CN115364270A

CN115364270A - Preparation method of antibacterial and antioxidant fiber film dressing containing traditional Chinese medicines

Info

Publication number: CN115364270A
Application number: CN202210372762.9A
Authority: CN
Inventors: 刘勇; 吕亚榕
Original assignee: Beijing University of Chemical Technology
Current assignee: Beijing University of Chemical Technology
Priority date: 2022-04-11
Filing date: 2022-04-11
Publication date: 2022-11-22

Abstract

The invention discloses a preparation method of a fiber film dressing which simultaneously contains traditional Chinese medicines of curcumin and borneol and has antibacterial and antioxidant properties, belonging to the technical field of traditional Chinese medicine preparations. According to the fiber film dressing prepared by the invention, active drugs curcumin and borneol are highly dispersed in a mixture of a carrier material gelatin and polycaprolactone, so that the dissolution rate of the drugs is effectively improved, the fiber film dressing has the effects of inhibiting staphylococcus aureus and the antioxidation characteristic, and the synergistic effect is shown after the drugs are combined. The three-dimensional porous structure with high specific surface area and high porosity can well simulate the structure of extracellular matrix, and is beneficial to realizing better healing of wounds. Can be used as an ideal and efficient wound dressing and has great application potential to wound surface wounds. Meanwhile, the preparation method adopts a solution electrostatic spinning method, has simple process, is green and efficient, has lower cost and is suitable for large-scale production.

Description

Preparation method of antibacterial and antioxidant fiber film dressing containing traditional Chinese medicines

Technical Field

The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a preparation method of a fibrous membrane which simultaneously contains traditional Chinese medicines of curcumin and borneol and has the effects of inhibiting staphylococcus aureus and oxidation resistance.

Background

With the continuous development of material science and medicine, the traditional wound dressing can not meet the requirements of patients, and the medical dressing with special functions is applied. For biomedical wound dressings, the use of materials with antimicrobial properties would be beneficial for preventing bacterial infections, sepsis, and the like. In addition, oxidative stress is one of the factors that causes the wound healing process to be slow, causing the wound to become a chronic disease. The destruction of Reactive Oxygen Species (ROS) at the wound site is considered an important strategy to accelerate the chronic wound healing process. Therefore, drugs, growth factors and the like are added into the wound dressing as bioactive additives, so that the wound can be healed more quickly and better. Curcumin (Cur) is a yellow phenolic substance extracted from the rhizome of turmeric, is one of the main active ingredients of turmeric, and has low toxicity, antibacterial, antioxidant and anti-inflammatory properties. Borneol (Borneol), belonging to bicyclic monoterpene compounds, has antibacterial and antiinflammatory effects. Meanwhile, the transdermal absorption of insoluble drugs can be promoted, and the blood concentration and bioavailability of other drugs can be improved. However, curcumin and borneol are oil-soluble and are insoluble in water, so that the bioavailability is low, and the clinical application of curcumin and borneol is greatly limited.

The electrostatic spinning nanofiber membrane can effectively improve the dissolution rate of insoluble drugs due to the characteristics of high specific surface area and high porosity, and the structural characteristics of the electrostatic spinning nanofiber membrane are similar to those of human natural tissues, so that the structure of extracellular matrix (ECM) can be well simulated. Among the many spinnable polymers, the natural polymer gelatin (Gel) is widely used in the biomedical field due to its unique biodegradation and biocompatibility. In addition, the water absorption capacity of the gelatin is 5 to 10 times of the self weight of the gelatin, and the gelatin has good moisture absorption performance. More importantly, the gelatin contains a large amount of glycine, proline and hydroxyproline, and can accelerate wound healing. But it has poor mechanical properties. The synthetic polymer Polycaprolactone (PCL) has excellent mechanical property, biocompatibility and biodegradability, and can make up for the defect of poor mechanical property of gelatin.

Disclosure of Invention

The invention aims to design a bacteriostatic antioxidant fiber film dressing containing traditional Chinese medicines so as to improve the dissolution performance of medicines in an aqueous medium and enable the dressing to have bacteriostatic and antioxidant effects.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the polymer is selected from gelatin (Gel) and Polycaprolactone (PCL), the medicine is curcumin (Cur) and Borneol (Borneol), and the solvent is Hexafluoroisopropanol (HFIP). Firstly, dissolving Gel and PCL in HFIP, then adding Cur with different masses into the solution according to the mass fractions of the Cur being 0%,1%,3%,5% and 3% in sequence, adding Borneol with different masses into the solution according to the mass fractions of Borneol being 1%,1%,1%,1% and 0% in sequence, placing the solution on a magnetic stirrer, stirring for 24 hours in the dark, and uniformly mixing to obtain the Gel/PCL/Cur/Borneol spinning solution with different concentration gradients.

The preparation method of the bacteriostatic antioxidant fiber membrane dressing containing traditional Chinese medicine is a solution electrostatic spinning method, the prepared spinning solution is filled into a 10ml medical injector, the injector is provided with a No. 21 steel needle, and the injector is fixed on a propulsion pump. The needle head is connected with a positive-pressure direct-current power supply, and the flat plate receiving device coated with the silicone oil paper is connected with a negative-pressure direct-current power supply. Starting an injection pump to push the spinning solution to advance, stretching the spinning solution under the action of an electric field force, volatilizing the solvent, solidifying the solute into filaments, and collecting the nanofiber membrane on a flat receiver. Wherein the parameters of the electrostatic spinning process are as follows: the positive pressure is 12kV, the negative pressure is 5kV, the advancing speed is 0.6ml/h, the receiving distance is 20cm, the environmental temperature is 20 ℃, the environmental humidity is 10% -30%, and the whole spinning process is protected from light.

After spinning, the fiber membrane and the silicone oil paper are placed in a constant-temperature drying oven together, dried for 24 hours at 50 ℃ to remove a solvent Hexafluoroisopropanol (HFIP) in the fiber membrane, and packaged in a sealed bag for storage.

The invention provides a preparation method of a traditional Chinese medicine-containing bacteriostatic antioxidant fibrous membrane dressing. The fiber diameter in the prepared fiber membrane is 0.5-0.7 μm, the fiber membrane has high specific surface area, high porosity and three-dimensional porous structure, and is beneficial to improving the contact area of curcumin and borneol with water, so that the dissolution rate of the medicine is improved, and the fiber membrane has excellent effects in bacteriostasis and antioxidation tests.

Compared with the prior art, the invention has the following advantages and effects

1. According to the fiber film dressing prepared by the invention, active drugs curcumin (Cur) and Borneol (Borneol) are highly dispersed in carrier materials of gelatin (Gel) and Polycaprolactone (PCL), so that the dissolution rate of the drug Cur is effectively improved, and the antibacterial property and the antioxidant property of the drug are improved.

2. The curcumin (Cur) and Borneol (Borneol) carried by the fiber film dressing prepared by the invention have antibacterial and antioxidant effects, and show synergistic effect after the drugs are combined.

3. The fiber membrane dressing provided by the invention has the advantages of high specific surface area, high porosity and three-dimensional porous structure, can well simulate the structure of extracellular matrix (ECM), and is beneficial to realizing better healing of wounds.

4. The fiber film dressing of the invention uses carrier materials of gelatin (Gel) and Polycaprolactone (PCL) which are biodegradable, and has very good practical prospect.

5. The fiber film dressing is prepared by using a solution electrostatic spinning method, and the preparation method is simple, low in cost, green, high-efficiency, easy to operate and suitable for large-scale production.

Drawings

FIG. 1 is SEM images of fiber film dressings prepared in examples 1, 2, 3 and 4 of the present invention

FIG. 2 is a graph showing in vitro dissolution profiles of the fiber film dressings prepared in examples 2, 3 and 4 of the present invention

FIG. 3 is a bacteriostatic diagram of the fiber film dressing prepared in example 3 of the present invention

FIG. 4 is a bar graph of the antioxidant performance of the fiber film dressings prepared in examples 3 and 5 of the present invention

Detailed Description

In order to explain the bacteriostatic and antioxidant fiber membrane dressing containing traditional Chinese medicine and the preparation method thereof more fully, the following specific examples of the preparation method are provided, but the invention is not limited to the examples.

Example 1:

0.35g of gelatin (Gel), 0.15g of Polycaprolactone (PCL) and 0.05g of Borneol (Borneol) are weighed out and dissolved in 5ml of Hexafluoroisopropanol (HFIP), and the obtained solution is placed on a magnetic stirrer and stirred for 24 hours to form a uniform spinning solution.

The prepared spinning solution was loaded into a 10ml medical syringe and fixed to a propeller pump using a solution electrospinning method. The flat plate receiving device coated with the silicone oil paper is connected with a positive voltage of 12kV, the needle head is connected with a negative voltage of 5kV, the distance between the needle head and the flat plate receiving device is 20cm, the environmental temperature is 20 ℃, and the environmental humidity is 10% -30%. Starting an injection pump to push the spinning solution at the advancing speed of 0.6ml/h, stretching the spinning solution under the action of an electric field force, volatilizing the solvent, solidifying the solute into filaments, and collecting the nanofiber membrane on a flat receiver.

Placing the fiber membrane and the silicone oil paper together in a constant temperature drying oven, drying at 50 deg.C for 24h to remove solvent Hexafluoroisopropanol (HFIP) in the fiber membrane, and packaging the fiber membrane in a sealed bag for storage.

Example 2:

0.35g of gelatin (Gel), 0.15g of Polycaprolactone (PCL) and 0.05g of Borneol (Borneol) are weighed and dissolved in 5ml of Hexafluoroisopropanol (HFIP), and then 0.05g of curcumin (Cur) is added to place the obtained solution on a magnetic stirrer and stir in the dark for 24h to form a uniform spinning solution.

(2) The prepared spinning solution was loaded into a 10ml medical syringe fixed to a propeller pump using a solution electrospinning method. The flat plate receiving device coated with the silicone oil paper is connected with positive pressure of 12kV, the needle head is connected with negative pressure of 5kV, the distance between the needle head and the flat plate receiving device is 20cm, the environmental temperature is 20 ℃, and the environmental humidity is 10% -30%. Starting an injection pump to push the spinning solution at the advancing speed of 0.6ml/h, stretching the spinning solution under the action of electric field force, volatilizing the solvent, solidifying the solute into filaments, collecting the nanofiber membrane on a flat receiver, and keeping the spinning process away from light as far as possible.

(3) The fiber membrane and the silicon oil paper are placed in a constant temperature drying oven together, dried for 24 hours at 50 ℃ to remove the solvent Hexafluoroisopropanol (HFIP) in the fiber membrane, and then the fiber membrane is sealed in a sealed bag for storage.

Example 3:

0.35g of gelatin (Gel), 0.15g of Polycaprolactone (PCL) and 0.05g of Borneol (Borneol) are weighed and dissolved in 5ml of Hexafluoroisopropanol (HFIP), and then 0.15g of curcumin (Cur) is added to place the obtained solution on a magnetic stirrer and stir in the dark for 24h to form a uniform spinning solution.

(2) The procedure was as in example 2.

Example 4:

0.35g of gelatin (Gel), 0.15g of Polycaprolactone (PCL) and 0.05g of Borneol (Borneol) are weighed out and dissolved in 5ml of Hexafluoroisopropanol (HFIP), and then 0.25g of curcumin (Cur) is added to stir the obtained solution on a magnetic stirrer for 24 hours in the dark to form a uniform spinning solution.

The procedure was as in example 2.

Example 5:

0.35g of gelatin (Gel) and 0.15g of Polycaprolactone (PCL) are weighed out and dissolved in 5ml of Hexafluoroisopropanol (HFIP), 0.15g of curcumin (Cur) is subsequently added and the solution is stirred on a magnetic stirrer for 24h in the dark to form a homogeneous spinning solution.

(2) The procedure was as in example 2.

SEM test characterization is carried out on the fiber film dressings prepared in the above examples 1, 2, 3 and 4, and the results show that the fibers are good in appearance and uniform in distribution. The magnified detail shows that the fiber surface is smooth and no obvious particles are present. As shown in fig. 1. With the increase of the content of curcumin (Cur), the conductivity of the solution is increased, and the solution is subjected to larger stretching force in an electric field, so that the diameter is thinned, and the diameter distribution range is 0.5-0.7 μm.

The fiber film dressings prepared in the above examples 2, 3 and 4 were subjected to in vitro dissolution measurement: respectively taking 0.005g, 0.0167g and 0.01g of the fiber membranes, immersing the fiber membranes into 100mL of PBS (containing 0.4% of Tween-80), putting the fiber membranes into a constant temperature oscillator, wherein the rotating speed of the constant temperature oscillator is 100r/min, the medium temperature is 37 ℃, sampling 4mL at regular intervals, simultaneously adding PBS (phosphate buffer solution) with the same volume and temperature, centrifuging the obtained solution at a high speed of 5000r/min, taking supernate, and measuring the absorbance of the supernate at the wavelength of 427nm by using an ultraviolet spectrophotometer to calculate the dissolution rate of curcumin (Cur). The dissolution curves shown in figure 2 are obtained, and the drug release rates of the fibrous membrane reach 66.02%,80.74% and 74.19% respectively.

The fiber film dressing prepared in the above example 3 was tested for antibacterial performance, and the fiber film obtained in the above example 3 of the present invention was selected to study the in vitro antibacterial activity against staphylococcus aureus: 100 μ L of Staphylococcus aureus suspension with a concentration of 1X 106cfu/mL was inoculated into sterilized agar, and the fibrous membrane disc obtained in example 3 above, containing 50 μ g of curcumin (Cur) by mass, was cut out, and curcumin (Cur) and Borneol (Borneo) with the same mass were dropped into the same size of disc as a control group. Sterilizing with ultraviolet rays for 30min, placing into the agar, culturing at 37 deg.C for 24 hr, and observing antibacterial effect. As shown in FIG. 3, the fibrous film dressing prepared in example 3 of the present invention has a significant inhibitory effect on Staphylococcus aureus. In addition, for the borneol-free curcumin film (C) and the borneol fiber film (B), no antibacterial activity was exhibited, which is suspected to be related to the drug concentration, and it was also demonstrated that the two had stronger antibacterial activity after combined use.

The fiber film dressings containing Borneol (Borneol) and not containing Borneol (Borneol) prepared in the above examples 3 and 5 were subjected to an antioxidant performance test, and the fiber films obtained in the above examples 3 and 5 of the present invention were selected to study the antioxidant activity: a certain amount of the above fiber membrane was immersed in 100mL of methanol: distilled water =1:1, placing the mixed solvent in a constant-temperature oscillator at the rotation speed of 100r/min and the medium temperature of 37 ℃, oscillating at the constant temperature for 24 hours to obtain a dissolution liquid, respectively taking 0.5ml, 0.75ml, 1.0ml, 1.25ml and 1.5ml of dissolution liquid in a centrifugal tube, and adding methanol: distilled water =1: the mixed solvent of 1 was made to a constant volume of 2ml to obtain curcumin concentrations of 50ppm, 75ppm, 100ppm, 125ppm and 150ppm in this order in the mixed solution, and then 3ml of 1, 1-diphenyl-2-picrylhydrazino (DPPH) solution was added, respectively. Wait for 60min. And (3) respectively measuring the absorbance of the DPPH solution and the mixed solution by using an ultraviolet spectrophotometer at the wavelength of 517nm, calculating the antioxidant efficiency of the DPPH solution and the mixed solution, and determining the optimal antioxidant drug concentration to be 125ppm. The change of the antioxidant performance of the medicine with time is measured under the concentration, and the antioxidant efficiency of the bacteriostatic antioxidant fiber membrane dressing containing Borneol (Borneol) and the antioxidant efficiency of the bacteriostatic antioxidant fiber membrane dressing not containing Borneol (Borneol) reach the maximum of 81 percent and 85 percent respectively within 180 min. Shows better antioxidant activity. In addition, the antioxidant capacity of the bornyl alcohol (Borneol) -containing fiber film dressing is higher than that of the bornyl alcohol (Borneol) -free fiber film dressing, which shows that the antioxidant capacity of the combined medicament is stronger than that of a single medicament.

Claims

1. A preparation method of a traditional Chinese medicine-containing antibacterial and antioxidant fiber film dressing is characterized by comprising the following specific steps:

(1) The polymer is selected from gelatin (Gel) and Polycaprolactone (PCL), the medicine is curcumin (Cur) and Borneol (Borneol), and the solvent is Hexafluoroisopropanol (HFIP); firstly, dissolving Gel and PCL in HFIP, then adding Cur with different masses into the solution according to the mass fractions of the Cur being 0%,1%,3%,5% and 3% in sequence, adding Borneol with different masses into the solution according to the mass fractions of Borneol being 1%,1%,1%,1% and 0% in sequence, placing the solution on a magnetic stirrer, stirring for 24 hours in the dark, and uniformly mixing to prepare Gel/PCL/Cur/Borneol spinning solutions with different concentration gradients;

(2) Filling the prepared spinning solution into a 10ml medical injector, wherein the injector is provided with a 21# steel needle head, and fixing the injector on a propulsion pump; the needle head is connected with a positive-pressure direct-current power supply, and the flat plate receiving device coated with the silicone oil paper is connected with a negative-pressure direct-current power supply; starting an injection pump to push the spinning solution to advance, stretching the spinning solution under the action of an electric field force, volatilizing a solvent, solidifying a solute into filaments, and collecting a nanofiber membrane on a flat receiver; wherein the electrostatic spinning parameters are as follows: the positive pressure is 12kV, the negative pressure is 5kV, the propelling speed is 0.6ml/h, the receiving distance is 20cm, the environmental temperature is 20 ℃, the environmental humidity is 10-30%, and the whole electrostatic spinning process is protected from light;

(3) After spinning, the fiber membrane and the silicone oil paper are placed in a constant-temperature drying oven together, dried for 24 hours at 50 ℃ to remove a solvent Hexafluoroisopropanol (HFIP) in the fiber membrane, and packaged in a sealed bag for storage.