precipitation, filtration, separation, drying, crushing and other processes. It is composed of glucose molecules alternately connected by β-1,3 and α-1,3 glycosidic bonds, with a clear structure and molecular formula: (C6H10O5)n, n≥615, purity ≥90. This structure gives it unique biological activity and physical properties. The β-1,3 / α-1,3-glucan used in the present invention comes from: Sichuan Hetai Xinguang Biotechnology Co., Ltd. uses patented strains of marine microorganisms, takes sucrose as the main raw material, and is obtained through biological transformation. It has a brand-new structure of β-1,3 / α-1,3-β-1,3 / α-1,3-glucan, and solves the problems of water insolubility, low purity and low activity of traditional β-1,3 / α-1,3-glucan. Preferably, the concentration of β-1,3 / α-1,3-glucan is 20 wt %.

[0030] S200, electrospinning the electrospinning solution using an electrospinning process to obtain β-1,3 / α-1,3-glucan nanofibers.

[0031] Electrospinning technology is a method of preparing ultrafine fibers by utilizing electrostatic field forces. As a new method for preparing nanofibers, it has the advantages of simple processing technology and low spinning cost. The specific method of electrospinning is as follows: adding high voltage static electricity to the electrospinning solution, the charged polymer droplets are accelerated at the top of the Taylor cone of the capillary under the action of the electric field force. When the electric field force is large enough, the polymer droplets overcome the surface tension to form a jet stream. The solvent evaporates or solidifies during the jetting process, and finally falls on the receiving device to form a fiber felt similar to a non-woven fabric. The device for electrospinning processing can adopt the existing structure.

[0032] In this embodiment, the electrospinning process is used to electrospin the electrospinning solution to obtain β-1,3 / α-1,3-glucan nanofibers: Take a 5mL syringe and fill it with 4mL of electrospinning solution. The front end of the syringe is a needle. A 23G needle is used. The distance from the needle to the collector is controlled to be 11~13cm, preferably 12cm. The stable electrospinning voltage is 25~35 kV. The specific voltage value depends on the stability of the Taylor cone. The electrospinning room temperature is controlled to be 23~27℃, the relative humidity is 55~65%, and the flow rate is set to 1.7~2.8mL / h.

[0033] The β-1,3 / α-1,3-glucan nanofibers prepared by this method have good water solubility and dispersibility, and have good anti-inflammatory and antibacterial effects; at the same time, polycaprolactone with good biocompatibility, non-immunogenicity, biodegradability and high mechanical strength is selected as the substrate of the nanofibers, so that the β-1,3 / α-1,3-glucan nanofibers have high mechanical properties and good compatibility; and by controlling the parameters of the electrospinning process, the diameter and morphology of the nanofibers can be precisely controlled, and there is no particle crosstalk state, which can meet different application requirements; and by adjusting the concentration of β-1,3 / α-1,3-glucan, the hydrophilicity of the electrospinning solution can be changed, thereby adjusting the water absorption of the β-1,3 / α-1,3-glucan nanofibers, and the surface of the nanofibers produced by the electrospinning process is smooth, and the entanglement between fibers is reduced, allowing air and water molecules to pass more easily, effectively improving the air permeability and moisture permeability.

[0034] The second aspect of the disclosed embodiment of the present invention provides a β-1,3 / α-1,3-glucan nanofiber, which is prepared by the preparation method described in the first aspect. As an emerging drug delivery system, the nanofiber can effectively deliver drugs through the skin, increase the distribution of drugs in wounds and increase the absorption of wound exudate.

[0035] The third aspect of the disclosed embodiment of the present invention provides the use of β-1,3 / α-1,3-glucan nanofibers prepared by the preparation method as described in the first aspect in wound care drugs, wherein the wound care drugs include dressings, such as fiber mats, wherein the β-1,3 / α-1,3-glucan nanofibers are very similar to natural extracellular matrix, and after forming fiber mats, the β-1,3 / α-1,3-glucan is effectively loaded, and better physical and mechanical wound protection is provided, and the uniformity and drug loading of the drug can be improved, thereby ensuring the biological activity and efficient treatment of the drug, and utilizing the advantages of the high specific surface area of ​​the nanofibers and the ability to form fiber mats to improve the biological activity of β-1,3 / α-1,3-glucan, better promote the phenotypic transformation of macrophages, exert anti-inflammatory and antioxidant effects, and better promote wound healing.

[0036] The present invention determines the hydrophilicity of the nanofibers by the contact angle of the electrospinning solution, which is described below with reference to specific examples: Comparative Example 1 A method for preparing β-1,3 / α-1,3-glucan nanofibers, comprising: S100, adding 6wt% polycaprolactone to the pre-prepared solution system and stirring and mixing overnight to obtain an electrospinning solution, wherein the solution system is a formic acid / acetic acid solution with a ratio of 70 / 30, and the scanning electron microscope image and contact angle image of the electrospinning solution are as follows: Figure 1-3 shown.

[0037] S200, electrospinning the electrospinning solution to obtain β-1,3 / α-1,3-glucan nanofibers by an electrospinning process, which is specifically as follows: a 5mL syringe is filled with 4mL of electrospinning solution, the front end of the syringe is a needle, a 23G needle is used, the distance from the needle to the collector is controlled to be 11~13cm, preferably 12cm, the stable electrospinning voltage is 25~35 kV, the specific voltage value depends on the stability of the Taylor cone, and the electrospinning room temperature is controlled to be 23~27℃, the relative humidity is 55~65%, and the flow rate is set to 1.7~2.8mL / h.

[0038] Comparative Example 2 A method for preparing β-1,3 / α-1,3-glucan nanofibers, comprising: S100, 6wt% polycaprolactone and 5wt% β-1,3 / α-1,3-glucan are added to the pre-prepared solution system and stirred overnight to obtain an electrospinning solution, wherein the solution system is a formic acid / acetic acid solution with a ratio of 70 / 30. The scanning electron microscope image and contact angle image of the electrospinning solution are as follows: Figure 4-6 shown.

[0039] S200, electrospinning the electrospinning solution to obtain β-1,3 / α-1,3-glucan nanofibers by an electrospinning process, which is specifically as follows: a 5mL syringe is filled with 4mL of electrospinning solution, the front end of the syringe is a needle, a 23G needle is used, the distance from the needle to the collector is controlled to be 11~13cm, preferably 12cm, the stable electrospinning voltage is 25~35 kV, the specific voltage value depends on the stability of the Taylor cone, and the electrospinning room temperature is controlled to be 23~27℃, the relative humidity is 55~65%, and the flow rate is set to 1.7~2.8mL / h.

[0040] Embodiment 1 A method for preparing β-1,3 / α-1,3-glucan nanofibers, comprising: S100, 6wt% polycaprolactone and 10wt% β-1,3 / α-1,3-glucan are added to the pre-prepared solution system and stirred overnight to obtain an electrospinning solution, wherein the solution system is a formic acid / acetic acid solution with a ratio of 70 / 30. The scanning electron microscope image and contact angle image of the electrospinning solution are as follows: Figure 7-9 shown.

[0041] S200, electrospinning the electrospinning solution to obtain β-1,3 / α-1,3-glucan nanofibers by an electrospinning process, which is specifically as follows: a 5mL syringe is filled with 4mL of electrospinning solution, the front end of the syringe is a needle, a 23G needle is used, the distance from the needle to the collector is controlled to be 11~13cm, preferably 12cm, the stable electrospinning voltage is 25~35 kV, the specific voltage value depends on the stability of the Taylor cone, and the electrospinning room temperature is controlled to be 23~27℃, the relative humidity is 55~65%, and the flow rate is set to 1.7~2.8mL / h.

[0042] Embodiment 2 A method for preparing β-1,3 / α-1,3-glucan nanofibers, comprising: S100, 6wt% polycaprolactone and 20wt% β-1,3 / α-1,3-glucan are added to the pre-prepared solution system and stirred overnight to obtain an electrospinning solution, wherein the solution system is a formic acid / acetic acid solution with a ratio of 70 / 30. The scanning electron microscope image and contact angle image of the electrospinning solution are as follows: Figure 10-12 shown.

[0043] S200, electrospinning the electrospinning solution to obtain β-1,3 / α-1,3-glucan nanofibers by an electrospinning process, which is specifically as follows: a 5mL syringe is filled with 4mL of electrospinning solution, the front end of the syringe is a needle, a 23G needle is used, the distance from the needle to the collector is controlled to be 11~13cm, preferably 12cm, the stable electrospinning voltage is 25~35 kV, the specific voltage value depends on the stability of the Taylor cone, and the electrospinning room temperature is controlled to be 23~27℃, the relative humidity is 55~65%, and the flow rate is set to 1.7~2.8mL / h.

[0044] By measuring the contact angles of electrospinning solutions with β-1,3 / α-1,3-glucan concentrations of 0 wt%, 5 wt%, 10 wt% and 20 wt%, it was found that with the increase of β-1,3 / α-1,3-glucan concentration, the contact angle of the electrospinning solution continued to decrease and its hydrophilicity increased. When the concentration of β-1,3 / α-1,3-glucan reaches 20 wt%, its contact angle is 69.2°. The prepared nanofibers have a certain water absorption capacity, which is beneficial to the absorption of wound exudate and improves the antibacterial properties of nanofibers. At the same time, the stability of the contact angle will affect the diameter uniformity and morphology of the nanofibers. Among the four electrospinning solutions, when the concentration of β-1,3 / α-1,3-glucan is 5 w%, the contact angle change is larger than that of other groups. The diameter uniformity and morphology of the prepared nanofibers are poor. When the concentration of β-1,3 / α-1,3-glucan is 20 wt%, the change is 4.8°, which is lower than that of β-1,3 / α-1,3-glucan at a concentration of 5 wt%.

[0045] The nanofibers prepared by the above method have a smooth surface, uniform diameter, and good morphological characteristics. By preparing different β-1,3 / α-1,3-glucan concentrations and changing the contact angle and stability of the spinning solution, nanofibers with excellent performance can be prepared, thereby improving the therapeutic effect. Compared with the prior art, β-1,3-glucan is dissolved in formic acid to prepare an electrospinning solution, and electrospinning is performed to obtain electrospun fibers with a diameter of 0.05 to 1 μm. In order to improve the spinnability of the spinning solution, pharmaceutical-grade synthetic polymers are usually added thereto. The prepared nanofibers are too fragile, and a certain number of nanofibers are bent. Many cracks perpendicular to the fiber axis are obvious, and they cannot be used as reinforcing materials for composite materials. There are common problems such as uneven diameter, rough surface, low drug loading, low yield, and poor mechanical properties of the nanofibers. The preparation process is complicated, involving ultrasonic grinding, homemade equipment, etc., and traditional β-1,3 / α-1,3-glucan fibers have insufficient physical and mechanical protection, limited gas exchange capacity, uneven distribution of active ingredients, low wound healing potential, low biological activity and affinity, and difficulty in drug release control.

[0046] The present invention selects polycaprolactone with good biocompatibility, non-immunogenicity, biodegradability and high mechanical strength as the substrate of the nanofiber, so that the β-1,3 / α-1,3-glucan nanofiber has high mechanical properties and good compatibility, and loads β-1,3 / α-1,3-glucan to prepare the nanofiber through electrospinning technology. The prepared β-1,3 / α-1,3-glucan nanofiber is very similar to the natural extracellular matrix and forms a fiber mat, which provides better physical and mechanical wound protection, while improving the uniformity and drug loading of the drug, ensuring the biological activity and efficient treatment of the drug, and utilizing the advantages of the high specific surface area of ​​nanofibers and the ability to form a fiber mat to improve the biological activity of β-1,3 / α-1,3-glucan, better promote the phenotypic transformation of macrophages, exert anti-inflammatory and antioxidant effects, and better promote wound healing.

[0047] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A method for preparing β-1,3 / α-1,3-glucan nanofibers, characterized in that: include: Adding 5.5-6.5 wt % of the drug carrier and 10-20 wt % of β-1,3 / α-1,3-glucan to the pre-formulated solution system and stirring and mixing to obtain an electrospinning solution; The electrospinning solution was electrospun using an electrospinning process to obtain β-1,3 / α-1,3-glucan nanofibers.

2. The method for preparing β-1,3 / α-1,3-glucan nanofibers according to claim 1, characterized in that: The solution system is a formic acid / acetic acid solution with a ratio of (65-70) / (30-35).

3. The method for preparing β-1,3 / α-1,3-glucan nanofibers according to claim 1 or 2, characterized in that: The drug carrier is polycaprolactone.

4. The method for preparing β-1,3 / α-1,3-glucan nanofibers according to claim 1, characterized in that: The parameters of the electrospinning process are: the distance from the needle to the collector is 11-13 cm, the electrospinning voltage is 25-35 kV, the electrospinning room temperature is 23-27° C., the relative humidity is 55-65%, and the flow rate is set to 1.7-2.8 mL / h.

5. A β-1,3 / α-1,3-glucan nanofiber, characterized in that: Prepared by the preparation method according to any one of claims 1 to 4.

6. Use of the β-1,3 / α-1,3-glucan nanofibers prepared by the preparation method according to any one of claims 1 to 4 in wound care drugs.

7. The use according to claim 6, characterized in that: The wound care medicament includes a dressing.

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