Antibacterial polyvinyl alcohol electrostatic spinning membrane as well as preparation method and application thereof

Abstract

The invention provides an antibacterial polyvinyl alcohol electrostatic spinning membrane as well as a preparation method and application thereof, the antibacterial polyvinyl alcohol electrostatic spinning membrane is prepared by taking polyvinyl alcohol as a spinning base material, ethylene oxide as a spinning aid and polyhexamethylene biguanide hydrochloride and benzalkonium chloride as double antibacterial agents through an electrostatic spinning method. According to the PVA nanofiber membrane, the double antibacterial agents PHMB and BAC are added into the PVA nanofiber membrane through the electrostatic spinning technology, the excellent biocompatibility characteristic of PVA is reserved, the PVA nanofiber membrane is of a three-dimensional porous structure, the antibacterial property is greatly improved, the PVA nanofiber membrane can be widely applied to the fields of tissue engineering, medical engineering and the like, and more technical supports are provided for biomedical application.

Description

Technical Field

[0001] The present application relates to the technical field of nanomaterials, and more specifically, to an antibacterial polyvinyl alcohol electrospinning membrane and a preparation method and application thereof. Background Art

[0002] Polyvinyl alcohol (PVA) is a water-soluble polymer. The film made of it is flexible, smooth, oil-resistant, solvent-resistant, abrasion-resistant, and has good gas barrier properties. It can be used as wound dressings, cell scaffolds, in vitro tissue reconstruction, etc. PVA electrospun membranes have a huge surface area to volume ratio, high porosity, sufficient permeability, excellent wound exudate absorption capacity, structural similarity with the extracellular matrix of the skin, excellent antibacterial properties, long-term sustained-release properties, and good biocompatibility. It has broad development prospects in the field of biomedicine.

[0003] Polyhexamethylene biguanide hydrochloride (PHMB) is a new, efficient, multi-purpose polymer, which belongs to cationic surfactant. Its polymer structure improves the effective activity of guanidine group, and its bactericidal effect is higher than other guanidines.

[0004] Benzalkonium chloride (BAC) is a cationic surfactant and a non-oxidizing bactericidal agent with a broad-spectrum and highly effective bactericidal ability. It can change the permeability of bacterial cytoplasmic membranes, causing the extravasation of bacterial cytoplasmic substances, hindering their metabolism and thus killing them. Summary of the invention

[0005] The present invention provides an antibacterial polyvinyl alcohol electrospun membrane and a preparation method and application thereof. By utilizing electrospinning technology, PHMB and BAC dual antibacterial agents are added to the PVA nanofiber membrane, thereby retaining the excellent biocompatibility characteristics of PVA and having a three-dimensional porous structure, greatly improving the antibacterial property. The membrane can be widely used in many fields such as tissue engineering and medical engineering, providing more technical support for biomedical applications.

[0006] In a first aspect, the present invention provides an antibacterial polyvinyl alcohol electrospun membrane, which is prepared by an electrospinning method using polyvinyl alcohol as a spinning substrate, ethylene oxide as a spinning aid, polyhexamethylene biguanide hydrochloride and benzalkonium chloride as dual antibacterial agents.

[0007] Furthermore, the antibacterial polyvinyl alcohol electrospinning membrane has a three-dimensional porous structure with uniform diameter and continuous fiber morphology.

[0008] Furthermore, the antibacterial polyvinyl alcohol electrospun membrane has an inhibitory effect on Escherichia coli and Staphylococcus aureus.

[0009] Furthermore, the diameter of the nanofibers of the antibacterial polyvinyl alcohol electrospun membrane is 190-210 nm. Preferably, the diameter of the nanofibers is 200 nm.

[0010] In a second aspect, the present invention provides a method for preparing an antibacterial polyvinyl alcohol electrospinning membrane, comprising the following steps: (1) mixing polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride and benzalkonium chloride, adding deionized water to obtain a mixed solution; (2) heating the mixed solution in a water bath and continuously stirring to obtain an electrospinning precursor solution; (3) loading the electrospinning precursor solution into a syringe, connecting the syringe to a spinning machine, setting spinning parameters, and performing electrospinning to obtain an electrospinning membrane; (4) vacuum drying the electrospinning membrane to obtain the antibacterial polyvinyl alcohol electrospinning membrane.

[0011] Furthermore, in step (1), the mass ratio of polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride and benzalkonium chloride is 100:20:1:5.

[0012] Further, in step (2), the water bath temperature is 60-80°C, and the stirring time is 12-24h. Preferably, in step (2), the water bath temperature is 80°C, and the stirring time is 24h.

[0013] Further, in step (3), the spinning voltage is set to 16-20 kV, the receiving distance is 18-22 cm, the injection speed is 0.6-1.0 mL / h, and the spinning time is 4-5 h. Preferably, in step (3), the spinning voltage is set to 16-18 kV, the receiving distance is 18-20 cm, the injection speed is 0.6-0.8 mL / h, and the spinning time is 4-5 h.

[0014] Furthermore, in step (4), the drying temperature is 50-80°C, and the drying time is 24-36 hours. Preferably, in step (4), the drying temperature is 60°C, and the drying time is 36 hours.

[0015] In a third aspect, the present invention provides the use of the antibacterial polyvinyl alcohol electrospinning membrane as described above in the fields of tissue engineering and medical engineering.

[0016] In summary, this application has the following beneficial effects:

[0017] 1. The preparation method of the present invention as a whole is closely related and inseparable from each other. In this system, the synergistic effect of the two antibacterial agents can significantly enhance the antibacterial performance and improve the nanofiber structure. Since the polyvinyl alcohol and polyethylene oxide used are non-toxic and have good biocompatibility, the prepared nanofiber membrane is environmentally friendly;

[0018] 2. The electrospinning equipment and preparation method of the present invention are simple and easy to operate, and the reaction conditions are easy to control at room temperature and pressure;

[0019] 3. The raw material polyvinyl alcohol used in the electrospinning membrane of the present invention is cheap, degradable and has good biocompatibility;

[0020] 4. In the present invention, polyhexamethylene biguanide hydrochloride and benzalkonium chloride are selected as antibacterial agents, and the synergistic effect of the two antibacterial agents is utilized to improve their antibacterial properties. The prepared electrospun membrane has good biocompatibility and obvious antibacterial effect.

[0021] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not intended to limit the scope of protection of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a scanning electron microscope image of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention.

[0023] Figure 2 This is a transmission electron micrograph of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention.

[0024] Figure 3 This is a graph showing the Escherichia coli inhibition rate of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention.

[0025] Figure 4 This is a graph showing the Staphylococcus aureus inhibition rate of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention.

[0026] Figure 5 This is a photograph of the Escherichia coli inhibition zone of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention.

[0027] Figure 6 This is a photograph of the Staphylococcus aureus inhibition zone of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention. DETAILED DESCRIPTION

[0028] The present application is further described in detail below in conjunction with the examples. It is particularly noted that if no specific conditions are specified in the following examples, the experiments are carried out according to conventional conditions or the conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following examples can be obtained from common commercial sources.

[0029] Example

[0030] Example 1: PVA / PEO / PHMB / BAC (double antimicrobial agent)

[0031] A method for preparing an antibacterial polyvinyl alcohol electrospinning membrane comprises the following steps:

[0032] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, 0.01 g of polyhexamethylene biguanide hydrochloride, and 0.05 g of benzalkonium chloride. Mix polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride, and benzalkonium chloride in a mass ratio of 100:20:1:5, and add 10 mL of deionized water to prepare an aqueous solution.

[0033] (2) The solution was heated in a water bath at 80°C and stirred continuously for 24 hours to obtain an electrospinning precursor solution with good uniformity.

[0034] (3) The precursor solution was loaded into a syringe and connected to a spinning machine. The spinning voltage was set to 18 kV, the receiving distance was set to 20 cm, and the injection speed was set to 0.8 mL / h. The electrospinning was performed for 5 h to obtain an electrospun membrane.

[0035] (4) The electrospun membrane was vacuum dried at 60°C for 36 h to obtain an antibacterial polyvinyl alcohol electrospun membrane.

[0036] Comparative Example 1: PVA / PEO (without antimicrobial agent)

[0037] (1) Weigh 1 g of polyvinyl alcohol and 0.2 g of polyethylene oxide. Mix the polyvinyl alcohol and polyethylene oxide in a mass ratio of 100:20, and add 10 mL of deionized water to prepare an aqueous solution.

[0038] Comparative Example 2: PVA / PEO / PHMB (single antimicrobial agent)

[0039] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, and 0.01 g of polyhexamethylene biguanide hydrochloride. Mix the polyvinyl alcohol, polyethylene oxide, and polyhexamethylene biguanide hydrochloride in a mass ratio of 100:20:1, and add 10 mL of deionized water to prepare an aqueous solution.

[0040] Comparative Example 3: PVA / PEO / BAC (single antimicrobial agent)

[0041] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, and 0.05 g of benzalkonium chloride. Mix the polyvinyl alcohol, polyethylene oxide, and benzalkonium chloride in a mass ratio of 100:20:5, and add 10 mL of deionized water to prepare an aqueous solution.

[0042] Comparative Example 1, Comparative Example 2 and Comparative Example 3 were used as spinning formulas, respectively, and the other spinning conditions were within the range. The subsequent experimental steps were the same, and electrospinning membranes were prepared respectively.

[0043] The performance effects of the antibacterial polyvinyl alcohol electrospinning membrane prepared by the present invention are as follows:

[0044] The antibacterial polyvinyl alcohol electrospun membrane obtained in this embodiment is a nanofiber (PVA / PEO / PHMB / BAC) with double antibacterial agents added, such as Figure 1 As shown, it can be seen that the diameter is uniform, about 200nm.

[0045] like Figure 2 As shown, it is shown that the antibacterial polyvinyl alcohol electrospun membrane obtained in this embodiment has uniform diameter, excellent fiber morphology, and no defects such as droplets and breaks, which is more conducive to the uniform release of the antibacterial agent and provides good and stable antibacterial performance.

[0046] Escherichia coli and Staphylococcus aureus were cultured respectively, and antibacterial tests were carried out using Example and Comparative Example 1, Comparative Example 2, and Comparative Example 3 respectively, and calculations were performed using absorbance values ​​to obtain Tables 1 and 2.

[0047] Table 1 Comparison of the antibacterial effect of Escherichia coli in Example 1 and Comparative Examples 1 to 3

[0048] Sample No. E. coli survival rate Example 1 (PVA / PEO / PHMB / BAC) 6.54％ Comparative Example 1 (PVA / PEO) 100％ Comparative Example 2 (PVA / PEO / PHMB) 11.21％ Comparative Example 3 (PVA / PEO / BAC) 9.21％

[0049] Table 1 is a comparison of the antibacterial rates of Escherichia coli in Example 1 and Comparative Examples 1 to 3. Figure 3 This is a graph showing the Escherichia coli inhibition rate of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention and the electrospun membranes in Comparative Examples 1, 2, and 3.

[0050] Table 2 Comparison of the antibacterial effect of Staphylococcus aureus in Example 1 and Comparative Examples 1 to 3

[0051]

[0052]

[0053] Table 2 is a comparison of the antibacterial rates of Staphylococcus aureus in Example 1 and Comparative Examples 1 to 3. Figure 4 The antibacterial rate of Staphylococcus aureus of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention and the electrospun membranes in Comparative Examples 1, 2, and 3 is shown in the figure. The vertical axis is the survival rate of bacteria. The lower the survival rate of bacteria, the better the antibacterial property of the formula. The corresponding formulas are marked below the horizontal axis for comparison of antibacterial properties.

[0054] From Table 1, Table 2, Figure 3 and Figure 4 It can be seen that the antibacterial polyvinyl alcohol electrospun membrane in this embodiment has a significantly higher antibacterial rate against Escherichia coli and Staphylococcus aureus than the electrospun membrane PVA / PEO without antibacterial agent and the electrospun membranes PVA / PEO / PHMB and PVA / PEO / BAC with added single antibacterial agent in Comparative Examples 1 to 3 due to the addition of dual antibacterial agents.

[0055] Figure 5 These are photos of the Escherichia coli inhibition zones of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention and the electrospun membranes in Comparative Examples 1, 2, and 3. Figure 6 The photos are of the Staphylococcus aureus inhibition zones of the antibacterial polyvinyl alcohol electrospun membrane in Example 1 of the present invention and the electrospun membranes in Comparative Examples 1, 2, and 3. Figure 5 and Figure 6 The formulas No. 1 to No. 4 are respectively: Comparative Example 1 (PVA / PEO), Comparative Example 2 (PVA / PEO / PHMB), Comparative Example 3 (PVA / PEO / BAC) and Example 1 (PVA / PEO / PH MB / BAC), and the inner circle is a nanofiber membrane. The nanofiber membranes of different formulas are placed on a culture dish full of bacteria. After constant temperature culture for the same time, the larger the outer annular area, the better the antibacterial performance.

[0056] like Figure 5 and Figure 6 As shown, it is shown that the inhibition zone of the antibacterial polyvinyl alcohol electrospun membrane in this embodiment for Escherichia coli and Staphylococcus aureus is significantly greater than that of the electrospun membrane PVA / PEO without antibacterial agent in Comparative Examples 1 to 3 and the electrospun membranes PVA / PEO / PHMB and PVA / PEO / BAC with added single antibacterial agent.

[0057] Example 2

[0058] A method for preparing an antibacterial polyvinyl alcohol electrospinning membrane comprises the following steps:

[0059] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, 0.01 g of polyhexamethylene biguanide hydrochloride, and 0.05 g of benzalkonium chloride. Mix polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride, and benzalkonium chloride in a mass ratio of 100:20:1:5, and add 10 mL of deionized water to prepare an aqueous solution.

[0060] (2) The solution was heated in a water bath at 80°C and stirred continuously for 24 hours to obtain an electrospinning precursor solution with good uniformity.

[0061] (3) The precursor solution was loaded into a syringe and connected to a spinning machine. The spinning voltage was set to 16 kV, the receiving distance was 18 cm, and the injection speed was 0.6 mL / h. The electrospinning was performed for 4 h to obtain an electrospun membrane.

[0062] (4) The electrospun membrane was vacuum dried at 80°C for 36 h to obtain an antibacterial polyvinyl alcohol electrospun membrane.

[0063] Example 3

[0064] A method for preparing an antibacterial polyvinyl alcohol electrospinning membrane comprises the following steps:

[0065] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, 0.01 g of polyhexamethylene biguanide hydrochloride, and 0.05 g of benzalkonium chloride. Mix polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride, and benzalkonium chloride in a mass ratio of 100:20:1:5, and add 10 mL of deionized water to prepare an aqueous solution.

[0066] (2) The solution was heated in a water bath at 80°C and stirred continuously for 24 hours to obtain an electrospinning precursor solution with good uniformity.

[0067] (3) The precursor solution was loaded into a syringe and connected to a spinning machine. The spinning voltage was set to 20 kV, the receiving distance was set to 22 cm, and the injection speed was set to 1.0 mL / h. The electrospinning was performed for 4 h to obtain an electrospun membrane.

[0068] (4) The electrospun membrane was vacuum dried at 50°C for 24 h to obtain an antibacterial polyvinyl alcohol electrospun membrane.

[0069] Example 4

[0070] A method for preparing an antibacterial polyvinyl alcohol electrospinning membrane comprises the following steps:

[0071] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, 0.01 g of polyhexamethylene biguanide hydrochloride, and 0.05 g of benzalkonium chloride. Mix polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride, and benzalkonium chloride in a mass ratio of 100:20:1:5, and add 10 mL of deionized water to prepare an aqueous solution.

[0072] (2) The solution was heated in a water bath at 80°C and stirred continuously for 12 hours to obtain an electrospinning precursor solution with good uniformity.

[0073] (3) The precursor solution was loaded into a syringe and connected to a spinning machine. The spinning voltage was set to 17 kV, the receiving distance was 18 cm, and the injection speed was 1.0 mL / h. The electrospinning was performed for 5 h to obtain an electrospun membrane.

[0074] (4) The electrospun membrane was vacuum dried at 60°C for 24 h to obtain an antibacterial polyvinyl alcohol electrospun membrane.

[0075] Example 5

[0076] A method for preparing an antibacterial polyvinyl alcohol electrospinning membrane comprises the following steps:

[0077] (1) Weigh 1 g of polyvinyl alcohol, 0.2 g of polyethylene oxide, 0.01 g of polyhexamethylene biguanide hydrochloride, and 0.05 g of benzalkonium chloride. Mix polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride, and benzalkonium chloride in a mass ratio of 100:20:1:5, and add 10 mL of deionized water to prepare an aqueous solution.

[0078] (2) The solution was heated in a water bath at 80°C and stirred continuously for 12 hours to obtain an electrospinning precursor solution with good uniformity.

[0079] (3) The precursor solution was loaded into a syringe and connected to a spinning machine. The spinning voltage was set to 19 kV, the receiving distance was set to 21 cm, and the injection speed was set to 0.9 mL / h. The electrospinning was performed for 4 h to obtain an electrospun membrane.

[0080] (4) The electrospun membrane was vacuum dried at 60°C for 36 h to obtain an antibacterial polyvinyl alcohol electrospun membrane.

[0081] The above is only an exemplary embodiment of the present disclosure, but the protection scope of the present disclosure is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present disclosure should be included in the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure should be based on the protection scope of the claims.

Claims

1. An antibacterial polyvinyl alcohol electrospinning membrane, characterized in that: The antibacterial polyvinyl alcohol electrostatic spinning membrane is prepared by an electrostatic spinning method using polyvinyl alcohol as a spinning substrate, ethylene oxide as a spinning aid, polyhexamethylene biguanide hydrochloride and benzalkonium chloride as dual antibacterial agents.

2. The antibacterial polyvinyl alcohol electrospinning membrane according to claim 1, characterized in that: The antibacterial polyvinyl alcohol electrospinning membrane has a three-dimensional porous structure with uniform diameter and continuous fiber morphology.

3. The antibacterial polyvinyl alcohol electrospinning membrane according to claim 1, characterized in that: The antibacterial polyvinyl alcohol electrospinning membrane has an inhibitory effect on Escherichia coli and Staphylococcus aureus.

4. The antibacterial polyvinyl alcohol electrospinning membrane according to claim 1, characterized in that: The diameter of the nanofibers of the antibacterial polyvinyl alcohol electrospinning membrane is 190-210 nm.

5. The method for preparing the antibacterial polyvinyl alcohol electrospinning membrane according to claim 1, characterized in that: The following steps are involved: (1) mixing polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride and benzalkonium chloride, and adding deionized water to obtain a mixed solution; (2) heating the mixed solution in a water bath and continuously stirring to obtain an electrospinning precursor solution; (3) loading the electrospinning precursor solution into a syringe, connecting the syringe to a spinning machine, setting spinning parameters, and performing electrospinning to obtain an electrospinning membrane; (4) vacuum drying the electrospun membrane to obtain the antibacterial polyvinyl alcohol electrospun membrane.

6. The method for preparing the antibacterial polyvinyl alcohol electrospinning membrane according to claim 5, characterized in that: In step (1), the mass ratio of the polyvinyl alcohol, polyethylene oxide, polyhexamethylene biguanide hydrochloride and benzalkonium chloride is 100:20:1:

5.

7. The method for preparing the antibacterial polyvinyl alcohol electrospinning membrane according to claim 5, characterized in that: In step (2), the water bath temperature is 60-80° C. and the stirring time is 12-24 h.

8. The method for preparing the antibacterial polyvinyl alcohol electrospinning membrane according to claim 5, characterized in that: In step (3), the spinning voltage is set to 16-20 kV, the receiving distance is set to 18-22 cm, the injection speed is set to 0.6-1.0 mL / h, and the spinning time is set to 4-5 h.

9. The method for preparing the antibacterial polyvinyl alcohol electrospinning membrane according to claim 5, characterized in that: In step (4), the drying temperature is 50-80°C and the drying time is 24-36h.

10. Use of the antibacterial polyvinyl alcohol electrospun membrane according to any one of claims 1 to 4 in the fields of tissue engineering and medical engineering.

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