CN114702708A - Antibacterial material with wide pH application range and preparation method and application thereof - Google Patents
Antibacterial material with wide pH application range and preparation method and application thereof Download PDFInfo
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- CN114702708A CN114702708A CN202210359144.0A CN202210359144A CN114702708A CN 114702708 A CN114702708 A CN 114702708A CN 202210359144 A CN202210359144 A CN 202210359144A CN 114702708 A CN114702708 A CN 114702708A
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- sodium alginate
- polylysine
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- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 83
- 239000002131 composite material Substances 0.000 claims abstract description 75
- MGSRCZKZVOBKFT-UHFFFAOYSA-N thymol Chemical compound CC(C)C1=CC=C(C)C=C1O MGSRCZKZVOBKFT-UHFFFAOYSA-N 0.000 claims abstract description 48
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- OMNKZBIFPJNNIO-UHFFFAOYSA-N n-(2-methyl-4-oxopentan-2-yl)prop-2-enamide Chemical compound CC(=O)CC(C)(C)NC(=O)C=C OMNKZBIFPJNNIO-UHFFFAOYSA-N 0.000 claims abstract description 21
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
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Abstract
An antibacterial material with wide pH application range and a preparation method and application thereof are disclosed, which comprises the following steps: modifying sodium alginate by using a diacetone acrylamide solution to prepare a hydrophobic modified sodium alginate solution; respectively adding a polylysine solution and a modified sodium alginate solution into a 24-pore plate, fully reacting at the liquid surface junction to form a film-shaped product, cleaning, and freeze-drying to obtain the modified sodium alginate/polylysine composite film; catalyzing tannin solution to oxidize and self-polymerize by laccase to obtain tannin oligomer solution, then soaking the modified sodium alginate/polylysine composite film in the tannin oligomer solution, and cleaning to obtain the tannin-modified sodium alginate/polylysine composite material; soaking the tannin-modified sodium alginate/polylysine composite material in a thymol solution, and drying in a constant temperature and humidity box. The material has good antibacterial effect in a wide pH range, and the raw materials are mostly green natural substances, so that the material has good biocompatibility and simple preparation process.
Description
Technical Field
The invention relates to the technical field of antibacterial functional materials, in particular to an antibacterial material with a wide pH application range, and a preparation method and application thereof.
Background
With the development of society and the improvement of living standard of people, people pay more and more attention to the quality of life and sanitary conditions of people, and bacteria are common pathogenic microorganisms and influence the health of people, especially sick and wounded patients. Wound infection of injured patients is a major pathological factor causing the wound surface to be intractable, and in recent years, development and application of functional materials based on economic safety, durability and high-efficiency antibiosis are receiving more and more attention. The main antibacterial principle is to destroy the cell membrane structure of bacteria, cause the loss, damage or degeneration of substances in the cell membrane, influence the activity of cells and finally cause the death of microorganisms. The traditional antibacterial material mostly introduces metal or metal oxide nanoparticles or corresponding salts to achieve antibacterial property, but the environment for bacteria growth and propagation is complicated, and the change of pH value can occur due to different bacteria types. The problem that the pH action range of part of antibacterial materials on bacterial reproduction is narrow, namely, the antibacterial materials only have high-efficiency antibacterial property within a small pH range, and beyond the range, the antibacterial effect can be obviously weakened or even disappeared, and the problems of selectivity to drug-resistant strains, certain environmental toxicity, complex process and the like exist, so that the further application and development of the antibacterial materials are influenced, and therefore, the preparation of the antibacterial materials with good biocompatibility, broad-spectrum antibacterial property and wide pH application range has important significance.
Disclosure of Invention
The technical problem to be solved is as follows: aiming at the problems in the prior art, the invention provides an antibacterial material with a wide pH application range, a preparation method and application thereof, and the antibacterial material has the advantages of simple and convenient preparation process, environmental protection, no toxicity and the like, more importantly, the limitation of the pH action of the traditional antibacterial material is solved, and a new breakthrough is made on the antibacterial material with the wide pH application range.
The technical scheme is as follows: a preparation method of an antibacterial material with a wide pH application range comprises the following steps:
dissolving diacetone acrylamide (DAAM) in deionized water to prepare a diacetone acrylamide solution with the mass fraction of 10-15%, and fully stirring for 20 min;
dissolving Sodium Alginate (SA) in deionized water, preparing a sodium alginate solution with the mass fraction of 2-6%, and defoaming for 2-6 h by ultrasonic waves;
step three, sodium alginate is modified, the solution prepared in the step one and the solution prepared in the step two are mixed and placed in an oil bath pot for heating and stirring, and potassium persulfate (K) is added2S2O8) Adding potassium persulfate as initiator at 80-100 deg.C, and stirring for 2-4 hrThen cooling to room temperature and taking out to prepare modified sodium alginate (DSA);
dissolving Polylysine (PL) in deionized water to prepare a polylysine solution with the mass fraction of 4-8%;
step five, dropwise adding the modified sodium alginate and polylysine solution prepared in the step three and the step four into a 24-hole plate in sequence, allowing the liquid level to be layered obviously, standing for 8-12 h for reaction, taking out the film generated at the junction of the liquid level, cleaning for multiple times, and freeze-drying to obtain a modified sodium alginate/polylysine (DSA/PL) composite film;
step six, dissolving laccase in acetic acid-sodium acetate buffer solution with the pH value of 3-5, and stirring in ice bath for 30 min to prepare laccase solution with the concentration of 5-15 mg/mL;
dissolving tannic acid in a phosphate buffer solution with the pH =5.8, preparing a tannic acid solution with the mass fraction of 5-10%, and mixing the laccase solution prepared in the sixth step into the tannic acid solution for catalytic oxidation, wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 100-200 rpm, the time is 4-6 h, the mixture is taken out of the container and is shaken up manually to obtain the tannin oligomer solution (for better shaking up, the container can be taken out periodically for 1 h/time), and the pH of the solution is adjusted to be = 8.5;
step eight, placing the modified sodium alginate/polylysine composite film prepared in the step five into the tannic acid oligomer solution prepared in the step seven, soaking for 4-6 h, oscillating in a shaking table at the temperature of 30-40 ℃ at the speed of 100 plus 150 rpm, taking out, and cleaning to obtain the tannic acid-modified sodium alginate/polylysine (TA-DSA/PL) composite material;
and step nine, placing the tannin-modified sodium alginate/polylysine composite material prepared in the step eight in a 1-5 wt% Thymol solution for soaking for 4-6 h (glycerol can be properly added for promoting dissolution), shaking the mixture in a shaking table at the temperature of 30-40 ℃ at the speed of 100-150 rpm, taking out the mixture, cleaning the mixture, and drying the mixture in a constant temperature and humidity chamber to obtain the Thymol-tannin-modified sodium alginate/polylysine (Thymol-TA-DSA/PL) composite material with the antibacterial property in a wide pH range.
Preferably, the ratio of the diacetone acrylamide solution to the sodium alginate solution to the potassium persulfate in the step three is 20 mL: 20-60 mL: 5-15 mg.
Preferably, the volume ratio of the modified sodium alginate solution to the polylysine solution in the fifth step is 1: 1.
preferably, the volume ratio of the laccase solution to the tannic acid solution in the seventh step is 1 (3-5).
Preferably, the temperature range of the constant temperature and humidity box in the ninth step is 20-37 ℃, and the humidity is 30-50% RH.
The antibacterial material prepared by the method has wide pH application range.
The antibacterial material with wide pH application range is applied to preparing wound antibacterial materials.
Has the advantages that: (1) the invention takes the modified sodium alginate/polylysine composite film with good biocompatibility as a base material, and the sodium alginate is subjected to hydrophobic modification in advance to prepare the amphiphilic sodium alginate, so that the purpose of stable size in a solution environment is achieved, and the loading rate and the fixed amount of the antibacterial agent are improved;
(2) according to the invention, the polyphenol oxidase laccase is adopted to catalyze the tannic acid, so that the tannic acid is self-polymerized to form tannic acid oligomer, and the antibacterial effect of the tannic acid is improved. The tannin oligomer is fixed on the modified sodium alginate/polylysine film through Schiff base action or Michael addition reaction, and the load is firm.
(3) The selected antibacterial agent is two natural antibacterial agents of tannic acid and thymol with good antibacterial effect in different pH environments, has the advantages of broad-spectrum antibacterial property, high efficiency and durability, good biocompatibility, wide sources and the like, has the antibacterial rate of more than 98 percent and the killing effect of 99.99 percent at most on gram-negative bacteria and gram-positive bacteria within the range of pH =3-8, and has excellent antibacterial effect.
(4) The preparation method disclosed by the invention is simple in preparation process, convenient to operate and environment-friendly.
Drawings
FIG. 1 is a surface microstructure and a cross-sectional view of an antibacterial material substrate according to the present invention;
FIG. 2 is a graph of the swelling weight gain of the antibacterial material of the present invention in different solution environments;
FIG. 3 is a graph showing the antibacterial effect of the antibacterial material according to the present invention;
FIG. 4 is a schematic representation of the cytotoxicity of the invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings and specific embodiments.
Example 1
The antibacterial material with the wide pH application range and the antibacterial characteristic comprises a modified sodium alginate/polylysine composite film base material, wherein the modified sodium alginate/polylysine composite film is treated to obtain the antibacterial material loaded with tannic acid and thymol, and the antibacterial material has the wide pH application range.
The preparation method of the antibacterial material with the wide pH range antibacterial characteristic comprises the steps of sodium alginate modification, film substrate preparation and antibacterial agent loading, and specifically comprises the following steps:
s1, dissolving diacetone acrylamide in deionized water to prepare a diacetone acrylamide solution with the mass fraction of 10%, and fully stirring for 20 min;
s2, dissolving sodium alginate in deionized water, preparing a sodium alginate solution with the mass fraction of 6%, and defoaming for 6 hours by ultrasonic waves;
s3, mixing the solutions prepared from S1 and S2 according to the volume ratio of 1:1, heating and stirring the mixture in an oil bath pot (20 mL of diacetone acrylamide solution is taken in the embodiment), adding 5mg of potassium persulfate, stirring the mixture for 3 hours at the temperature of 90 ℃, cooling the mixture to room temperature, and taking the mixture out to prepare modified sodium alginate DSA;
s4, dissolving polylysine in deionized water to prepare a polylysine solution with the mass fraction of 8%;
s5, sequentially dripping the solutions prepared from S3 and S4 into a 24-hole plate according to the volume ratio of 1:1, obviously layering liquid surfaces, standing for reaction for 10 hours, taking out a film generated at the junction of the liquid surfaces, cleaning for multiple times, and freeze-drying to obtain a DSA/PL composite film;
s6, dissolving laccase in an acetic acid-sodium acetate buffer solution with the pH =3, and stirring for 30 min in an ice bath to prepare a laccase solution with the concentration of 5 mg/mL;
s7, dissolving tannic acid in phosphate buffer solution with pH =5.8, preparing tannic acid solution with mass fraction of 10%, adding the laccase solution prepared in S6 into the tannic acid solution for catalytic oxidation (V laccase solution: V tannic acid solution =1: 3), wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 200 rpm, the time is 6 h, the mixture is taken out of the container periodically (1 h/time) and is shaken up manually to obtain a tannin oligomer solution, and the pH of the solution is adjusted to be = 8.5;
s8, placing the modified sodium alginate/polylysine composite film prepared in the step S5 in the tannin oligomer solution prepared in the step S7, soaking for 6 hours, oscillating in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out and cleaning to obtain a TA-DSA/PL composite material;
s9, placing the tannin-modified sodium alginate/polylysine composite material prepared in the step S8 into a 1 wt% Thymol solution for soaking for 6 h (glycerol can be properly added to promote dissolution), shaking the mixture in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out the mixture, washing the mixture, and drying the mixture in a constant temperature and humidity box (37 ℃ and 50% RH) to obtain Thymol-TA-DSA/PL with the antibacterial property in a wide pH range.
Example 2
The antibacterial material with the wide pH application range and the antibacterial characteristic comprises a modified sodium alginate/polylysine composite film base material, wherein the modified sodium alginate/polylysine composite film is treated to obtain the antibacterial material loaded with tannic acid and thymol, and the antibacterial material has the wide pH application range.
The preparation method of the antibacterial material with the wide pH range antibacterial characteristic comprises the steps of sodium alginate modification, film substrate preparation and antibacterial agent loading, and specifically comprises the following steps:
s1, dissolving diacetone acrylamide in deionized water to prepare a diacetone acrylamide solution with the mass fraction of 15%, and fully stirring for 20 min;
s2, dissolving sodium alginate in deionized water, preparing a sodium alginate solution with the mass fraction of 2%, and defoaming for 2 hours by ultrasonic waves;
s3, mixing the solutions prepared from S1 and S2 according to the volume ratio of 1:3, heating and stirring the mixture in an oil bath pot (20 mL of diacetone acrylamide solution is taken in the embodiment), adding 15mg of potassium persulfate, stirring the mixture for 2 hours at the temperature of 80 ℃, cooling the mixture to room temperature, and taking the mixture out to prepare modified sodium alginate DSA;
s4, dissolving polylysine in deionized water to prepare a polylysine solution with the mass fraction of 4%;
s5, sequentially dripping the solutions prepared from S3 and S4 into a 24-hole plate according to the volume ratio of 1:1, obviously layering liquid surfaces, standing for reaction for 8 hours, taking out a film generated at the junction of the liquid surfaces, cleaning for multiple times, and freeze-drying to obtain a DSA/PL composite film;
s6, dissolving laccase in an acetic acid-sodium acetate buffer solution with the pH =5, and stirring for 30 min in an ice bath to prepare a laccase solution with the concentration of 15 mg/mL;
s7, dissolving tannic acid in phosphate buffer solution with pH =5.8, preparing tannic acid solution with mass fraction of 5%, adding the laccase solution prepared in S6 into the tannic acid solution for catalytic oxidation (V laccase solution: V tannic acid solution =1: 5), wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 150 rpm, the time is 4h, the mixture is taken out of the container periodically (1 h/time) and is shaken up manually to obtain a tannin oligomer solution, and the pH of the solution is adjusted to be = 8.5;
s8, placing the modified sodium alginate/polylysine composite film prepared in the step S5 in the tannin oligomer solution prepared in the step S7, soaking for 4 hours, oscillating in a shaking table at the temperature of 40 ℃ at the speed of 150 rpm, taking out, and cleaning to obtain a TA-DSA/PL composite material;
s9, placing the tannin-modified sodium alginate/polylysine composite material prepared in the step S8 into a 2.5 wt% Thymol solution for soaking for 4 hours (glycerol can be properly added for promoting dissolution), shaking the composite material in a shaking table at the temperature of 40 ℃ at the speed of 150 rpm, taking out the composite material, cleaning the composite material, and drying the composite material in a constant temperature and humidity box to obtain Thymol-TA-DSA/PL with the antibacterial property of a wide pH range.
Example 3
The antibacterial material with the wide pH application range and the antibacterial characteristic comprises a modified sodium alginate/polylysine composite film base material, wherein the modified sodium alginate/polylysine composite film is treated to obtain the antibacterial material loaded with tannic acid and thymol, and the antibacterial material has the wide pH application range.
The preparation method of the antibacterial material with the wide pH range antibacterial characteristic comprises the steps of sodium alginate modification, film substrate preparation and antibacterial agent loading, and specifically comprises the following steps:
s1, dissolving diacetone acrylamide in deionized water to prepare a diacetone acrylamide solution with the mass fraction of 12.5%, and fully stirring for 20 min;
s2, dissolving sodium alginate in deionized water, preparing a sodium alginate solution with the mass fraction of 4%, and defoaming for 4 hours by ultrasonic waves;
s3, mixing the solutions prepared from S1 and S2 according to the volume ratio of 1:2, heating and stirring the mixture in an oil bath pot (20 mL of diacetone acrylamide solution is taken in the embodiment), adding 10 mg of potassium persulfate, stirring the mixture for 4 hours at the temperature of 100 ℃, cooling the mixture to room temperature, and taking the mixture out to prepare modified sodium alginate DSA;
s4, dissolving polylysine in deionized water to prepare a polylysine solution with the mass fraction of 6%;
s5, sequentially dripping the solutions prepared from S3 and S4 into a 24-hole plate according to the volume ratio of 1:1, obviously layering liquid surfaces, standing for reaction for 12 hours, taking out a film generated at the junction of the liquid surfaces, cleaning for multiple times, and freeze-drying to obtain a DSA/PL composite film;
s6, dissolving laccase in an acetic acid-sodium acetate buffer solution with the pH =4, and stirring for 30 min in an ice bath to prepare a laccase solution with the concentration of 5 mg/mL;
s7, dissolving tannic acid in phosphate buffer solution with pH =5.8, preparing tannic acid solution with mass fraction of 7.5%, adding the laccase solution prepared by S6 into the tannic acid solution for catalytic oxidation (V laccase solution: V tannic acid solution =1: 4), wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 100 rpm, the time is 5 h, the mixture is taken out of the container periodically (1 h/time) and is shaken up manually to obtain a tannin oligomer solution, and the pH of the solution is adjusted to be = 8.5;
s8, placing the modified sodium alginate/polylysine composite film prepared in the step S5 in the tannin oligomer solution prepared in the step S7, soaking for 5 hours, oscillating in a shaking table at the temperature of 35 ℃ at the speed of 125 rpm, taking out, and cleaning to obtain a TA-DSA/PL composite material;
s9, placing the tannin-modified sodium alginate/polylysine composite material prepared in the step S8 into a 5 wt% Thymol solution for soaking for 5 hours (glycerol can be properly added for dissolution promotion), shaking the mixture in a shaking table at the temperature of 35 ℃ at the speed of 125 rpm, taking out the mixture, washing the mixture, and drying the mixture in a constant temperature and humidity box to obtain Thymol-TA-DSA/PL with the antibacterial property of wide pH range.
Comparative example 1
The comparative example specifically included the following steps:
s1, dissolving diacetone acrylamide in deionized water, preparing 20mL of a diacetone acrylamide solution with the mass fraction of 10%, and fully stirring for 20 min;
s2, dissolving sodium alginate in deionized water, preparing a sodium alginate solution with the mass fraction of 6%, and defoaming for 6 hours by ultrasonic waves;
s3, mixing the solutions prepared from S1 and S2 according to the volume ratio of 1:1, heating and stirring the mixture in an oil bath pot, adding 5mg of potassium persulfate at the temperature of 90 ℃, stirring the mixture for 3 hours, cooling the mixture to room temperature, and taking the mixture out to obtain modified sodium alginate DSA;
s4, dissolving polylysine in deionized water to prepare a polylysine solution with the mass fraction of 8%;
s5, sequentially dripping the solutions prepared from S3 and S4 into a 24-hole plate according to the volume ratio of 1:1, obviously layering liquid surfaces, standing for reaction for 10 hours, taking out a film generated at the junction of the liquid surfaces, cleaning for multiple times, and freeze-drying to obtain a DSA/PL composite film;
s6, dissolving laccase in an acetic acid-sodium acetate buffer solution with the pH =3, and stirring for 30 min in an ice bath to prepare a laccase solution with the concentration of 5 mg/mL;
s7, dissolving tannic acid in phosphate buffer solution with pH =5.8, preparing tannic acid solution with mass fraction of 10%, adding the laccase solution prepared in S6 into the tannic acid solution for catalytic oxidation (V laccase solution: V tannic acid solution =1: 3), wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 200 rpm, the time is 6 h, the mixture is taken out of the container periodically (1 h/time) and is shaken up manually to obtain a tannin oligomer solution, and the pH of the solution is adjusted to be = 8.5; dissolving a thymol solution in deionized water to prepare a thymol solution with the mass fraction of 1%;
s8, placing the modified sodium alginate/polylysine composite film prepared in the step S5 in the tannin oligomer solution prepared in the step S7, soaking for 6 hours, oscillating in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out, and cleaning to obtain a TA-DSA/PL composite material; in addition, placing the modified sodium alginate/polylysine composite film prepared by S5 with the same amount in Thymol solution prepared by S7, soaking for 6 h, shaking by a shaker at the temperature of 30 ℃ at the speed of 100 rpm, taking out and washing to obtain Thymol-DSA/PL composite material;
s9, placing the tannin-modified sodium alginate/polylysine (TA-DSA/PL) composite material prepared in the S8 into the Thymol solution prepared in the S7, soaking for 6 hours, shaking the composite material in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out the composite material, cleaning, and drying the composite material in a constant temperature and humidity box (37 ℃ and 50% RH) to obtain Thymol-TA-DSA/PL with the antibacterial property of wide pH range.
Comparative example 2
According to the comparative example, unmodified sodium alginate and polylysine are used for preparing the composite film base material, and the sodium alginate/polylysine composite film is treated in the same way as in the examples to obtain the tannin and thymol loaded antibacterial material. The method specifically comprises the following steps:
s1, dissolving sodium alginate in deionized water, preparing a sodium alginate solution with the mass fraction of 6%, and defoaming for 6 hours by ultrasonic waves;
s2, dissolving polylysine in deionized water to prepare a polylysine solution with the mass fraction of 8%;
s3, sequentially dropwise adding the solutions prepared from S1 and S2 into a 24-hole plate according to the volume ratio of 1:1, obviously layering liquid surfaces, standing for reaction for 10 hours, taking out a film generated at the junction of the liquid surfaces, washing for multiple times, and freeze-drying to obtain an SA/PL composite film;
s4, dissolving laccase in an acetic acid-sodium acetate buffer solution with the pH =3, and stirring for 30 min in an ice bath to prepare a laccase solution with the concentration of 5 mg/mL;
s5, dissolving tannic acid in phosphate buffer solution with pH =5.8, preparing tannic acid solution with mass fraction of 10%, adding the laccase solution prepared in S6 into the tannic acid solution for catalytic oxidation (V laccase solution: V tannic acid solution =1: 3), wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 200 rpm, the time is 6 h, the container is taken out every hour, the mixture is shaken up by hand to obtain the tannin oligomer solution, and the pH of the solution is adjusted to be = 8.5; dissolving a thymol solution in deionized water to prepare a thymol solution with the mass fraction of 1%;
s6, placing the sodium alginate/polylysine composite film prepared in the step S5 in the tannin oligomer solution prepared in the step S7, soaking for 6 hours, oscillating in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out and washing to obtain the TA-SA/PL composite material;
s7, placing the sodium alginate/polylysine composite material prepared in the step S3 in the Thymol solution prepared in the step S5, soaking for 6 hours, shaking the mixture in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out the mixture, and washing the mixture to obtain a Thymol-SA/PL composite material; and (2) placing the tannin-sodium alginate/polylysine composite material prepared in the step (S6) into the Thymol solution prepared in the step (S5) for soaking for 6 h, shaking the composite material in a shaking table at the temperature of 30 ℃ at the speed of 100 rpm, taking out and washing the composite material to obtain the Thymol-TA-SA/PL composite material, and drying the composite material in a constant temperature and humidity chamber (37 ℃ and 50% RH) to obtain Thymol-TA-SA/PL with the antibacterial property in a wide pH range.
The DSA/PL composite film prepared in example 1, the TA-DSA/PL composite material prepared in example 1, the Thymol-DSA/PL composite material prepared in comparative example 1, the SA/PL composite film prepared in comparative example 2, the Thymol-SA/PL composite material prepared in comparative example 2, the TA-SA/PL composite material prepared in comparative example 2, and the Thymol-TA-SA/PL composite material prepared in comparative example 2 were tested by the following specific test procedures:
microscopic morphology: surface topography images of the composite films were taken by Scanning Electron Microscopy (SEM) using Hitachi SU1510 at 5.0 kV.
FIG. 1 is a surface microstructure and a cross-sectional view of a DSA/PL thin film substrate prepared in example 1 of the present invention, wherein (a) is a DSA/PL surface topography, (b) is an enlarged view of the graph surface, and (c) is a DSA/PL cross-sectional view, from which it can be seen that the surface of the DSA/PL film prepared by DAAM hydrophobic modification of SA is porous and has a certain network structure, because the viscosity of the modified DSA is reduced and the contact with PL solution is more sufficient. From the cross-section, it can be observed that the film exhibits a multilayer irregular stack structure with a thickness of about 210 + -4 μm.
Swelling property test: the test was performed in deionized water, phosphate buffered saline (PBS, pH =7.4 ± 0.2), and HCl (pH =1.5) media, respectively, at room temperature, to study the extent to which the composite membrane absorbs moisture and swells over time. Before testing, the samples were cut to the same size and placed in a 10% RH incubator for 24 hours, then weighed immediately (M 0) And then immersed in deionized water. The weight of the samples was recorded at various time intervals (t), i.e. 0.5, 1, 2, 3, 4, 5, 6, 7 hours, first blotting the surface of the film with soft paper to remove excess moisture and then weighing immediately. The testing procedure was the same for PBS and HCl solution. The swelling ratio (weight gain) was calculated according to the following formula (1), whereinM t = sample weight at soaking time t,M 0 = initial weight.
Swelling ratio% = (b) (%) (ii)M t-M 0)/M 0×100 （1）
Fig. 2 is a graph showing the swelling weight gain of the antibacterial material of the present invention, and it can be seen that the swelling weight gain of the antibacterial material of the present invention after hydrophobic modification with diacetone acrylamide in PBS buffer, water, and HCl solution with pH =1.5 is significantly reduced, and the dimensional stability is greatly improved.
And (3) testing the antibacterial effect: and (3) performing antibacterial performance evaluation on the material by referring to AATCC100-2012 'evaluation method of antibacterial textiles', and selecting staphylococcus aureus in gram-positive bacteria and escherichia coli in gram-negative bacteria as test strains. The material was cut into several round samples of consistent size and thickness and placed in a 24-well plate. Taking 0.1 mL of 108 - 109CFU/mL of the bacterial solution was dropped on the samples in the 24-well plate, and after contacting the samples for 40 min, 0.9 mL of Phosphate Buffered Saline (PBS) corresponding to the pH was added. After shaking uniformly, 0.1 mL of original bacteria liquid and bacteria liquid contacted with the sample are respectively diluted in a centrifugal tube in a medium gradient way by 10６Fold (3 parallel experiments were done for each sample). Dripping 10 μ L of each dilution gradient solution on a culture medium plate, culturing in a 37 deg.C incubator for 24 hr, and recordingThe colony number of each row of the plate is recorded, and the antibacterial effect of the composite membrane is evaluated by calculating the bacterial survival rate (N,%) according to the formula (2).
N%＝Ni/N0×100 （2）
In the formula, N0Counting the maximum value of the bacterial colony in the original bacterial liquid; n is a radical ofiThe bacterial colony value of the sample after antibiosis and corresponding gradient of the original bacterial liquid is shown.
The antibacterial effect test is shown in fig. 3 (wherein s. aureus-staphylococcus aureus, gram positive bacteria; e. coli, gram negative bacteria; DSA/PL-modified sodium alginate/polylysine composite film; TA-DSA/PL-composite film loaded with tannic acid alone; Thymol-TA-DSA/PL-composite film loaded with Thymol alone; Thymol-TA-DSA/PL-antibacterial material with wide pH range), four samples are put into a 24-well plate in triplicate, and are placed for 1 hour after bacterial liquid is added, and then an antibacterial test is performed, wherein (a) when two pH environments (pH =4.5, pH = 7.5) are set for the test, the antibacterial effectiveness of the antibacterial material in an acid-base environment is detected: test results show that DSA/PL has certain bactericidal effect on staphylococcus aureus and escherichia coli under the conditions of pH =4.5 and pH =7.5, but cannot be applied to high-efficiency antibacterial materials; tannin and DSA/PL can generate Schiff base combination or Michael addition under an alkaline environment, so that the antibacterial effect is better when the pH is = 7.5; the thymol is combined with DSA/PL through hydrogen bonds, so that the antibacterial effect is better in an acid environment; meanwhile, DSA/PL loaded with tannic acid and thymol has a good killing effect on staphylococcus aureus and escherichia coli, and the sterilization rates are 99.28% and 99.48% respectively when the pH is = 4.5; at pH =7.5, the bactericidal rate was 99.57% and 99.98%, respectively. (b) The antibacterial effect of Thymol-TA-DSA/PL under a wide pH range is explored for expanding the pH range, and the highest antibacterial effect can reach 99.99%.
The antibacterial effect is compared with that shown in the following table, and it can be seen from the table that the modified DSA/PL antibacterial effect is greatly improved, and the combination with thymol is enhanced after hydrophobic modification, so that the antibacterial effect of thymol is shown.
And (3) testing the biocompatibility:
cell culture
L929 mouse skin fibroblasts were selected for in vivo cytotoxicity studies and cultured in a complete cell culture medium (MEM) prepared at a ratio of 9:1 horse serum. Cells were incubated at 5% CO2Incubated at 37 ℃ in a humidified atmosphere and the medium was changed every 1-2 days. Third generation L929 cells were used for all experiments.
Cell viability study
L929 cells were seeded into 96-well plates and then incubated overnight. The area is about 1.5 cm2Is selected from the group consisting of SA/PL, Thymol-SA/PL, TA-SA/PL, Thymol-TA-SA/PL, DSA/PL, Thymol-DSA/PL, TA-DSA/PL and Thymol-TA-DSA/PL, and the film is sterilized by ultraviolet irradiation for 2 hours. MEM was added to the sterilized samples for 24 hours of leaching according to ISO 10993-5:1999, then 100 μ L of the medium extract was transferred to each well, the cells were further cultured for 24 hours, and their viability was determined by MTT method: taking L929 cells in logarithmic growth phase, counting cells, adjusting cell concentration according to 8 × 103Perwell inoculation into 96-well plates, 5% CO2And culturing overnight in a constant-temperature incubator at 37 ℃ until the cells adhere to the wall. The 8 samples were processed in groups and incubated for 24 h. The medium containing the sample is removed. Each well was washed three times with PBS and 100. mu.L of medium containing 0.5 mg/mL MTT, 5% CO per well2And culturing in a constant temperature incubator at 37 ℃ for 4 hours. The supernatant was discarded and 100. mu.L DMSO was added to each well. After shaking gently for 10 min, the absorbance at 570 nm was measured. Cell viability was calculated according to equation (3):
cell viability% = amembrane⁄Acontrol×100 （3）
In addition, LIVE/DEAD stain was used in order to visually observe the viability of adherent cells on the sample. After 24h incubation, cells on the sample were washed 3 times with PBS. Then 100. mu.L of the staining solution was dropped onto the sample and after incubation for 1h at 37 ℃ in the dark, studies were performed on green viable cells and red dead cells using CLSM.
FIG. 4 is a schematic diagram showing cytotoxicity of the antibacterial material, in which (a) shows L929 cell viability measurement by the MTT method, and (b) shows fluorescence imaging (LIVE/DEAD staining) of L929 cells. Thymol-TA-SA/PL and Thymol-TA-DSA/PL membranes showed normal cell proliferation (no toxicity to L929 cells), SA/PL, Thymol-SA/PL, DSA/PL and Thymol-DSA/PL promoted cell proliferation, while TA-SA/PL and TA-DSA/PL were slightly cytotoxic. Furthermore, fluorescence imaging (LIVE/DEAD staining) of L929 cells cultured on the extract for 24 hours as shown in figure (b), all samples except TA-DSA/PL consistent with the MTT cell viability assay can support cell growth without significant cytotoxicity. These results indicate that Thymol-TA-DSA/PL membrane has excellent biocompatibility.
Claims (7)
1. A preparation method of an antibacterial material with a wide pH application range is characterized by comprising the following steps:
dissolving diacetone acrylamide in deionized water, preparing a diacetone acrylamide solution with the mass fraction of 10-15%, and fully stirring for 20 min;
dissolving sodium alginate in deionized water, preparing a sodium alginate solution with the mass fraction of 2-6%, and defoaming for 2-6 h by ultrasonic;
step three, modifying sodium alginate, mixing the solution prepared in the step one and the solution prepared in the step two, heating and stirring the mixture in an oil bath pot, adding potassium persulfate serving as an initiator into the mixture at the temperature of between 80 and 100 ℃, stirring the mixture for 2 to 4 hours, cooling the mixture to room temperature, and taking the mixture out to prepare modified sodium alginate;
dissolving polylysine in deionized water to prepare a polylysine solution with the mass fraction of 4-8%;
step five, dropwise adding the modified sodium alginate and polylysine solution prepared in the step three and the step four into a 24-hole plate in sequence, obviously layering the liquid surface, standing for 8-12 h for reaction, taking out the film generated at the junction of the liquid surface, washing for multiple times, and freeze-drying to obtain the modified sodium alginate/polylysine composite film;
step six, dissolving laccase in acetic acid-sodium acetate buffer solution with the pH value of 3-5, and stirring in ice bath for 30 min to prepare laccase solution with the concentration of 5-15 mg/mL;
dissolving tannic acid in a phosphate buffer solution with the pH =5.8, preparing a tannic acid solution with the mass fraction of 5-10%, and mixing the laccase solution prepared in the sixth step into the tannic acid solution for catalytic oxidation, wherein the catalytic conditions are as follows: under the constant temperature condition of 30 ℃, the shaking table speed is 100-200 rpm, the time is 4-6 h, the container is taken out and shaken evenly by hand to obtain the tannin oligomer solution, and the pH of the solution is adjusted to be = 8.5;
step eight, placing the modified sodium alginate/polylysine composite film prepared in the step five into the tannic acid oligomer solution prepared in the step seven, soaking for 4-6 h, oscillating in a shaking table at the temperature of 30-40 ℃ at the speed of 100-150 rpm, taking out, and cleaning to obtain the tannic acid-modified sodium alginate/polylysine composite material;
and step nine, placing the tannin-modified sodium alginate/polylysine composite material prepared in the step eight in a 1-5 wt% thymol solution for soaking for 4-6 h, shaking the composite material in a shaking table at the temperature of 30-40 ℃ at the speed of 100-150 rpm, taking out the composite material, cleaning the composite material, and drying the composite material in a constant temperature and humidity box to obtain the thymol-tannin-modified sodium alginate/polylysine composite material with the antibacterial property in a wide pH range.
2. The method for preparing the antibacterial material with a wide pH application range according to claim 1, wherein the ratio of diacetone acrylamide solution, sodium alginate solution and potassium persulfate in the third step is 20 mL: 20-60 mL: 5-15 mg.
3. The method for preparing the antibacterial material with wide pH application range according to claim 1, wherein the volume ratio of the modified sodium alginate solution to the polylysine solution in the fifth step is 1: 1.
4. the method for preparing the antibacterial material with the wide pH application range according to claim 1, wherein the volume ratio of the laccase solution to the tannin solution in the seventh step is 1 (3-5).
5. The preparation method of the antibacterial material with the wide pH application range according to claim 1, wherein in the ninth step, the temperature range of the constant temperature and humidity chamber is set to be 20-37 ℃, and the humidity is 30-50% RH.
6. An antibacterial material having a wide pH application range prepared by the method of any one of claims 1 to 5.
7. The use of the antibacterial material with a wide pH application range of claim 6 in the preparation of antibacterial materials for wound surfaces.
Priority Applications (1)
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115887793A (en) * | 2022-10-08 | 2023-04-04 | 东华大学 | Preparation and amination method of polyphenol oxidase catalyzed polyphenol coating material |
| CN116350833A (en) * | 2023-04-14 | 2023-06-30 | 中国科学院长春应用化学研究所 | Polylysine-based antibacterial medical bone tissue adhesive and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977534A (en) * | 2011-09-07 | 2013-03-20 | 江南大学 | Preparation method for sodium alginate and poly (diacetone acrylamide) interpenetrating network gel |
| CN104939269A (en) * | 2015-06-16 | 2015-09-30 | 宁波大学 | Edible composite sodium alginate antibacterial film liquid as well as preparation method and application thereof |
| CN107484814A (en) * | 2017-07-06 | 2017-12-19 | 浦江县欧立生物技术有限公司 | Composite film-forming preservative |
| CN111996616A (en) * | 2020-08-20 | 2020-11-27 | 江南大学 | Sodium alginate/polylysine self-assembled fiber with controllable size |
-
2022
- 2022-04-06 CN CN202210359144.0A patent/CN114702708B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977534A (en) * | 2011-09-07 | 2013-03-20 | 江南大学 | Preparation method for sodium alginate and poly (diacetone acrylamide) interpenetrating network gel |
| CN104939269A (en) * | 2015-06-16 | 2015-09-30 | 宁波大学 | Edible composite sodium alginate antibacterial film liquid as well as preparation method and application thereof |
| CN107484814A (en) * | 2017-07-06 | 2017-12-19 | 浦江县欧立生物技术有限公司 | Composite film-forming preservative |
| CN111996616A (en) * | 2020-08-20 | 2020-11-27 | 江南大学 | Sodium alginate/polylysine self-assembled fiber with controllable size |
Non-Patent Citations (2)
| Title |
|---|
| XIN HUANG ET AL.: ""Electrostatically self-assembled filamentous sodium alginate/ε-polylysine fiber with antibacterial, bioadhesion and biocompatible in suturing wound"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| 蓝虹云 等: ""漆树漆酶催化单宁酸的氧化反应研究"", 《化学与生物工程》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115887793A (en) * | 2022-10-08 | 2023-04-04 | 东华大学 | Preparation and amination method of polyphenol oxidase catalyzed polyphenol coating material |
| CN116350833A (en) * | 2023-04-14 | 2023-06-30 | 中国科学院长春应用化学研究所 | Polylysine-based antibacterial medical bone tissue adhesive and preparation method and application thereof |
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