CN110787316A - A kind of AIE composite electrospinning fiber membrane and its preparation method and application - Google Patents

Abstract

The present invention provides an AIE composite electrospinning fiber membrane and a preparation method and application thereof. The AIE composite electrospinning fiber membrane is a nanofiber membrane formed by a biocompatible polymer and AIE molecules, which can be slowly Releases antibacterial active ingredient AIE molecules, effectively inhibits and kills bacteria, does not produce any side effects on normal cells, and can promote cell adhesion, growth and proliferation, has good antibacterial and breathable properties, and can completely fit in wounds The surface maintains a stable physiological environment and accelerates wound healing. The AIE composite electrospinning fiber membrane is prepared by electrospinning technology, the preparation method is simple, the cost is low, it is suitable for large-scale industrial production, and has important application value in the treatment of chronic wounds such as wounds or burns.

Description

A kind of AIE composite electrospinning fiber membrane and its preparation method and application

technical field

The invention belongs to the technical field of biomedical materials, and in particular relates to an AIE composite electrospinning fiber membrane and a preparation method and application thereof.

Background technique

Chronic wounds such as burns and wounds are highly susceptible to bacterial infection, which can lead to infection-related complications such as sepsis and acute renal failure, especially after the emergence of super-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA). become an almost unavoidable clinical problem. According to the statistics of the World Health Organization, more than 25 million people in the world suffer from traumatic infection every year, and the cost of treatment for wound infection is more than tens of billions of dollars. Wound infection has become a major problem that threatens human health in the future. question. Not only that, with the abuse of antibiotics, the mutation frequency of bacteria has accelerated significantly, and there are more and more resistant mutant strains. The continuous evolution of super-resistant bacteria has made human beings particularly urgent for the research and development of new antibacterial drugs and antibacterial materials. .

In the treatment of chronic wounds such as burns and wounds, the systemic use of antibiotics often results in that the local drug concentration in the wound cannot reach the effective range for preventing and treating infection, and increasing the drug dose will bring certain side effects to patients. Therefore, using Appropriate topical antibacterial dressing to achieve the purpose of reducing wound infection and promoting wound healing is the consensus of the current clinical community.

In recent years, a variety of dressings with antibacterial properties have been developed, which are mainly divided into fibers, films, hydrogels and sponges according to their carrier forms. The antibacterial actives loaded on the dressings include natural antibacterial molecules, traditional antibiotics and their derivatives. substances, antibacterial polymers, and antibacterial nanoparticles. For example, CN104740676A discloses a procyanidin cross-linked gelatin antibacterial dressing and a preparation method thereof. The preparation method is as follows: casting a certain concentration of gelatin solution into a mold to freeze-dry to obtain a gelatin sponge stent, and then soaking the gelatin sponge stent in the procyanidin solution Cross-linking is carried out, and an antibiotic solution is added at the same time, so that small antibiotic molecules enter the gelatin, and freeze-drying is performed again to obtain the antibacterial dressing. The material has good physical and chemical properties and biocompatibility, and can be used for the repair and replacement of wounds in the medical field. CN105709262A discloses a silver-loaded antibacterial dressing and a preparation method thereof. The preparation method is as follows: respectively dissolving a silver salt component and a stabilizer component in water to obtain a silver salt antibacterial solution, and then immersing a fiber and other antibacterial dressing substrates or Padding in a silver salt antibacterial solution and drying to obtain a silver-loaded antibacterial dressing. The silver-loaded antibacterial dressing has excellent antibacterial properties, and the release of silver ions is stable, and it is not easy to cause silver shedding and pigment deposition in wounds. CN103920180A discloses a chitosan hydrogel for antibacterial dressings and a preparation method thereof. The preparation method comprises the following steps: introducing carboxyl groups on chitosan by means of free radical polymerization, and the carboxyl groups on the chitosan and the antibacterial agent polyhexamethylene The amidation reaction of methyl guanidine hydrochloride is carried out, and polyhexamethylene guanidine hydrochloride is grafted on chitosan to obtain chitosan for antibacterial dressing, and then cross-linked to obtain the chitosan hydrogel for antibacterial dressing. The chitosan hydrogel for antibacterial dressings has the characteristics of good drug resistance, good antibacterial properties, durable antibacterial properties, high water absorption rate and good mechanical properties.

However, in the existing antibacterial dressings, antibiotics and their derivatives take effect quickly, but cross-resistance is prone to occur; antibacterial polymers have strong side effects and poor solubility, so they cannot be directly used for the treatment of infectious diseases; nano silver, etc. The stability, toxicity and in vivo drug metabolism mechanism of antibacterial particles have not been fully clarified, and further research is needed.

Therefore, developing a novel high-efficiency antibacterial material to inhibit wound bacteria and promote wound healing is the focus of research in this field.

SUMMARY OF THE INVENTION

In view of the deficiencies of the prior art, the purpose of the present invention is to provide an AIE composite electrospinning fiber membrane and a preparation method and application thereof. The AIE composite electrospinning fiber membrane is loaded with AIE molecules with antibacterial function, which can inhibit bacteria It can grow and accelerate wound healing, and has important application value in the treatment of chronic wounds such as wounds or burns.

In order to achieve this object of the invention, the present invention adopts the following technical solutions:

In a first aspect, the present invention provides an AIE composite electrospinning fiber membrane, which is an electrospinning fiber membrane formed by a biocompatible polymer and AIE molecules.

The AIE composite electrospinning fiber membrane provided by the present invention is loaded with AIE molecules, and the AIE is an aggregation-induced emission material (Aggregation-induced emission), which can enhance fluorescence in the aggregation state, has good biocompatibility and low background fluorescence , good photostability and other characteristics, it has shown great application prospects in many fields such as biological detection, biological imaging and integration of diagnosis and treatment. Through research, the present invention finds that AIE molecules with multiple conjugated rigid arm structures can insert their conjugated rigid arm structures into the cell wall of bacteria, and achieve excellent antibacterial performance by inhibiting the synthesis of the bacterial cell wall.

On the one hand, the AIE composite electrospinning fiber membrane of the present invention has excellent antibacterial function based on the introduction of AIE molecules; on the other hand, the electrospinning nanofiber membrane has large specific surface area and high porosity, and is a The three-dimensional network structure similar to the human extracellular matrix structure is beneficial to promote cell adhesion, growth and proliferation, and accelerate wound healing; at the same time, the biocompatible polymer in the AIE composite electrospinning fiber membrane has good properties. Biocompatibility and blood compatibility, it is an ideal biomedical raw material. Therefore, the AIE composite electrospinning fiber membrane provided by the present invention cooperates with each other through the biocompatible polymer, the antibacterial AIE molecule and the spatial three-dimensional structure of the electrospinning fiber membrane, which together endow the AIE composite electrospinning fiber with each other. The film's good antibacterial properties and wound-healing-promoting properties make it of great clinical significance and application value in the treatment of chronic wounds.

Preferably, the AIE molecule is selected from tetrastyrene, 1,1,2,2-tetra-[4-carboxy-(1,1-biphenyl)]ethylene or tetra-(4-hydroxyphenyl)ethylene any one or a combination of at least two.

Preferably, the biocompatible polymer is selected from polylactic acid, polylactic acid-glycolic acid copolymer, polycaprolactone, lactide-caprolactone copolymer, sodium alginate, chitosan or hyaluronic acid any one or a combination of at least two.

Preferably, the mass ratio of the biocompatible polymer and the AIE molecule is 1:(0.001-0.1), for example, 1:0.002, 1:0.004, 1:0.005, 1:0.007, 1:0.009, 1:0.01 , 1:0.02, 1:0.03, 1:0.04, 1:0.05, 1:0.06, 1:0.07, 1:0.08, 1:0.09, or 1:0.095, etc., more preferably 1:(0.01-0.08).

In the present invention, when the mass ratio of the biocompatible polymer and AIE molecules is 1:(0.001-0.1), the obtained AIE composite electrospinning fiber membrane has excellent antibacterial properties. If the content of AIE molecules is too low , the bacteriostatic effect of the AIE composite electrospinning fiber membrane is weakened, and the effect of bacteriostasis and wound healing cannot be achieved; if the content of AIE molecules is too high, it will cause waste of antibacterial active ingredients.

Preferably, the fiber diameter of the AIE composite electrospinning fiber membrane is 100-800 nm, such as 120 nm, 150 nm, 170 nm, 200 nm, 230 nm, 250 nm, 280 nm, 300 nm, 330 nm, 350 nm, 380 nm, 400 nm, 420 nm, 450 nm, 480 nm , 500nm, 520nm, 550nm, 570nm, 600nm, 630nm, 650nm, 680nm, 700nm, 720nm, 750nm, 770nm or 790nm, etc.

In the present invention, when the fiber diameter of the AIE composite electrospinning fiber membrane is 100-800 nm, an electrospinning fiber membrane with high porosity and large specific surface area can be obtained, so that the AIE composite electrospinning fiber membrane can be The three-dimensional structure of space is similar to the structure of human extracellular matrix, which is beneficial to promote cell adhesion and growth and accelerate wound healing. If the diameter of the fiber exceeds the above range, the obtained electrospinning fiber membrane cannot mimic the human extracellular matrix and cannot achieve the purpose of promoting cell growth.

On the other hand, the present invention provides a preparation method of the above-mentioned AIE composite electrospinning fiber membrane, and the preparation method comprises the following steps:

(1) AIE molecule is mixed with organic solvent to obtain AIE solution;

(2) mixing the biocompatible polymer with an organic solvent to obtain an electrospinning precursor;

(3) mixing and dispersing the AIE solution obtained in step (1) and the electrospinning precursor solution obtained in step (2) to obtain an electrospinning solution;

(4) Electrospinning the electrospinning solution obtained in step (3) to obtain the AIE composite electrospinning fiber membrane.

Preferably, the organic solvent in step (1) is selected from any one or a combination of at least two of dimethyl sulfoxide, N,N-dimethylformamide or acetone.

Preferably, the concentration of AIE molecules in the AIE solution of step (1) is 0.5-100 mg/mL, such as 0.6 mg/mL, 0.8 mg/mL, 1 mg/mL, 3 mg/mL, 5 mg/mL, 8 mg/mL, 10mg/mL, 15mg/mL, 20mg/mL, 25mg/mL, 30mg/mL, 35mg/mL, 40mg/mL, 45mg/mL, 50mg/mL, 55mg/mL, 60mg/mL, 65mg/mL, 70mg/mL mL, 75mg/mL, 80mg/mL, 85mg/mL, 90mg/mL, 95mg/mL or 99mg/mL etc.

Preferably, the mixing in step (1) is carried out under ultrasonic conditions.

Preferably, the frequency of the ultrasound is 20-50 kHz, such as 22 kHz, 25 kHz, 27 kHz, 30 kHz, 33 kHz, 35 kHz, 38 kHz, 40 kHz, 42 kHz, 45 kHz, 47 kHz or 49 kHz.

Preferably, the power of the ultrasound is 100-200W, such as 105W, 110W, 115W, 120W, 125W, 130W, 135W, 140W, 145W, 150W, 155W, 160W, 165W, 170W, 175W, 180W, 185W, 190W or 195W, etc.

Preferably, the ultrasonic time is 5 to 60 minutes, such as 8 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, or 58 minutes.

Preferably, the AIE solution in step (1) is stored in the dark.

Preferably, the organic solvent in step (2) is selected from acetone, N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, chloroform, anhydrous ethanol, tetrahydrofuran or hexafluoroisopropanol any one or a combination of at least two.

Preferably, the dispersion in step (3) is carried out under a dark condition.

Preferably, the dispersion time in step (3) is 2-12h, for example, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, 5.5h, 6h, 6.5h, 7h, 7.5h, 8h, 8.5h , 9h, 9.5h, 10h, 10.5h, 11h or 11.5h, etc.

Preferably, the mass percentage content of the biocompatible polymer in the electrospinning solution in step (3) is 10-30%, such as 12%, 14%, 15%, 17%, 19%, 20%, 22%, 24%, 25%, 27% or 29% etc.

Preferably, the mass ratio of AIE molecules to biocompatible polymers in the electrospinning solution in step (3) is (0.001-0.1):1, for example, 0.002:1, 0.004:1, 0.005:1, 0.007: 1, 0.009:1, 0.01:1, 0.015:1, 0.02:1, 0.03:1, 0.04:1, 0.05:1, 0.06:1, 0.07:1, 0.08:1, 0.09:1 or 0.095:1, etc. .

Preferably, the parameters of the electrospinning in step (4) are set as follows: the bolus injection rate of the electrospinning solution is 0.2-2mL/h (for example, 0.3mL/h, 0.5mL/h, 0.7mL/h, 0.9mL /h, 1mL/h, 1.2mL/h, 1.4mL/h, 1.6mL/h, 1.8mL/h or 1.9mL/h, etc.), the loading voltage is 10~20kV (such as 11kV, 12kV, 13kV, 14kV, 15kV, 16kV, 17kV, 18kV or 19kV, etc.), the receiving distance is 10-25cm (such as 11cm, 13cm, 15cm, 17cm, 19cm, 20cm, 22cm or 24cm, etc.), the diameter of the spinning nozzle is 0.3-1mm (such as 0.4 mm, 0.5mm, 0.6mm, 0.7mm, 0.8mm or 0.9mm, etc.).

Preferably, the preparation method specifically comprises the following steps:

(1) Dissolving AIE molecules in an organic solvent under the assistance of ultrasound to obtain an AIE solution with an AIE molecular concentration of 0.5-100 mg/mL, and the AIE solution is protected from light;

(2) dissolving the biocompatible polymer in an organic solvent to obtain an electrospinning precursor solution with a static mass percentage of the biocompatible polymer of 10-30%;

(3) Mixing the AIE solution obtained in step (1) with the electrospinning precursor solution obtained in step (2), avoiding light and dispersing evenly, to obtain a mass ratio of AIE molecule to biocompatible polymer (0.001～0.1): 1 of the electrospinning solution;

(4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as: the injection speed of the electrospinning solution is 0.2-2 mL/h, and the loading voltage is 10-20 kV , the receiving distance is 10-25 cm, the diameter of the spinning nozzle is 0.3-1 mm, and the AIE composite electrospinning fiber membrane is obtained.

In another aspect, the present invention provides an application of the above-mentioned AIE composite electrospinning fiber membrane in the preparation of biomedical materials.

In another aspect, the present invention provides an antibacterial dressing comprising the above-mentioned AIE composite electrospinning fiber film.

Compared with the prior art, the present invention has the following beneficial effects:

The AIE composite electrospinning fiber film provided by the present invention is an electrospinning fiber film with a three-dimensional network structure that uses a biocompatible polymer as a base material and is loaded with antibacterial AIE molecules. The fiber membrane can slowly release the antibacterial active ingredient AIE molecule during use, which can effectively inhibit and kill pathogenic bacteria including Staphylococcus aureus and methicillin-resistant Staphylococcus aureus without any side effects on normal cells. , and can promote the adhesion, growth and proliferation of cells, has good antibacterial and breathable properties, can completely fit on the surface of the wound to maintain a stable physiological environment, and accelerate the healing speed of the wound. The AIE composite electrospinning fiber membrane is prepared by electrospinning technology, the preparation method is simple, the cost is low, it is suitable for large-scale industrial production, and has important application value in the treatment of chronic wounds such as wounds or burns.

Description of drawings

Fig. 1 is the scanning electron microscope image of the AIE composite electrospinning fiber membrane provided in Example 1;

Fig. 2 is the scanning electron microscope image of the AIE composite electrospinning fiber membrane provided in Example 2;

3 is a cell survival test chart of the AIE composite electrospinning fiber membrane provided in Example 1;

4 is a cell survival test chart of the AIE composite electrospinning fiber membrane provided in Example 2;

Fig. 5 is the cell morphology test chart on the AIE composite electrospinning fiber membrane provided in Example 1;

Fig. 6 is the cell morphology test chart on the AIE composite electrospinning fiber membrane provided in Example 2;

7 is a comparison diagram of the antibacterial properties of the AIE composite electrospinning fiber membranes provided by Example 1, Example 5, and Comparative Example 1;

FIG. 8 is a comparison diagram of the antibacterial properties of the AIE composite electrospinning fiber membranes provided by Example 1, Example 5, and Comparative Example 1;

FIG. 9 is a graph of the morphology test of Staphylococcus aureus on the AIE composite electrospinning fiber membrane provided in Example 1. FIG.

Detailed ways

The technical solutions of the present invention are further described below through specific embodiments. It should be understood by those skilled in the art that the embodiments are only for helping the understanding of the present invention, and should not be regarded as a specific limitation of the present invention.

The experimental materials used in the following examples of the present invention include:

(1) Biocompatible polymers: polycaprolactone (PCL) with a molecular weight of 80,000 g/mol; polylactic acid-glycolic acid copolymer (PLGA) with a molecular weight of 120,000 g/mol; polylactic acid (PLA) with a molecular weight of 100,000 g /mol, lactide-caprolactone copolymer (PLCL), molecular weight is 110000 g/mol.

(2) AIE molecule: tetraphenylethylene, 1,1,2,2-tetra-[4-carboxy-(1,1-biphenyl)]ethylene and tetra-(4-hydroxyphenyl)ethylene were purchased from Xuzhou Dayang Biochemical Technology Co., Ltd.

Example 1

The present embodiment provides an AIE composite electrospinning fiber membrane, and the specific preparation method is as follows:

(1) Add 20 mg of 1,1,2,2-tetra-[4-carboxy-(1,1-biphenyl)]ethylene particles to 7 mL of N,N-dimethylformamide, and sonicate for 10 min to make AIE particles Completely dissolving to obtain an AIE solution with an AIE molecular concentration of 2.86 mg/mL, and the AIE solution is protected from light;

(2) Dissolve 2g PCL in 7mL of tetrahydrofuran, stir magnetically for 3h at room temperature until completely dissolved, to obtain an electrospinning precursor solution;

(3) adding the AIE solution obtained in step (1) to the electrospinning precursor solution obtained in step (2), stirring in the dark for 2 hours to completely dissolve it, and then standing for 0.5 hours to obtain a uniform and stable electrospinning solution Silk solution, the mass percentage of PCL in the electrospinning solution is 13%;

(4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as: the injection speed of the electrospinning solution is 1 mL/h, the loading voltage is 15 kV, and the receiving distance is 10 cm, the diameter of the spinning nozzle is 0.7 mm, to obtain the AIE composite electrospinning fiber membrane, and the average fiber diameter of the AIE composite electrospinning fiber membrane is 300 nm.

Example 2

The present embodiment provides an AIE composite electrospinning fiber membrane, and the specific preparation method is as follows:

(1) Add 50 mg of 1,1,2,2-tetra-[4-carboxy-(1,1-biphenyl)]ethylene particles to 2 g of N,N-dimethylformamide, and sonicate for 10 min to make AIE particles Completely dissolved to obtain an AIE solution with an AIE molecular concentration of 23.8 mg/mL, and the AIE solution is protected from light;

(2) Mix 1g of PLGA with 1g of N,N-dimethylformamide and 1g of acetone, and stir magnetically for 3h at room temperature to completely dissolve to obtain an electrospinning precursor solution;

(3) adding the AIE solution obtained in step (1) to the electrospinning precursor solution obtained in step (2), stirring in the dark for 2 hours to completely dissolve it, and then standing for 0.5 hours to obtain a uniform and stable electrospinning solution Silk solution, the mass percentage of PLGA in the electrospinning solution is 20%;

(4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as follows: the bolus injection speed of the electrospinning solution is 0.8 mL/h, the loading voltage is 20 kV, and the receiving distance is is 15 cm, the diameter of the spinning nozzle is 0.7 mm, and the AIE composite electrospinning fiber membrane is obtained, and the average fiber diameter of the AIE composite electrospinning fiber membrane is 500 nm.

Example 3

The present embodiment provides an AIE composite electrospinning fiber membrane, and the specific preparation method is as follows:

(1) 5 mg of tetraphenylethylene particles were added to 10 mL of dimethyl sulfoxide solvent, and the tetraphenylethylene was completely dissolved by ultrasonics for 10 min to obtain an AIE solution with an AIE molecular concentration of 0.5 mg/mL, and the AIE solution was protected from light;

(2) Dissolve 2 g of PLA in 5 mL of dimethyl sulfoxide, and stir magnetically for 3 h at room temperature to completely dissolve to obtain an electrospinning precursor solution;

(3) adding the AIE solution obtained in step (1) to the electrospinning precursor solution obtained in step (2), stirring overnight in the dark to completely dissolve it, and then standing for 0.5 h to obtain a uniform and stable electrospinning solution Silk solution, the mass percentage of PLA in the electrospinning solution is 11%;

(4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as follows: the injection speed of the electrospinning solution is 0.2 mL/h, the loading voltage is 10 kV, and the receiving distance is is 10 cm, the diameter of the spinning nozzle is 0.3 mm, and the AIE composite electrospinning fiber film is obtained, and the average fiber diameter of the AIE composite electrospinning fiber film is 100 nm.

Example 4

The present embodiment provides an AIE composite electrospinning fiber membrane, and the specific preparation method is as follows:

(1) Mix 360 mg of tetra-(4-hydroxyphenyl) ethylene particles with 2 mL of N,N-dimethylformamide and 1.5 mL of acetone, and ultrasonicate for 10 min to completely dissolve tetraphenylethylene to obtain an AIE molecular concentration of 0.5 mg/mL mL of AIE solution, the AIE solution is protected from light;

(2) Dissolve 4.5g PLCL in 7mL N,N-dimethylformamide, stir magnetically for 3h at room temperature until completely dissolved, to obtain an electrospinning precursor solution;

(3) adding the AIE solution obtained in step (1) to the electrospinning precursor solution obtained in step (2), stirring overnight in the dark to completely dissolve it, and then standing for 1 hour to obtain uniform and stable electrospinning liquid, the mass percentage of chitosan in the electrospinning liquid is 30%;

(4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as: the bolus injection speed of the electrospinning solution is 2mL/h, the loading voltage is 15kV, and the receiving distance is 25 cm, the diameter of the spinning nozzle is 1 mm, to obtain the AIE composite electrospinning fiber membrane, and the average fiber diameter of the AIE composite electrospinning fiber membrane is 800 nm.

Example 5

The difference between this example and Example 1 is that the amount of AIE particles added in step (1) is 100 mg.

Comparative Example 1

This comparative example provides an electrospinning fiber membrane, and the specific preparation method is as follows:

(1) Dissolve 2 g of PCL in 7 mL of tetrahydrofuran, stir magnetically for 3 h at room temperature until completely dissolved, to obtain an electrospinning precursor solution;

(3) adding 7 mL of N,N-dimethylformamide to the electrospinning precursor solution obtained in step (1), stirring and mixing to obtain a uniform and stable electrospinning solution, in which PCL The mass percentage of 13%;

(4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as follows: the bolus injection speed of the electrospinning solution is 0.2-2 mL/h, the loading voltage is 15 kV, and the receiving voltage is 15 kV. The distance is 10 cm, the diameter of the spinning nozzle is 0.7 mm, and the electrospinning fiber film is obtained, and the average fiber diameter of the electrospinning fiber film is 300 nm.

Comparative Example 2

The difference between this comparative example and Example 1 is only that the 1,1,2,2-tetra-[4-carboxy-(1,1-biphenyl)]ethylene particles in step (1) are used -(4-Bromophenyl)ethylene particle replacement.

Comparative Example 3

The only difference between this comparative example and Example 1 is that the amount of 1,1,2,2-tetra-[4-carboxy-(1,1-biphenyl)]ethylene particles added in step (1) is 300 mg.

Test Example 1

This test example is a morphology test experiment of AIE composite electrospinning fiber membrane, and the specific method is as follows:

Scanning electron microscope (SEM, SU8200 type) was used to test the morphology of the AIE composite electrospinning fiber membranes provided in Examples 1-5 and Comparative Examples 1-3 of the present invention. Exemplarily, the scanning electron microscope image of the AIE composite electrospinning fiber membrane in Example 1 is shown in FIG. 1 , the AIE composite electrospinning fiber membrane has a uniform three-dimensional network structure, and its average fiber diameter is 300 nm; Example The scanning electron microscope image of the AIE composite electrospinning fiber membrane in 2 is shown in FIG. 2 , the AIE composite electrospinning fiber membrane has a uniform three-dimensional network structure, and its average fiber diameter is 500 nm.

Test case 2

This test example is a cell survival test experiment on the AIE composite electrospinning fiber membrane. The specific methods are as follows:

(1) The AIE composite electrospinning fiber membrane provided in Example 1 was cut into a rectangle of ⁴ cm×5 cm, soaked in 75% ethanol for 30 min to sterilize, and washed twice with PBS to remove residues after drying; The 3T3 cells were seeded on the AIE composite electrospinning fiber membrane at a density of /cm ² , and after culturing for 3 days at 37 °C in a 5% _CO environment, the 3T3 cells on the AIE composite electrospinning fiber membrane were slowly washed with PBS, and then washed with PBS. The LIVE/DEAD live-dead staining kit was used for staining, and then the cells were observed and photographed under a single-photon laser confocal microscope. The surface of the AIE composite electrospinning fiber membrane has a good growth shape, and the AIE composite electrospinning fiber membrane is beneficial to the proliferation and spreading of cells.

(2) The AIE composite electrospinning fiber membrane provided in Example 2 was cut into a rectangle of ⁴ cm×5 cm, soaked in 75% ethanol for 30 min to sterilize, and washed twice with PBS to remove residues after drying; The HUVEC cells were seeded on the AIE composite electrospinning fiber membrane at a density of /cm ² , and after culturing for 3 days at 37 °C in a 5% _CO environment, the HUVEC cells on the AIE composite electrospinning fiber membrane were slowly washed with PBS, and the The LIVE/DEAD live and dead staining kit was stained, and then observed and photographed under a single-photon laser confocal microscope. The obtained cell survival test chart is shown in Figure 4. It can be seen from Figure 4 that HUVEC cells were electrospun in AIE composite electrospinning. The growth morphology on the surface of the fiber membrane is good, and the AIE composite electrospinning fiber membrane in Example 2 is beneficial to the proliferation and spreading of HUVEC cells.

Test case 3

This test example is a cell morphology test experiment on the AIE composite electrospinning fiber membrane. The specific methods are as follows:

(1) According to the method of step (1) in Test Example 2, 3T3 cells were seeded on the AIE composite electrospinning fiber membrane provided in Example 1, and after culturing for 3 days at 37° C. in a 5% CO ₂ environment, gently use PBS Washed 3 times, fixed with prepared 4% paraformaldehyde, and placed in a refrigerator at 4°C overnight; then washed 3 times with distilled water to remove paraformaldehyde residue; used alcohol gradient dehydration replacement, the alcohol concentration was 30%, 50%, 70%, 80%, and 90% were dehydrated successively, and 100% was dehydrated three times for 10 min each time, to fully replace the water in the cells. The samples were dried in a vacuum freeze dryer for 1 h, then fixed on the sample stage, and used for secondary electron imaging with gold spray for 60 s before SEM observation to obtain the cell shape on the AIE composite electrospinning fiber membrane in Example 1. The appearance test diagram is shown in Fig. 5. It can be seen from Fig. 5 that the 3T3 cells have good cell morphology and growth state on the AIE composite electrospinning fiber membrane provided by the present invention.

(2) According to the method of step (2) in Test Example 2, HUVEC cells were seeded on the AIE composite electrospinning fiber membrane provided in Example 2, and after culturing for 3 days at 37° C., 5% CO ₂ environment, gently use PBS Washed 3 times, fixed with prepared 4% paraformaldehyde, and placed in a refrigerator at 4°C overnight; then washed 3 times with distilled water to remove paraformaldehyde residue; used alcohol gradient dehydration replacement, the alcohol concentration was 30%, 50%, 70%, 80%, and 90% were dehydrated successively, and 100% was dehydrated three times for 10 min each time, to fully replace the water in the cells. The samples were dried in a vacuum freeze dryer for 1 h, then fixed on the sample stage, and used for secondary electron imaging with gold spray for 60 s before SEM observation to obtain the cell shape on the AIE composite electrospinning fiber membrane in Example 2. The appearance test chart is shown in Figure 6. It can be seen from Figure 6 that the HUVEC cells have good cell morphology and growth state on the AIE composite electrospinning fiber membrane provided by the present invention.

Test Example 4

This test example is a cell proliferation activity test experiment on the AIE composite electrospinning fiber membrane. The specific methods are as follows:

Cell counting Kit-8 (CCK-8) was used to evaluate the cytotoxicity of the AIE composite electrospinning fiber membranes provided in Examples 1 to 5 and Comparative Examples 1 to 3 of the present invention. First, the AIE composite electrospinning fiber membranes were sterilized, After washing treatment, human umbilical vein endothelial cells (HUVEC) were seeded on AIE composite electrospinning fiber membranes at a density of 1 × 10 ⁴ /cm ² and cultured in a 37°C, 5% CO ₂ incubator for 24 hours. The cells were stained with CCK-8 and incubated for 2 hours, and the optical density of cells at 450 nm was measured with a multi-plate reader to obtain the cell viability (%) of HUVEC cells on the AIE composite electrospinning fiber membrane.

The cell viability on the AIE composite electrospinning fiber membranes provided in Examples 1-5 and Comparative Examples 1-3 was respectively tested according to the above-mentioned experimental methods, and the obtained test data are shown in Table 1.

Table 1

As can be seen from the data in Table 1, the AIE composite electrospinning fiber membranes provided in Examples 1 to 5 of the present invention have good biosafety and have no effect on cell growth and proliferation; the AIE composite electrospinning fiber membranes provided in Comparative Example 3 In the fibrous membrane, the mass ratio of AIE molecules to biocompatible polymers is 0.15:1, which is beyond the preferred mass range of the present invention, resulting in excessive AIE molecules and affecting the cell viability of normal fibroblasts.

Test Example 5

This test example is the antibacterial performance test experiment of AIE composite electrospinning fiber membrane, and the specific method is as follows:

(1) Add 100 μL of Staphylococcus aureus (S. aureus) bacterial solution to a 96-well plate, and test the OD value with a microplate reader; dilute the concentration of the bacterial solution to 10 ⁵ /mL with LB medium, and draw 10 μL of the bacterial solution Inoculated on the AIE composite electrospinning fiber membranes provided in Example 1, Example 5, and Comparative Example 1, respectively, cultivated in an incubator at 37°C for 24 hours, and then put the mixed culture medium into a 24-well plate, using LB The culture medium was used to prepare bacterial liquid with a concentration of 10 ⁴ /mL, and 100 μL of bacterial liquid was taken into a solid culture plate and spread evenly with a coating rod; it was placed upside down in a 37°C incubator for 24 hours, and the plate was taken out to count the colonies and take pictures. The comparison chart of the antibacterial properties of the AIE composite electrospinning fiber membranes in Example 1, Example 5, and Comparative Example 1 is shown in Figure 7. It can be seen from Figure 7 that the PCL electrospinning without the antibacterial active ingredient AIE molecule The silk fiber membrane (Comparative Example 1) has almost no antibacterial properties. The AIE composite electrospinning fiber membranes containing AIE molecules in Examples 1 and 5 of the present invention have obvious antibacterial properties, and the antibacterial properties in Example 5 are excellent. In Example 1, it was proved that with the increase of the AIE concentration in the electrospun fiber membrane, its antibacterial performance was further significantly improved.

(2) Add 100 μL of methicillin-resistant Staphylococcus aureus (MRSA) bacterial solution to a 96-well plate, and test the OD value with a microplate reader; dilute the concentration of the bacterial solution to 10 ⁵ /mL with LB medium, and draw 10 μL The bacterial liquid was inoculated on the AIE composite electrospinning fiber membranes provided in Example 1, Example 5 and Comparative Example 1, respectively, and cultivated in an incubator at 37°C for 24 hours, and then the mixed medium was placed in a 24-well plate. Use LB medium to prepare bacterial liquid with a concentration of 10 ⁴ /mL, take 100 μL of bacterial liquid and transfer it to a solid culture plate and spread it evenly with a coating rod; put it upside down in an incubator at 37 ° C for 24 hours, take out the plate to count the colonies and Taking pictures, the comparison chart of the antibacterial properties of the AIE composite electrospinning fiber membranes in Example 1, Example 5, and Comparative Example 1 is shown in Figure 8. It can be seen from Figure 8 that the PCL does not contain the antibacterial active ingredient AIE molecule. The electrospinning fiber membrane (Comparative Example 1) has almost no antibacterial properties, and the AIE composite electrospinning fiber membranes containing AIE molecules in Examples 1 and 5 of the present invention have obvious antibacterial properties against the super drug-resistant bacteria MRSA, and The antibacterial performance in Example 5 is better than that in Example 1. With the increase of AIE concentration in the electrospinning fiber film, the antibacterial performance of the prepared AIE composite electrospinning fiber film against super-resistant bacteria is further significantly improved.

According to the above experimental steps, the AIE composite electrospinning fiber membranes provided in Examples 1-5 and Comparative Examples 1-3 were tested against Staphylococcus aureus (S. aureus) and methicillin-resistant Staphylococcus aureus by plate colony counting method. (MRSA) antibacterial rate, the test data obtained are shown in Table 2.

Table 2

As can be seen from the data in Table 2, compared with the ordinary PCL electrospinning fiber membrane (Comparative Example 1) that does not contain the antibacterial active ingredient AIE molecule, the AIE composite electrospinning fiber membrane provided in Examples 1 to 5 of the present invention has a golden yellow color. Staphylococcus and super-resistant bacteria Methicillin-resistant Staphylococcus aureus have excellent inhibitory effect, and the bacteriostatic rate reaches more than 97%. The AIE molecule in the AIE composite electrospinning fiber membrane provided by Comparative Example 2 is tetra-(4-bromobenzene) ethylene, which is not the preferred tetra-styrene, 1,1,2,2-tetra-[4-carboxyl group in the present invention -(1,1-biphenyl)]ethylene or tetra-(4-hydroxyphenyl)ethylene, so it has little bacteriostatic effect. It can be seen that the selection of specific AIE molecules of the present invention endows the AIE composite electrospinning fiber membrane with efficient antibacterial properties, and beyond the preferred range of the present invention, antibacterial materials with good properties cannot be obtained.

Test Example 6

This test example is a bacterial morphology test experiment on the AIE composite electrospinning fiber membrane. The specific methods are as follows:

According to the method of step (1) in Test Example 5, Staphylococcus aureus was inoculated on the AIE composite electrospinning fiber membrane provided in Example 1, and after culturing in an incubator at 37°C for 24 hours, 2.5% glutaraldehyde and 30% glutaraldehyde were used. %, 50%, 70%, 80%, 90%, 95% and 100% ethanol dehydration, and the morphology of Staphylococcus aureus on the surface of the AIE composite electrospinning fiber membrane was observed with a scanning electron microscope, and the results in Example 1 were obtained. The morphology test chart of Staphylococcus aureus on the AIE composite electrospinning fiber membrane is shown in Figure 9. It can be seen from Figure 9 that the AIE molecules in the AIE composite electrospinning fiber membrane are released and wrapped in golden grapes. The surface of cocci, thereby destroying the bacterial cell wall, kills bacteria and inhibits proliferation.

The applicant declares that the present invention illustrates an AIE composite electrospinning fiber membrane of the present invention and its preparation method and application through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned process steps, that is, it does not mean that the present invention must rely on The above process steps can be implemented. Those skilled in the art should understand that any improvement to the present invention, the equivalent replacement of the selected raw materials of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims (10)

1. An AIE composite electrospinning fiber membrane, characterized in that, the AIE composite electrospinning fiber membrane is an electrospinning fiber membrane formed by a biocompatible polymer and AIE molecules. 2. The AIE composite electrospinning fiber membrane according to claim 1, wherein the AIE molecule is selected from the group consisting of tetrastyrene, 1,1,2,2-tetra-[4-carboxy-(1,1 -biphenyl)]ethylene or tetra-(4-hydroxyphenyl)ethylene or any one or a combination of at least two; Preferably, the biocompatible polymer is selected from polylactic acid, polylactic acid-glycolic acid copolymer, polycaprolactone, lactide-caprolactone copolymer, sodium alginate, chitosan or hyaluronic acid any one or a combination of at least two. 3. The AIE composite electrospinning fiber membrane according to claim 1 or 2, wherein the mass ratio of the biocompatible polymer to the AIE molecule is 1:(0.001～0.1), preferably 1: (0.01～0.08). 4 . The AIE composite electrospinning fiber membrane according to claim 1 , wherein the fiber diameter of the AIE composite electrospinning fiber membrane is 100-800 nm. 5 . 5. A preparation method of the AIE composite electrospinning fiber membrane according to any one of claims 1 to 4, wherein the preparation method comprises the following steps: (1) AIE molecule is mixed with organic solvent to obtain AIE solution; (2) mixing the biocompatible polymer with an organic solvent to obtain an electrospinning precursor; (3) mixing and dispersing the AIE solution obtained in step (1) and the electrospinning precursor solution obtained in step (2) to obtain an electrospinning solution; (4) Electrospinning the electrospinning solution obtained in step (3) to obtain the AIE composite electrospinning fiber membrane. 6. preparation method according to claim 5 is characterized in that, the organic solvent described in step (1) is selected from any one in dimethyl sulfoxide, N,N-dimethylformamide or acetone or at least a combination of the two; Preferably, the concentration of AIE molecules in the AIE solution of step (1) is 0.5-100 mg/mL; Preferably, the mixing described in step (1) is carried out under ultrasonic conditions; Preferably, the frequency of the ultrasound is 20-50 kHz; Preferably, the power of the ultrasound is 100-200W; Preferably, the ultrasonic time is 5～60min; Preferably, the AIE solution in step (1) is stored in the dark. 7. preparation method according to claim 5 or 6 is characterized in that, the organic solvent described in step (2) is selected from acetone, N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, Any one or a combination of at least two of chloroform, absolute ethanol, tetrahydrofuran or hexafluoroisopropanol; Preferably, the dispersion described in step (3) is carried out under dark conditions; Preferably, the dispersion time in step (3) is 2-12h; Preferably, the mass percentage of the biocompatible polymer in the electrospinning solution in step (3) is 10-30%; Preferably, the mass ratio of AIE molecules to the biocompatible polymer in the electrospinning solution in step (3) is (0.001-0.1):1. 8. The preparation method according to any one of claims 5 to 7, wherein the parameters of the electrospinning in step (4) are set as: the bolus injection rate of the electrospinning solution is 0.2-2 mL/h, The loading voltage is 10-20kV, the receiving distance is 10-25cm, and the diameter of the spinning nozzle is 0.3-1mm; Preferably, the preparation method specifically comprises the following steps: (1) Dissolving AIE molecules in an organic solvent under the assistance of ultrasound to obtain an AIE solution with an AIE molecular concentration of 0.5-100 mg/mL, and the AIE solution is protected from light; (2) dissolving the biocompatible polymer in an organic solvent to obtain an electrospinning precursor solution with a static mass percentage of the biocompatible polymer of 10-30%; (3) Mixing the AIE solution obtained in step (1) with the electrospinning precursor solution obtained in step (2), avoiding light and dispersing evenly, to obtain a mass ratio of AIE molecule to biocompatible polymer (0.001～0.1): 1 of the electrospinning solution; (4) Electrospinning the electrospinning solution obtained in step (3), the parameters of the electrospinning are set as: the injection speed of the electrospinning solution is 0.2-2 mL/h, and the loading voltage is 10-20 kV , the receiving distance is 10-25 cm, the diameter of the spinning nozzle is 0.3-1 mm, and the AIE composite electrospinning fiber membrane is obtained. 9 . The application of the AIE composite electrospinning fiber membrane according to any one of claims 1 to 4 in the preparation of biomedical materials. 10 . 10 . An antibacterial dressing, characterized in that, the antibacterial dressing comprises the AIE composite electrospinning fiber film according to any one of claims 1 to 4 .

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