CN106267309B

CN106267309B - alginate composite antibacterial dressing and preparation method thereof

Info

Publication number: CN106267309B
Application number: CN201610692139.6A
Authority: CN
Inventors: 卢亢; 陈锦涛; 熊亮
Original assignee: Guangdong Taibao Medical Science Technology Co Ltd
Current assignee: Guangdong Taibao Medical Science Technology Co Ltd
Priority date: 2016-08-19
Filing date: 2016-08-19
Publication date: 2019-12-10
Anticipated expiration: 2036-08-19
Also published as: CN106267309A

Abstract

the invention belongs to the field of medical biological high polymer materials, and particularly relates to an alginate composite antibacterial dressing and a preparation method thereof. The silk fibroin composite membrane is used as an antibacterial agent, a humectant and an anti-adhesion agent, and is loaded on alginate fibers, and the finally processed alginate composite antibacterial dressing can play a role in protecting wound surfaces, inhibiting bacteria, stopping bleeding, preventing adhesion and promoting wound healing clinically. The alginate composite antibacterial dressing has the advantages of improving the mechanical property of alginate in a wet state, overcoming the adhesion of alginate in a wound with blood seepage, endowing alginate with a certain antibacterial function, being suitable for nursing a typical wound surface wound, accelerating the healing of the wound surface wound and relieving the pain of a wound patient, along with simple process, easy processing and realization of mass production.

Description

Alginate composite antibacterial dressing and preparation method thereof

Technical Field

The invention belongs to the field of medical biological high polymer materials, and particularly relates to an alginate composite antibacterial dressing and a preparation method thereof. The silk fibroin composite membrane is used as an antibacterial agent, a humectant and an anti-adhesion agent, and is loaded on alginate fibers, and the finally processed alginate composite antibacterial dressing can play a role in protecting wound surfaces, inhibiting bacteria, stopping bleeding, preventing adhesion and promoting wound healing clinically.

Background

Skin wounds are skin defects caused by mechanical, electrical, thermal, chemical, etc. factors or due to physiological allergies. Due to the prevalence of human skin wounds, a wide variety of wound care methods have been summarized over the last millennium. Since 1962, Winten by animal physiologist discovered through research that rapid wound healing was more favored in a wet environment. After that, scientists have continuously verified the accuracy of this conclusion. After the 70 s in the 20 th century, various novel dressings based on the 'moist healing theory of wounds' have been produced, and alginate has occupied an important place in the ranks of novel dressings due to the advantages of hemostasis, imbibition, moisture retention, no skin irritation, good biocompatibility and the like. The prior documents and patents have a great deal of concern about the application of alginate in medical wound dressings, but the wide application of alginate in clinic is limited due to the defect of the bacteriostatic function of the alginate. In addition, the secondary injury of the wound caused by the adhesion of the alginate dressing in the wound with more bleeding is always a problem that the academia continuously seeks to solve by an improved scheme.

Silk fibroin is a natural biological protein, has excellent biocompatibility and good mechanical and physical properties, and attracts more and more attention and researches in the field of biological materials in recent years. The silk fibroin can be widely applied to biological materials such as artificial ligaments, artificial bones, artificial blood and the like, and can be used as a good drug slow-release material. In the aspect of nursing skin wounds, the excellent cell biology promotion property and the low inflammatory reactivity of the composition also make the composition have wide application prospects. Since the degummed silk fibroin has good film forming property and is easy to form a composite film with other functional materials, the silk fibroin film is a popular research topic. The application of the composite material formed by silk fibroin and alginate in dressing is reported, however, the formed composite material mostly has a new function of adding materials, and reports that the adhesion prevention effect and the mechanical strength of the composite material formed by the silk fibroin and the alginate are improved are not discovered for a while.

Disclosure of Invention

The invention aims to overcome the defects of alginate functional dressings, overcome the defects in the prior art and provide an alginate composite antibacterial dressing and a preparation method thereof. The preparation method has simple process, is easy to process and realize batch production, and the prepared alginate composite antibacterial dressing improves the mechanical property of alginate in a wet state, overcomes the adhesion of alginate in a wound with blood seepage, endows alginate with a certain antibacterial function, is suitable for nursing a typical wound surface wound, accelerates the healing of the wound surface wound and relieves the pain of a wound patient.

In order to solve the technical problems, the invention adopts the technical scheme that:

an alginate composite antibacterial dressing is composed of a silk fibroin composite membrane and alginate fibers.

Further, the content of the silk fibroin composite membrane in the antibacterial dressing is 30 ~ 150 mg/g.

Furthermore, the aperture of the silk fibroin composite membrane is 20 ~ 150 μm.

Preferably, the thickness of the silk fibroin composite membrane in the antibacterial dressing is 1 ~ 30 μm.

A preparation method of alginate composite antibacterial dressing comprises the following specific steps:

S1, preparing alginate fibers, namely pre-dissolving alginate, stirring, filtering by using a filter cloth under the pressure of 0.1 ~ 0.3.3 MPa to obtain alginate spinning solution, drawing, washing, dividing filaments and winding by using a wet spinning process and taking a calcium ion aqueous solution or an alcohol solution with the mass fraction of 5% of ~ 10% as a coagulating bath, and drying at the temperature of 100 ℃ in hot air to obtain the alginate fibers;

S2, preparing a silk fibroin solution, namely soaking natural silkworm shells or silk in a mixed organic solvent in sequence by adopting a salt-dissolving enzymolysis process, boiling the mixture in water for degumming, dissolving a mixed solution consisting of inorganic salt and formic acid, dialyzing, filtering and concentrating to obtain the silk fibroin solution with the mass concentration of 15 ~ 50%, wherein the mixed organic solvent is used for soaking to remove wax substances, carbohydrates and ash impurities in the natural silkworm shells or silk;

S3, preparing a silk fibroin film, namely sequentially adding an organosilicon quaternary ammonium salt solution with the mass concentration of 12 ~ 20%, a sugar solution with the mass concentration of 1 ~ 10%, a hyaluronic acid solution with the mass concentration of 2 ~ 6%, a vitamin E solution with the mass concentration of 0.5 ~ 1% and a hyaluronic acid solution with the mass concentration of 0.1 ~ 0.5.5% into the silk fibroin solution prepared in the step S2, fully stirring and mixing for 0.5 ~ 2h, standing and defoaming to obtain a silk fibroin film mixed solution, adopting organosilicon quaternary ammonium salt to enable the formed silk fibroin film to generate an electrostatic effect and interfere with the synthesis of bacterial cell walls so as to generate a good antibacterial effect, and meanwhile, utilizing the flexibility of organosilicon to enable the composite film formed by the organosilicon quaternary ammonium salt and the silk fibroin to improve the mechanical property and the anti-adhesion property of the silk fibroin film;

S4, loading a silk fibroin film, namely adding 1 ~ 20% of dihydric alcohol into the silk fibroin film mixed solution prepared in the step S3, wherein the volume ratio of the silk fibroin film mixed solution to ethylene glycol is 100: 1 ~ 10 to form a mixed solution, padding the alginate fibers prepared in the step S1 into the mixed solution under the conditions of constant temperature and constant humidity for 20 ~ 60min, taking out the alginate fibers, pre-freezing the alginate fibers at the temperature of-80 ~ -20 ℃ for 10 ~ 24h, and finally performing vacuum drying for 18 ~ 24h under the condition of low temperature to obtain the membrane-loaded alginate composite fiber, wherein the silk fibroin composite film is of a porous structure and is loaded on the surface of the alginate fibers, the functional characteristics of the alginate fibers are not influenced, the functions of other functional components in the composite film can be fully exerted, all the functional components generate a synergistic effect, and the wound healing time is greatly shortened;

S5, preparation of alginate composite antibacterial dressing: and (5) opening, carding, lapping, reinforcing, drying and coiling the film-carrying alginate composite fiber prepared in the step S4, and finally packaging and sterilizing to obtain the alginate composite antibacterial dressing.

Further, in the step S1, the pre-dissolving temperature is 30 ℃ ~ 40 ℃, the time is 2h ~ 3h, the stirring temperature is 30 ℃ ~ 60 ℃, the stirring speed is 200r/min ~ 400r/min, the stirring time is 3h ~ 7h, the temperature of the wet spinning process is 60 ~ 80 ℃, the number of spinneret holes is 200 ~ 500, and the spinning speed is 5 ~ 100 m/min.

furthermore, in the step S2, the mixed organic solvent is formed by mixing ethers and alcohols according to a volume ratio of 1:1, the ethers are methyl ether or ethyl ether, the alcohols are methanol or ethanol, the inorganic salt and formic acid according to a volume ratio of 1:1 form a mixed solution, the mass concentration of the inorganic salt is 1 ~ 20%, the mass concentration of the formic acid is 50 ~ 90%, and the inorganic salt is one of sodium chloride, potassium chloride, sodium hydrosulfide and potassium hydrosulfide.

preferably, in the step S3, the volume ratio of the silk fibroin solution, the organosilicon quaternary ammonium salt solution, the sugar solution, the hyaluronic acid solution, the vitamin E solution and the hyaluronic acid solution is 12 ~ 18: 2 ~ 6: 1 ~ 4: 4 ~ 8: 3 ~ 7: 2 ~ 5.

Preferably, the organosilicon quaternary ammonium salt solution is one of quaternized haloalkylsilane, quaternized amino silicone oil and quaternized alkyl silicone oil; the sugar solution is one or more of glucose, lactose, galactose, mannose, ribose, sorbose, gulose, fucose, rhamnose, sucrose, fructose, maltose and cellobiose.

preferably, the dihydric alcohol in the step S4 is one or more of ethylene glycol, 1, 3-propylene glycol, 2, 3-propylene glycol, 1, 4-butanediol, 1, 2-butanediol, 1, 3-butanediol, 2, 3-butanediol, diethylene glycol and triethylene glycol.

the invention has the beneficial effects that:

1) The alginate is a biological material with good biocompatibility, calcium ions in the alginate are easy to exchange and adsorb sodium ions in hemoglobin, the calcium ions can enter plasma to generate a blood coagulation effect, so that the dressing has excellent blood coagulation, and the alginate has good liquid absorption and moisture retention due to the characteristics of the molecular structure of the alginate, so that the healing of the product is facilitated;

2) The silk fibroin film contains natural and rich protein components, can promote cell biology, is beneficial to the biology of new granulation of wounds and is beneficial to promoting the healing of the wounds;

3) The organic silicon in the composite membrane contains amino, so that the composite membrane can generate an electrostatic effect and can interfere the synthesis of bacterial cell walls, thereby generating a better antibacterial effect;

4) By utilizing the flexibility of the organic silicon, the composite membrane formed by the organic silicon and the silk fibroin can improve the mechanical property and the anti-adhesion property of the membrane;

5) the silk fibroin composite membrane is a porous structure, the alginate functional characteristics are not influenced when the silk fibroin composite membrane is loaded on the surface of alginate fibers, the functions of other functional components in the composite membrane can be fully exerted, all functional component substances generate a synergistic effect, and the wound healing time is greatly shortened.

drawings

FIG. 1 is a graph showing the growth of blank 0-exposed bacteria in example 6 of the present invention;

FIG. 2 is a graph of 18h exposed bacterial growth for a blank in example 6 of the present invention;

FIG. 3 is a graph showing the growth of 0h exposed bacteria in the experimental group of example 6 of the present invention;

FIG. 4 is a graph showing the growth of 18h exposed bacteria in the experimental group of example 6 of the present invention;

FIG. 5 is a graph showing the growth of 0h exposed bacteria in the control group in example 6 of the present invention;

FIG. 6 is a graph showing the growth of 18h exposed bacteria in the control group in example 6 of the present invention.

Detailed Description

example 1

(1) preparation of alginate fibers: soaking alginate at 30 ℃ for 2.5h for pre-dissolving, then stirring for 4h under the conditions of 60 ℃ and 200r/min of rotating speed or stirring for 4h under the conditions of 30 ℃ and 200r/min of rotating speed, and finally filtering by adopting filter cloth under the pressure of 0.1MPa to obtain alginate spinning solution; spinning the spinning solution at 60 ℃ by using spinning holes with the number of 300, wherein the spinning speed is 5m/min, then solidifying in a solidification bath containing 5 mass percent of calcium ion water solution or alcohol solution, stretching, washing, dividing filaments, winding, and drying in hot air at 100 ℃ to obtain alginate fibers;

(2) Preparation of the silk fibroin film: preparing a silk fibroin solution: adopting a salt-dissolving enzymolysis process, sequentially treating natural silkworm shells or silks with methyl ether and methanol according to a volume ratio of 1:1, soaking in an organic solvent formed by mixing to remove waxy substances, carbohydrates and ash impurities, then degumming by boiling in water, and mixing sodium chloride solution with the mass concentration of 20wt% and formic acid solution with the mass concentration of 50wt% according to the volume ratio of 1:1, then dialyzing, filtering and concentrating to prepare a silk fibroin solution with the mass concentration of 15%; preparing a silk fibroin film: adding a quaternized haloalkyl silane solution with the mass concentration of 12%, a glucose solution with the mass concentration of 10%, a hyaluronic acid solution with the mass concentration of 5%, a vitamin E solution with the mass concentration of 0.8% and a hyaluronic acid solution with the mass concentration of 0.3% in sequence into the silk fibroin solution, wherein the weight percentage of the added components in the silk fibroin solution is as follows: organic silicon quaternary ammonium salt solution: glucose solution: hyaluronic acid solution: vitamin E solution: the volume ratio of the hyaluronic acid solution is 12: 6: 1: 8: 3: 2, fully stirring and mixing the mixed solution for 0.6h, standing and defoaming to obtain a silk fibroin membrane mixed solution;

(3) Loading the silk fibroin film: adding 20% by mass of ethylene glycol into the silk fibroin film mixed solution, wherein the volume ratio of the silk fibroin film mixed solution to the ethylene glycol is 100: 1, forming a mixed solution, padding the alginate fibers prepared in the step (1) into the mixed solution for 20min under the conditions of constant temperature and constant humidity, taking out the alginate fibers, pre-freezing the alginate fibers at the low temperature of-80 ℃, pre-freezing for 10h, and finally vacuum-drying for 24h under the low temperature condition to obtain the membrane-carrying alginate composite fibers;

(4) preparation of alginate composite antibacterial dressing: and (4) opening, carding, lapping, reinforcing, drying and coiling the film-carrying alginate composite fiber prepared in the step (3), and finally packaging and sterilizing to obtain the alginate composite antibacterial dressing.

The alginate composite antibacterial dressing prepared by the embodiment is composed of a silk fibroin composite membrane and alginate fibers, wherein the content of the silk fibroin composite membrane is 150mg/g, the pore diameter of the silk fibroin composite membrane is 20 micrometers, and the thickness of the silk fibroin composite membrane is 15 micrometers.

Example 2

(1) preparation of alginate fibers: soaking alginate at 40 ℃ for 2h for pre-dissolving, then stirring for 3h under the conditions of 40 ℃ and 400r/min of rotation speed or stirring for 6h under the conditions of 45 ℃ and 300r/min of rotation speed, and finally filtering by adopting filter cloth under the pressure of 0.3MPa to obtain alginate spinning solution; spinning the spinning solution at 80 ℃ by using spinning holes with the number of 200, wherein the spinning speed is 100m/min, then solidifying in a solidification bath containing 8 mass percent of calcium ion aqueous solution or alcohol solution, stretching, washing, dividing filaments, winding, and drying in hot air at 80 ℃ to obtain alginate fibers;

(2) preparation of the silk fibroin film: preparing a silk fibroin solution: adopting a salt-dissolving enzymolysis process, sequentially processing natural silkworm shells or silks by methyl ether and ethanol according to a volume ratio of 1:1, soaking in an organic solvent formed by mixing to remove waxy substances, carbohydrates and ash impurities, then degumming by boiling in water, and mixing sodium thiocyanate solution with the mass concentration of 1wt% and formic acid solution with the mass concentration of 90wt% according to the volume ratio of 1:1, then dialyzing, filtering and concentrating to prepare a silk fibroin solution with the mass concentration of 50%; preparing a silk fibroin film: adding a quaternized amino silicone oil solution with the mass concentration of 20%, a mannose solution with the mass concentration of 1%, a hyaluronic acid solution with the mass concentration of 6%, a vitamin E solution with the mass concentration of 0.5% and a hyaluronic acid solution with the mass concentration of 0.4% in sequence into the silk fibroin solution, wherein the weight percentage of the quaternized amino silicone oil solution to the silk fibroin solution is as follows: organic silicon quaternary ammonium salt solution: mannose solution: hyaluronic acid solution: vitamin E solution: the volume ratio of the hyaluronic acid solution is 18: 2: 4: 4: 7: 4, fully stirring and mixing the mixed solution for 0.5h, standing and defoaming to obtain a silk fibroin membrane mixed solution;

(3) Loading the silk fibroin film: adding 18% 2, 3-propylene glycol by mass concentration into the silk fibroin film mixed solution, wherein the volume ratio of the silk fibroin film mixed solution to the 2, 3-propylene glycol is 100: 6, forming a mixed solution, padding alginate fibers into the mixed solution for 40min under the conditions of constant temperature and constant humidity, taking out the alginate fibers, pre-freezing at the low temperature of-60 ℃ for 24h, and finally, drying in vacuum for 20h under the low temperature condition to obtain the film-carrying alginate composite fiber;

the alginate composite antibacterial dressing prepared by the embodiment is composed of a silk fibroin composite membrane and alginate fibers, wherein the content of the silk fibroin composite membrane is 30mg/g, the pore diameter of the silk fibroin composite membrane is 150 micrometers, and the thickness of the silk fibroin composite membrane is 30 micrometers.

Example 3

(1) Preparation of alginate fibers: soaking alginate at 35 ℃ for 3h for pre-dissolving, then stirring for 3.5h under the conditions of 50 ℃ and 300r/min of rotation speed or stirring for 7h under the conditions of 60 ℃ and 400r/min of rotation speed, and finally filtering by adopting filter cloth under the pressure of 0.2MPa to obtain alginate spinning solution; spinning the spinning solution at 70 ℃ by using spinning holes with the number of 500, wherein the spinning speed is 60m/min, then solidifying in a solidification bath containing 10 mass percent of calcium ion aqueous solution or alcohol solution, stretching, washing, dividing filaments, winding, and drying in hot air at 70 ℃ to obtain alginate fibers;

(2) preparation of the silk fibroin film: preparing a silk fibroin solution: adopting a salt-dissolving enzymolysis process, sequentially carrying out ethyl ether and methanol treatment on natural silkworm shells or silk according to a volume ratio of 1:1, soaking in an organic solvent formed by mixing to remove waxy substances, carbohydrates and ash impurities, then degumming by boiling in water, and mixing potassium thiocyanate solution with the mass concentration of 15wt% and formic acid solution with the mass concentration of 70wt% according to the volume ratio of 1:1, then dialyzing, filtering and concentrating to prepare a silk fibroin solution with the mass concentration of 45%; preparing a silk fibroin film: adding a quaternized alkyl silicone oil solution with the mass concentration of 15%, a fucose solution with the mass concentration of 5%, a hyaluronic acid solution with the mass concentration of 2%, a vitamin E solution with the mass concentration of 1% and a hyaluronic acid solution with the mass concentration of 0.1% in sequence into the silk fibroin solution, wherein the weight ratio of the quaternized alkyl silicone oil solution to the hyaluronic acid solution is as follows: organic silicon quaternary ammonium salt solution: fucose solution: hyaluronic acid solution: vitamin E solution: the volume ratio of the hyaluronic acid solution is 15: 4: 2: 5: 4: 3, fully stirring and mixing the mixed solution for 2 hours, standing and defoaming to obtain a silk fibroin membrane mixed solution;

(3) loading the silk fibroin film: adding 16 mass percent of 1, 4-butanediol into the silk fibroin film mixed solution, wherein the volume ratio of the silk fibroin film mixed solution to the 1, 4-butanediol is 100: 8, forming a mixed solution, padding alginate fibers into the mixed solution for 50min under the conditions of constant temperature and constant humidity, taking out the alginate fibers, pre-freezing at the low temperature of-20 ℃ for 18h, and finally, vacuum-drying at the low temperature for 22h to obtain the film-carrying alginate composite fiber;

the alginate composite antibacterial dressing prepared by the embodiment is composed of a silk fibroin composite membrane and alginate fibers, wherein the content of the silk fibroin composite membrane is 90mg/g, the pore diameter of the silk fibroin composite membrane is 100 micrometers, and the thickness of the silk fibroin composite membrane is 1 micrometer.

Example 4

(1) Preparation of alginate fibers: soaking alginate at 36 ℃ for 2 hours for pre-dissolving, then stirring for 3 hours at the temperature of 45 ℃ and the rotating speed of 350r/min or stirring for 5 hours at the temperature of 55 ℃ and the rotating speed of 350r/min, and finally filtering by adopting filter cloth under the pressure of 0.25MPa to obtain alginate spinning solution; spinning the spinning solution at 75 deg.c with 400 holes in spinning speed of 80m/min, coagulating in coagulating bath containing water solution or alcohol solution of calcium ion in 9 wt%, stretching, water washing, splitting, winding and drying in 90 deg.c hot air to obtain alginate fiber;

(2) preparation of the silk fibroin film: preparing a silk fibroin solution: adopting a salt-dissolving enzymolysis process, sequentially carrying out treatment on natural silkworm shells or silk by using diethyl ether and ethanol according to a volume ratio of 1:1, soaking in an organic solvent formed by mixing to remove waxy substances, carbohydrates and ash impurities, then degumming by boiling in water, and mixing potassium chloride solution with the mass concentration of 10wt% and formic acid solution with the mass concentration of 60wt% according to the volume ratio of 1:1, then dialyzing, filtering and concentrating to prepare a silk fibroin solution with the mass concentration of 48%; preparing a silk fibroin film: adding a quaternized haloalkyl silane solution with the mass concentration of 18%, a cellobiose solution with the mass concentration of 6%, a hyaluronic acid solution with the mass concentration of 4%, a vitamin E solution with the mass concentration of 0.6% and a hyaluronic acid solution with the mass concentration of 0.5% in sequence into the silk fibroin solution, wherein the weight ratio of the silk fibroin solution: organic silicon quaternary ammonium salt solution: cellobiose solution: hyaluronic acid solution: vitamin E solution: the volume ratio of the hyaluronic acid solution is 16: 5: 3: 7: 6: 5, fully stirring and mixing the mixed solution for 1h, standing and defoaming to obtain a silk fibroin membrane mixed solution;

(3) Loading the silk fibroin film: 1, 3-butanediol with the mass concentration of 1% is added into the silk fibroin film mixed solution, and the volume ratio of the silk fibroin film mixed solution to the 1, 3-butanediol is 100: 10, forming a mixed solution, padding alginate fibers into the mixed solution for 60min under the conditions of constant temperature and constant humidity, taking out the alginate fibers, pre-freezing at the low temperature of-40 ℃ for 20h, and finally, drying in vacuum for 18h under the low temperature condition to obtain the film-carrying alginate composite fiber;

The alginate composite antibacterial dressing prepared by the embodiment is composed of a silk fibroin composite membrane and alginate fibers, wherein the content of the silk fibroin composite membrane is 60mg/g, the pore diameter of the silk fibroin composite membrane is 120 micrometers, and the thickness of the silk fibroin composite membrane is 10 micrometers.

in example 1 ~ 4 of the present invention, examples of the sugar solution include glucose, mannose, fucose and cellobiose, examples of the dihydric alcohol include ethylene glycol, 2, 3-propanediol, 1, 4-butanediol and 1, 3-butanediol, examples of the present invention include, but are not limited to, lactose, galactose, ribose, sorbose, gulose, rhamnose, sucrose, fructose and maltose, and examples of the dihydric alcohol include one or more of 1, 3-propanediol, 1, 2-butanediol, 2, 3-butanediol, diethylene glycol and triethylene glycol.

Example 5

Experimental groups: the alginate composite antibacterial dressing prepared in example 1 is used as an experimental group;

control group: taking alginate dressing without loading silk fibroin film as a control group;

The mechanical property of the dressing prepared by processing the experimental group and the control group by the same method is tested, and the normal state is the mechanical property in a dry state; the wet state is the mechanical property after soaking in simulated human body exudate, and the result is shown in the following table:

TABLE 1 comparison of tensile Strength of dressings of Experimental group and control group (Unit: MPa)

Table 2 comparison of elongation at break (%)' of dressing of experimental group and control group

as can be seen from the comparison of the mechanical property data in tables 1 and 2, the alginate composite antibacterial dressing product prepared by the invention has obviously improved mechanical property and is more suitable for nursing wounds caused by liquid seepage.

example 6

experimental groups: the alginate composite antibacterial dressing prepared in example 2 was used as an experimental group;

The dressing prepared by the same method for the experimental group and the control group is subjected to antibacterial performance test, the test method comprises the steps of respectively shearing the dressings of the experimental group and the control group, washing to be neutral, respectively placing the dressings in triangular flasks, respectively adding 10ml of 1/10AATCC nutrient broth and 10ml of buffered physiological saline (pH =7.0 ~ 7.2.2), sterilizing in a high-pressure steam sterilizer, cooling to room temperature, inoculating Candida albicans strain, placing in a constant-temperature shaking table at 37 ℃ for oscillation, taking 0.1 ml of the solution after a certain time, spreading the solution on a nutrient agar culture medium, culturing in a constant-temperature box at 37 ℃ for 24h, and counting viable bacteria, wherein blank control tests are simultaneously carried out in each antibacterial test, and the result is shown in figure 1 ~ 6.

As shown in the comparison of the series of figures 1 ~ 6, the alginate composite antibacterial dressing prepared by the invention has excellent antibacterial property, and the antibacterial effect on Candida albicans can reach 100%.

Example 7

60 male New Zealand rabbits, 2 ~ 3kg each, were randomly divided into five groups of 12 rabbits each, each rabbit caused a 30% TBSA shallow II degree burn according to the standard model preparation method for clinical animal experiment burns, wherein group A rabbits did not use any dressing, group B rabbits used alginate dressing prepared by conventional processing without loading silk fibroin film, group C, D, E rabbits used alginate composite antibacterial dressing prepared by the invention of example 2 ~ 4, and changed twice a day until C, D, E groups of wounds healed, and the animal experiment was concluded.

TABLE 3 wound infection and healing (%)

As shown in Table 3, from the results of the animal experiment A ~ E group, the wounds of A, B two groups of rabbits were infected, while no infection occurred on the wounds of C, D, E three groups of rabbits after the alginate composite antibacterial dressing prepared in the invention example 2 ~ 4 was applied to the wounds of C, D, E three groups of rabbits, the healing time of the wounds of C, D, E three groups of rabbits was significantly shorter than that of the group B, and the time difference was significant.

From the overall results of five groups of experiments, the alginate dressing prepared by conventional processing without loading the silk fibroin film has a certain promotion effect on the healing of the wound surface, but compared with the alginate composite antibacterial dressing prepared by the invention, the alginate dressing prepared by conventional processing without loading the silk fibroin film has longer healing time for wound nursing, and the alginate composite antibacterial dressing prepared by the invention can obviously shorten the healing time of the wound and prevent the wound surface from being infected.

it should be noted that the specific embodiments are merely representative examples of the present invention, and it is obvious that the technical solution of the present invention is not limited to the above examples, and many variations are possible. Those skilled in the art, having the benefit of this disclosure and the benefit of this written description, will appreciate that other embodiments can be devised which do not depart from the specific details disclosed herein.

Claims

1. the alginate composite antibacterial dressing is characterized by comprising a silk fibroin composite membrane and alginate fibers;

The preparation method of the alginate composite antibacterial dressing comprises the following specific steps:

S1, preparation of alginate fibers: pre-dissolving alginate, stirring, filtering by adopting a filter cloth under the pressure of 0.1-0.3 Mpa to obtain alginate spinning solution, stretching, washing, dividing filaments and winding by adopting a wet spinning process and taking a calcium ion aqueous solution or an alcohol solution with the mass fraction of 5-10% as a coagulating bath, and drying under the hot air at the temperature of 100 ℃ to obtain alginate fibers;

S2, preparation of the silk fibroin solution: sequentially soaking natural silkworm shells or silks in a mixed organic solvent by adopting a salt-dissolving enzymolysis process, boiling the mixture in water for degumming, dissolving a mixed solution consisting of inorganic salt and formic acid, dialyzing, filtering and concentrating to obtain a silk fibroin solution with the mass concentration of 15-50%;

S3, preparation of the silk fibroin composite membrane: sequentially adding an organosilicon quaternary ammonium salt solution with the mass concentration of 12-20%, a sugar solution with the mass concentration of 1-10%, a hyaluronic acid solution with the mass concentration of 2-6%, a vitamin E solution with the mass concentration of 0.5-1%, and a hyaluronic acid solution with the mass concentration of 0.1-0.5% into the silk fibroin solution prepared in the step S2, fully stirring and mixing for 0.5-2 h, standing and defoaming to obtain a silk fibroin composite membrane mixed solution;

s4, loading of the silk fibroin composite membrane: adding 1-20% by mass of dihydric alcohol into the silk fibroin composite film mixed solution prepared in the step S3, wherein the volume ratio of the silk fibroin composite film mixed solution to ethylene glycol is 100: 1-10, padding the alginate fiber prepared in the step S1 in the mixed solution for 20-60 min under the conditions of constant temperature and constant humidity, taking out the alginate fiber, pre-freezing the alginate fiber at the low temperature of-80 to-20 ℃ for 10-24 h, and finally, drying the alginate fiber in vacuum for 18-24 h under the low temperature condition to obtain the membrane-carrying alginate composite fiber;

2. The alginate composite antibacterial dressing of claim 1, wherein the content of the silk fibroin composite membrane in the antibacterial dressing is 30-150 mg/g.

3. the alginate composite antibacterial dressing according to claim 1, wherein the pore size of the silk fibroin composite membrane is 20-150 μm.

4. The alginate composite antibacterial dressing of claim 1, wherein the thickness of the silk fibroin composite membrane in the antibacterial dressing is 1-30 μm.

5. The alginate composite antibacterial dressing of claim 1, wherein the pre-dissolving temperature in the step S1 is 30-40 ℃ and the time is 2-3 h; the stirring temperature is 30-60 ℃, the stirring speed is 200-400 r/min, and the stirring time is 3-7 h; the temperature of the wet spinning process is 60-80 ℃, the number of the spinneret holes is 200-500, and the spinning speed is 5-100 m/min.

6. The alginate composite antibacterial dressing of claim 1, wherein in the step S3, the ratio of the fibroin solution: organosilicon quaternary ammonium salt solution: sugar solution: hyaluronic acid solution: vitamin E solution: the volume ratio of the hyaluronic acid solution is 12-18: 2-6: 1-4: 4-8: 3-7: 2 to 5.

7. The alginate composite antimicrobial dressing of claim 6, wherein the silicone quaternary ammonium salt solution is one of quaternized haloalkylsilanes, quaternized amino silicone oils, quaternized hydrocarbon-based silicone oils; the sugar solution is one or more of glucose, lactose, galactose, mannose, ribose, sorbose, gulose, fucose, rhamnose, sucrose, fructose, maltose and cellobiose.

8. The alginate composite antibacterial dressing of claim 1, wherein the dihydric alcohol in the step S4 is one or more of ethylene glycol, 1, 3-propylene glycol, 2, 3-propylene glycol, 1, 4-butylene glycol, 1, 2-butylene glycol, 1, 3-butylene glycol, 2, 3-butylene glycol, diethylene glycol and triethylene glycol.