CN113440647A

CN113440647A - Preparation method and application of ozone hydrogel dressing

Info

Publication number: CN113440647A
Application number: CN202110864445.4A
Authority: CN
Inventors: 鲁建云; 袁小川; 周强; 喻小春
Original assignee: Hunan Haizhi Medical Technology Co ltd; Third Xiangya Hospital of Central South University
Current assignee: Hunan Haizhi Medical Technology Co ltd; Third Xiangya Hospital of Central South University
Priority date: 2021-07-29
Filing date: 2021-07-29
Publication date: 2021-09-28

Abstract

The invention discloses a preparation method and application of an ozone hydrogel dressing, belonging to the technical field of preparation of medical dressings, wherein the preparation method of the dressing comprises the following steps: firstly, weighing raw materials; step two, mixing the modified agar and 50% of deionized water, heating and stirring until the modified agar is completely dissolved, then stopping heating, stirring and cooling to obtain a first mixture; and thirdly, mixing the additive and the polyethylene glycol uniformly, adding the rest deionized water to obtain a second mixture, adding the second mixture into the first mixture, introducing ozone while stirring, then pouring into a culture dish with grid cloth, and performing gamma-ray irradiation to obtain the ozone water gel dressing. The hydrogel has excellent performance and good biological safety, and is a novel wound application and liquid medicine application auxiliary material with excellent performance. The prepared ozone water gel dressing can be applied to the moist healing of wounds.

Description

Preparation method and application of ozone hydrogel dressing

Technical Field

The invention belongs to the technical field of medical dressing preparation, and particularly relates to a preparation method and application of an ozone hydrogel dressing.

Background

The hydrogel is a hydrophilic reticular polymer swelling body containing a large amount of water, the dressing has good biocompatibility and hydrophilicity, can continuously release water outwards under certain conditions, can provide a wet healing environment for wounds, has a smooth surface, does not adhere to the wounds, cannot cause secondary damage to the wounds, and also has certain cooling and pain relieving effects.

Most hydrogel dressings do not have an antibacterial effect, so that medicines such as different antibacterial agents, bioactive components and the like need to be embedded in hydrogel according to different needs, and the medicines can be slowly and continuously released to a lesion area through body fluid exchange. However, the drug is difficult to control in combination with the hydrogel matrix and release thereof by means of embedding, so that the expected bacteriostatic effect cannot be achieved, and the embedding method is complex in manufacturing process.

Disclosure of Invention

The invention provides a preparation method and application of an ozone hydrogel dressing.

The purpose of the invention can be realized by the following technical scheme:

a preparation method of the ozone hydrogel dressing comprises the following steps:

firstly, weighing the following raw materials in parts by weight: 0.5-2 parts of modified agar, 1-20 parts of polyethylene glycol, 2-20 parts of additive and 60-85 parts of deionized water;

secondly, mixing the modified agar with 50 percent of deionized water, heating and stirring the mixture at the temperature of 95-100 ℃ and the rotating speed of 400-500r/min until the modified agar is completely dissolved, then stopping heating, stirring and cooling the mixture to the temperature of 40 ℃ to obtain a first mixture;

step three, mixing the additive and polyethylene glycol uniformly, then adding the rest deionized water, stirring and mixing at the rotation speed of 400-500r/min to obtain a second mixture, adding the second mixture into the first mixture, introducing ozone while stirring, controlling the system temperature at 25-35 ℃, the pressure at 0.5-1.5bar and the ozone content at 80-120g/L, then pouring into a culture dish with grid cloth, and carrying out gamma ray irradiation, wherein the irradiation dose is as follows: 30-90kGy, and the irradiation time is 0.5-2h to obtain the ozone hydrogel dressing.

The modified agar is prepared by the following steps:

mixing agar powder and deionized water, stirring at the temperature of 35 ℃, adjusting the pH value to 9 by using 1mol/L sodium hydroxide solution, then adding a mixed solution of a modifier and isopropanol, finishing dropwise adding the mixed solution within 1.5h, keeping the temperature unchanged after dropwise adding, continuing stirring for 3h, adjusting the pH value to 6 by using 1mol/L hydrochloric acid solution, performing post-treatment after the reaction is finished, wherein the post-treatment process comprises the following steps: and carrying out vacuum filtration on the obtained reaction liquid, washing a filter cake twice by using an ethanol water solution with the volume fraction of 60% and deionized water in sequence, drying the filter cake to constant weight at 50 ℃ after washing, and then crushing and sieving the filter cake by using a 100-mesh sieve to obtain the modified agar.

Further, the using amount ratio of the agar powder to the deionized water is 3-10 g: 100mL, and the dosage ratio of the mixed solution of the modifier and the isopropanol is 1 g: 10 mL; the dosage ratio of the agar powder to the mixed liquid of the modifier and the isopropanol is 1-3 g: 100 mL.

Further, the modifier is prepared by the following steps:

step S11, mixing 2-octenyl succinic anhydride and methanol, stirring to be transparent at the temperature of 45 ℃, then heating to 60 ℃, reacting for 2 hours, then adding anhydrous aluminum trichloride, reacting for 2.5 hours at the temperature of 80 ℃, and performing post-treatment after the reaction is finished, wherein the post-treatment process comprises the following steps: recrystallizing the obtained reaction solution with deionized water at 75 ℃, then cooling and filtering, and drying at 55 ℃ to constant weight to obtain an intermediate 1;

the reaction process is as follows:

step S12, dripping L-ascorbic acid and the intermediate 1 into a sulfuric acid solution, stirring for 1h at the temperature of 20 ℃, reacting for 28h at the temperature of 25 ℃, and performing post-treatment after the reaction is finished, wherein the post-treatment process comprises the following steps: cooling the obtained reaction solution at 0 ℃ for 20min, extracting with deionized water and ethyl acetate, drying the organic phase with anhydrous sodium sulfate, and concentrating under reduced pressure to remove the solvent to obtain an intermediate 2;

the reaction process is as follows:

step S13, mixing chlorinated trimellitic anhydride and pyridine, stirring at the temperature of 0-5 ℃ until white precipitate is separated out, then adding an intermediate 2 and tetrahydrofuran, stirring at room temperature for reaction for 12 hours, and carrying out post-treatment after the reaction is finished, wherein the post-treatment process comprises the following steps: the obtained reaction solution is decompressed, concentrated and the solvent is removed, and then toluene and acetic anhydride are used for recrystallization to obtain the modifier.

The reaction process is as follows:

further, the molar ratio of 2-octenyl succinic anhydride to methanol in step S11 is 1: 1; the using amount of the anhydrous aluminum trichloride is 4 percent of the total mass of the 2-octenyl succinic anhydride and the methanol; the mass fraction of the sulfuric acid solution in the step S12 is 76%, and the dosage ratio of the L-ascorbic acid, the intermediate 1 and the sulfuric acid solution is 2.2 g: 1 g: 5 mL; the use amount ratio of the chlorinated trimellitic anhydride, pyridine, intermediate 2 and tetrahydrofuran in step S13 was 1.2 g: 0.5 g: 2.3 g: 50 mL.

Furthermore, the mesh cloth is one of cotton gauze or non-woven fabric, and the additive is one or more of polyvinylpyrrolidone, gelatin, chitin, chitosan, alginate, polyacrylamide and polyethylene oxide which are mixed according to any proportion.

Further, an application of the ozone hydrogel dressing in wet wound healing.

The invention has the beneficial effects that:

ozone itself has bactericidal effect, can avoid wound infection, and ozone can activate cell metabolism and organism immunity, promote expression of inflammatory factor and growth factor in wound, and promote wound healing. Ozone can activate erythrocyte metabolism, so that immune system and antioxidant system of organism generate slight peroxidation on erythrocyte membrane, improve oxygen transportation and utilization in tissue, increase oxygen supply of important tissue and organ, and improve blood oxygen saturation. In addition, ozone can promote the expression of inflammatory factors and growth factors in wounds.

The agar in the invention is one of hydrophilic colloids, the water loss rate refers to that the polymer network is shrunk and water is separated because the spiral structure is further slowly gathered in the aging process of the gel, carboxyl with stronger hydrophilicity is formed in the agar molecular structure after the agar is modified by a modifier, on the other hand, the formation of the spiral structure is hindered because of the introduction of the hydrophobic hydrocarbon long chain, and the hydroxyl which does not participate in the reaction is exposed outside the spiral structure and combined with water molecules, so that the water loss rate of the modified gel material is obviously reduced.

The hydrogel dressing directly prepares ozonized hydrogel with antibacterial property by reacting ozone with hydrogel matrix, can play roles in reducing the growth of bacteria and preventing wound infection without embedding an antibacterial agent, and can accelerate angiogenesis and promote epithelial cell growth by ozone, thereby promoting wound healing. The hydrogel has good water retention and moisture retention effects, can keep a wound moist, and can stably combine ozone on the hydrogel matrix through a process, so that the ozone plays a role in inhibiting bacteria and promoting the wound healing.

Drawings

In order to facilitate understanding for those skilled in the art, the present invention will be further described with reference to the accompanying drawings.

FIG. 1 is a CT image of the skin under the mouse dermatitis model of example 7;

FIG. 2 is a skin CT image under the dermatitis model in the mouse in comparative example 1;

FIG. 3 is a photograph showing bacteriostatic effects against Staphylococcus aureus in example 7 and comparative example 2;

FIG. 4 is a photograph showing the bacteriostatic effects of example 7 and comparative example 2 on E.coli.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

Preparing a modifier:

step S13, mixing chlorinated trimellitic anhydride and pyridine, stirring at the temperature of 0 ℃ until white precipitate is separated out, then adding an intermediate 2 and tetrahydrofuran, stirring at room temperature for reaction for 12 hours, and carrying out post-treatment after the reaction is finished, wherein the post-treatment process comprises the following steps: the obtained reaction solution is decompressed, concentrated and the solvent is removed, and then toluene and acetic anhydride are used for recrystallization to obtain the modifier.

Wherein the molar ratio of 2-octenyl succinic anhydride to methanol in step S11 is 1: 1; the using amount of the anhydrous aluminum trichloride is 4 percent of the total mass of the 2-octenyl succinic anhydride and the methanol; the mass fraction of the sulfuric acid solution in the step S12 is 76%, and the dosage ratio of the L-ascorbic acid, the intermediate 1 and the sulfuric acid solution is 2.2 g: 1 g: 5 mL; the use amount ratio of the chlorinated trimellitic anhydride, pyridine, intermediate 2 and tetrahydrofuran in step S13 was 1.2 g: 0.5 g: 2.3 g: 50 mL.

Example 2

Preparing a modifier:

step S13, mixing chlorinated trimellitic anhydride and pyridine, stirring at the temperature of 5 ℃ until white precipitate is separated out, then adding an intermediate 2 and tetrahydrofuran, stirring at room temperature for reaction for 12 hours, and carrying out post-treatment after the reaction is finished, wherein the post-treatment process comprises the following steps: the obtained reaction solution is decompressed, concentrated and the solvent is removed, and then toluene and acetic anhydride are used for recrystallization to obtain the modifier.

Example 3

Preparing modified agar:

Wherein the dosage ratio of the agar powder to the deionized water is 3 g: 100mL, and the dosage ratio of the mixed solution of the modifier and the isopropanol is 1 g: 10 mL; the dosage ratio of the agar powder to the mixed liquid of the modifier and the isopropanol is 3 g: 100 mL; the modifier was prepared as in example 2.

Example 4

Preparing modified agar:

Wherein the using amount ratio of the agar powder to the deionized water is 7 g: 100mL, and the dosage ratio of the mixed solution of the modifier and the isopropanol is 1 g: 10 mL; the dosage ratio of the agar powder to the mixed liquid of the modifier and the isopropanol is 2 g: 100 mL; the modifier was prepared as in example 2.

Example 5

Preparing modified agar:

Wherein the dosage ratio of the agar powder to the deionized water is 10 g: 100mL, and the dosage ratio of the mixed solution of the modifier and the isopropanol is 1 g: 10 mL; the dosage ratio of the agar powder to the mixed liquid of the modifier and the isopropanol is 1 g: 100 mL; the modifier was prepared as in example 2.

Example 6

Preparing an ozone hydrogel dressing:

firstly, weighing the following raw materials in parts by weight: 0.5 part of modified agar, 1 part of polyethylene glycol, 2 parts of additive and 60 parts of deionized water;

secondly, mixing the modified agar and 50% of deionized water, heating and stirring the mixture at the temperature of 95 ℃ and the rotating speed of 400r/min until the modified agar is completely dissolved, stopping heating, stirring and cooling the mixture to the temperature of 40 ℃ to obtain a first mixture;

step three, mixing the additive and polyethylene glycol uniformly, then adding the rest deionized water, stirring and mixing at the rotation speed of 400r/min to obtain a second mixture, adding the second mixture into the first mixture, introducing ozone while stirring, controlling the system temperature at 25 ℃, the pressure at 1.5bar and the ozone content at 80g/L, then pouring into a culture dish with mesh cloth, and carrying out gamma-ray irradiation, wherein the irradiation dose is as follows: 30Gy, and irradiating for 2h to obtain the ozone hydrogel dressing.

Wherein the mesh cloth is one of cotton gauze or non-woven fabric; the additive is a mixture of polyvinylpyrrolidone, chitin, chitosan, alginate, polyacrylamide and polyethylene oxide which are mixed according to equal mass; modified agar was prepared as in example 4.

Example 7

Preparing an ozone hydrogel dressing:

firstly, weighing the following raw materials in parts by weight: 1.5 parts of modified agar, 10 parts of polyethylene glycol, 10 parts of additive and 70 parts of deionized water;

and thirdly, mixing the additive and polyethylene glycol uniformly, adding the rest deionized water, stirring and mixing at the rotation speed of 400r/min to obtain a second mixture, adding the second mixture into the first mixture, introducing ozone while stirring, controlling the temperature of the system at 30 ℃, the pressure at 1bar and the ozone content at 100g/L, then pouring the mixture into a culture dish with gridding cloth, and carrying out gamma-ray irradiation with the irradiation dose of 60kGy for 1h to obtain the ozone water gel dressing.

Example 8

Preparing an ozone hydrogel dressing:

firstly, weighing the following raw materials in parts by weight: 2 parts of modified agar, 20 parts of polyethylene glycol, 20 parts of an additive and 85 parts of deionized water;

secondly, mixing the modified agar and 50% of deionized water, heating and stirring the mixture at the temperature of 100 ℃ and the rotating speed of 500r/min until the modified agar is completely dissolved, stopping heating, stirring and cooling the mixture to the temperature of 40 ℃ to obtain a first mixture;

step three, mixing the additive and polyethylene glycol uniformly, then adding the rest deionized water, stirring and mixing at the rotation speed of 500r/min to obtain a second mixture, adding the second mixture into the first mixture, introducing ozone while stirring, controlling the system temperature at 35 ℃, the pressure at 1.5bar, the ozone content at 120g/L, then pouring into a culture dish with mesh cloth, carrying out gamma-ray irradiation, and irradiating with the dose: 90kGy, and the irradiation time is 0.5h, thus obtaining the ozone hydrogel dressing.

Comparative example 1

The modified agar from example 7 was replaced with unmodified agar, and the remaining raw materials and preparation process were kept unchanged.

Comparative example 2

Compared with the example 7, ozone is not introduced, and the rest raw materials and the preparation process are kept unchanged.

The samples prepared in examples 6 to 8 and comparative example 1 were tested:

1) cutting the sample into 2 x 2cm, drying for 64h at the temperature of 30 ℃, and testing the water loss rate;

2) samples were cut into 2 x 2cm and then placed in deionized water and normal saline, respectively, to test for weight gain.

The test results are shown in table 1 below:

TABLE 1

Item	Example 6	Example 7	Example 8	Comparative example 1
					Water loss (%)	65.2	65.1	65.3	80.2
Water absorption weight gain (g)	1.07	1.10	1.08	0.93
					Weight gain (g) by absorption of physiological saline	0.82	0.82	0.83	0.69

Ozonized gels with different concentrations are prepared according to different ozone introducing time, the higher the ozone concentration is, the better the antibacterial effect is, and the performance of the gel is evaluated through tests such as drying, imbibition, morphological observation and the like, and the gel dressing prepared by the method disclosed by the invention is better in water retention effect and more stable as can be seen from table 1.

Samples were prepared for examples 6-8, and the pH of the hydrogels were measured using a pH meter, respectively, at a pH range of 4.5-6.0, which is compatible with the normal skin pH of humans.

The samples prepared in example 7 and comparative example 1 were subjected to drug absorption testing:

wherein the drug is timolol;

the timolol is found to have stronger absorption peaks at 295nm and 205nm by advanced wavelength scanning, but an interference peak appears at 205nm, so 295nm is selected as the absorption wavelength of timolol, a standard curve is established first, then the absorbance values before and after soaking are measured, and finally the absorption quantity of the hydrogel to timolol is calculated, and the result is shown in the following table 2:

TABLE 2

As can be seen from Table 2, the hydrogel samples prepared by the present invention have good drug absorption.

It is seen from the water loss rate that the hydrogel state provides a large amount of moisture to the skin, thereby maintaining the proper moisturization of the wound and normal skin. It is seen from the swelling ratio that in the dry state, the gel can absorb a large amount of water, so that the dressing can rapidly absorb wound exudate, thereby preventing wound immersion and microorganism breeding. Thus having a two-way regulating effect. As a drug carrier, the load of timolol under the unit area of 2cm by 2cm reaches 10.79mg, which indicates that the hydrogel can be used as a good drug carrier.

The samples of example 7 and comparative example 1 were subjected to clinical use in a mouse dermatitis model, and the effects of use are shown in fig. 1-2:

sample of example 7: the number of inflammatory cells was greatly reduced, as shown in figure 1;

sample of comparative example 1: the expansion of inflammatory cells and blood vessels was still evident after the dermatitis treatment, and the change was less obvious than before the treatment, as shown in fig. 2.

The samples prepared in example 7 and comparative example 2 were subjected to a bacteriostasis test:

the bacteriostatic effects of example 7 and comparative example 2 on staphylococcus aureus and escherichia coli are examined by referring to a bacteriostatic test method in the 2002 edition of disinfection technical specification;

the photos of the bacteriostatic effect of example 7 and comparative example 2 on staphylococcus aureus are shown in fig. 3, wherein the left culture medium in fig. 3 is comparative example 2, and the right culture medium is example 7; the photographs of the bacteriostatic effects of example 7 and comparative example 2 on E.coli are shown in FIG. 4, the left side medium is comparative example 2, and the right side medium is example 7.

The results of the bacteriostasis rate are as follows: the calculation formula is X ═ (A-B)/A × 100%;

in the formula: x represents the bacteriostasis rate; a is the average colony number of comparative example 2; b is the average colony number of example 7; the results of the bacteriostatic rate are shown in table 2 below:

TABLE 2

Bacterial strain	Bacteriostatic ratio (%)
		Staphylococcus aureus	81.4
Escherichia coli	82.4

As can be seen from the above table 2, the hydrogel prepared by the method has 81.4% and 82.4% of bacteriostatic rates on Staphylococcus aureus and Escherichia coli, respectively, and has an obvious inhibitory effect.

In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention.

In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. The foregoing is illustrative and explanatory only and is not intended to be exhaustive or to limit the invention to the precise embodiments described, and various modifications, additions, and substitutions may be made by those skilled in the art without departing from the scope of the invention or exceeding the scope of the claims.

Claims

1. The preparation method of the ozone hydrogel dressing is characterized by comprising the following steps of:

secondly, mixing the modified agar and 50% of deionized water, heating and stirring at the temperature of 95-100 ℃ until the modified agar is completely dissolved, stopping heating, stirring and cooling to the temperature of 40 ℃ to obtain a first mixture;

thirdly, mixing the additive and the polyethylene glycol uniformly, adding the rest deionized water, stirring to obtain a second mixture, adding the second mixture into the first mixture, introducing ozone while stirring, then pouring into a culture dish with grid cloth, and performing gamma-ray irradiation to obtain an ozone water gel dressing;

the modified agar is prepared by the following steps:

mixing agar powder and deionized water, stirring at the temperature of 35 ℃, adjusting the pH value to 9 by using 1mol/L sodium hydroxide solution, then adding a mixed solution of a modifier and isopropanol, dropwise adding the mixed solution within 1.5h, keeping the temperature unchanged after dropwise adding, continuing stirring for 3h, adjusting the pH value to 6 by using 1mol/L hydrochloric acid solution, and performing post-treatment after reaction to obtain the modified agar.

2. The method for preparing the ozone hydrogel dressing according to claim 1, wherein when ozone is introduced, the temperature of the system is controlled to be 25-35 ℃, the pressure is 0.5-1.5bar, and the ozone content reaches 80-120 g/L; when gamma ray irradiation is performed, irradiation dose: 30-90kGy and irradiation time of 0.5-2 h.

3. The method for preparing the ozone hydrogel dressing according to claim 1, wherein the ratio of the agar powder to the deionized water is 3-10 g: 100mL, and the dosage ratio of the mixed solution of the modifier and the isopropanol is 1 g: 10 mL; the dosage ratio of the agar powder to the mixed liquid of the modifier and the isopropanol is 1-3 g: 100 mL.

4. The method for preparing an ozone hydrogel dressing according to claim 1, wherein the modifier is prepared by the following steps:

step S11, mixing 2-octenyl succinic anhydride and methanol, stirring to be transparent at the temperature of 45 ℃, then heating to 60 ℃, reacting for 2 hours, then adding anhydrous aluminum trichloride, reacting for 2.5 hours at the temperature of 80 ℃, and performing post-treatment after the reaction is finished to obtain an intermediate 1;

step S12, dripping L-ascorbic acid and the intermediate 1 into a sulfuric acid solution, stirring for 1h at the temperature of 20 ℃, reacting for 28h at the temperature of 25 ℃, and performing post-treatment after the reaction is finished to obtain an intermediate 2;

and step S13, mixing chlorinated trimellitic anhydride and pyridine, stirring at the temperature of 0-5 ℃ until white precipitate is separated out, then adding the intermediate 2 and tetrahydrofuran, stirring at room temperature for reaction for 12 hours, and carrying out post-treatment after the reaction is finished to obtain the modifier.

5. The method for preparing the gel dressing with ozone water as claimed in claim 4, wherein the molar ratio of 2-octenyl succinic anhydride to methanol in step S11 is 1: 1; the using amount of the anhydrous aluminum trichloride is 4 percent of the total mass of the 2-octenyl succinic anhydride and the methanol; the mass fraction of the sulfuric acid solution in the step S12 is 76%, and the dosage ratio of the L-ascorbic acid, the intermediate 1 and the sulfuric acid solution is 2.2 g: 1 g: 5 mL; the use amount ratio of the chlorinated trimellitic anhydride, pyridine, intermediate 2 and tetrahydrofuran in step S13 was 1.2 g: 0.5 g: 2.3 g: 50 mL.

6. The method for preparing the gel dressing with ozone water as claimed in claim 1, wherein the mesh cloth is one of cotton gauze or non-woven cloth, and the additive is one or more of polyvinylpyrrolidone, gelatin, chitin, chitosan, alginate, polyacrylamide and polyethylene oxide mixed at any ratio.

7. Use of an ozonated water gel dressing prepared according to the process of claim 1 in wet wound healing.