阅读说明：本技术 一种缓释抗菌的敷料及其制备方法 (Slow-release antibacterial dressing and preparation method thereof ) 是由 黄洁君 何珊 李志清 万水停 黄少君 梁丽菁 许雷 黄宇宙 黄邦煜 黄佳裕 于 2021-09-28 设计创作，主要内容包括：本发明属于医药技术领域,公开了一种缓释抗菌的敷料及其制备方法。该敷料包括载体、抗菌物质、穿心莲提取物、凝胶物质。该敷料,将抗菌物质包埋在穿心莲提取物和凝胶物质形成的凝胶状混合物中,形成的纳米级悬液渗入敷料的载体中,从而使得敷料中的抗菌物质具有缓释效果,对人体安全无毒性。该敷料具有广谱抗菌,高渗透性,且对耐药菌株耐甲氧西林金黄色葡萄球菌有良好杀菌作用。(The invention belongs to the technical field of medicines, and discloses a slow-release antibacterial dressing and a preparation method thereof. The dressing comprises carrier, antibacterial substance, herba Andrographitis extract, and gel substance. According to the dressing, the antibacterial substance is embedded in the gel-like mixture formed by the andrographis paniculata extract and the gel substance, and the formed nano-scale suspension permeates into the carrier of the dressing, so that the antibacterial substance in the dressing has a slow-release effect and is safe and non-toxic to a human body. The dressing has broad-spectrum antibacterial and high permeability, and has good bactericidal effect on drug-resistant strains such as methicillin-resistant staphylococcus aureus.)

1. A dressing is characterized by comprising a carrier, an antibacterial substance, an andrographis paniculata extract and a gel substance.

2. The dressing of claim 1, wherein the antimicrobial substance comprises Australian tea tree essential oil.

3. The dressing of claim 1, wherein the dressing further comprises an emulsifier.

4. A dressing according to claim 1 or 3, wherein the dressing further comprises water.

5. The dressing of claim 1, wherein the dressing comprises 0.01-25 parts by weight of an antibacterial substance, 0.1-30 parts by weight of an andrographis paniculata extract, and 0.1-10 parts by weight of a gel substance.

6. The dressing of claim 3, wherein the dressing comprises 0.01-25 parts by weight of antibacterial substance, 0.1-5 parts by weight of emulsifier, 0.1-30 parts by weight of andrographis paniculata extract, and 0.1-10 parts by weight of gel substance.

7. The dressing of claim 4, consisting of the following components in parts by weight: 5-17 parts of antibacterial substance, 1-3.5 parts of emulsifier, 7.5-20 parts of common andrographis herb extract, 2.5-7 parts of gel substance and the balance of water.

8. A method of manufacturing a dressing according to any one of claims 1 to 7, comprising the steps of:

(1) stirring and mixing the andrographis paniculata extract and the gel substance to obtain a gel-like mixture;

(2) in the process of stirring the gelatinous mixture, dripping the antibacterial substance into the gelatinous mixture, and homogenizing to prepare nano-scale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, and taking out the carrier to prepare the dressing.

9. The production method according to claim 8, wherein the antibacterial substance is mixed with an emulsifier before dropping; in the step (1), the andrographis paniculata extract, the gel substance and water are stirred and mixed.

10. Use of a dressing according to any one of claims 1 to 7 in the manufacture of a medicament.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a slow-release antibacterial dressing and a preparation method thereof.

Background

The skin is the largest organ of the human body and is the first barrier of the human body, but the skin and mucous membrane are inevitably damaged in daily work and life. Since the UK zoologist Winter provided the concept of 'moist healing' in the last 60 years, the traditional healing concept of self-scabbing and drying wounds cannot meet the requirement of people on recovery of skin injury due to long healing period, easy bacterial breeding and inconvenience for wound healing. Many wet medical dressings are in transit. Although the existing dressings are various in types and can meet the healing requirements of most wound surfaces, the antibacterial effect is still poor, particularly, the antibacterial types are limited, the dressings need to be frequently replaced, the large-area deep wound surfaces can be subjected to fungal and drug-resistant bacteria double infection after being applied with external medicines and dressings, and the dressings need to be frequently replaced.

Therefore, a new dressing with excellent antibacterial effect, especially against drug-resistant bacteria, is needed.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the dressing has the slow-release antibacterial effect, not only has broad-spectrum antibacterial effect and good antibacterial effect, but also has good antibacterial effect on methicillin-resistant staphylococcus aureus (MRSA), so that the skin wound can be quickly healed, and the scar of the healed wound is not obvious.

The invention conception of the invention is as follows: according to the antibacterial dressing provided by the invention, the antibacterial substance is embedded in the gel mixture formed by the andrographis paniculata extract and the gel substance, and the formed nano-scale suspension permeates into the carrier of the dressing, so that the antibacterial substance in the dressing has a slow-release effect (the preparation process of the dressing can be called as the preparation of the dressing by adopting a nano-embedding technology), the water solubility of the indissolvable antibacterial active ingredient can be obviously improved, the transdermal absorption of the antibacterial active ingredient is promoted, the stability of the antibacterial active ingredient is improved, the irritation and the toxic and side effects of the antibacterial active ingredient are reduced, the bioavailability of the antibacterial active ingredient is obviously improved, and the antibacterial active ingredient is various. In addition, the dressing has good degradability and good antibacterial effect on methicillin-resistant staphylococcus aureus (MRSA).

In a first aspect of the invention, a slow release antimicrobial dressing is provided.

Specifically, the slow-release antibacterial dressing comprises a carrier, an antibacterial substance, an andrographis paniculata extract and a gel substance.

Preferably, the antibacterial substance comprises Australian tea tree essential oil. The Australian tea tree essential oil plant active component contains 4-terpineol and 1, 8-cineole, and the Australian tea tree essential oil is an excellent natural antibacterial agent, has a good killing effect on pathogenic bacteria on the surface of human skin, and has the functions of resisting infection, enhancing the body immunoregulation capability and accelerating wound healing.

Preferably, the dressing further comprises an emulsifier. The mixed Australian tea tree essential oil and emulsifier can improve the stability of the antibacterial active ingredients in the antibacterial substance, thereby slowly releasing the antibacterial active ingredients.

Preferably, in the Australian tea tree essential oil, the content of 4-terpineol is 35-48% by mass, and the content of 1, 8-cineole is not more than 4.5%.

Preferably, the Australian tea tree essential oil further comprises at least one of alpha-pinene, sabinene, alpha-terpinene, limonene, p-cymene, gamma-terpinene, terpinolene, myrcene, allene, delta-cadinene, eudesmol and melaleucinol.

Further preferably, the emulsifier is at least one selected from tween 80, span 80, alkyl glycoside, PEG-40 hydrogenated castor oil (polyoxyethylene ether-40 castor oil), glyceryl stearyl phosphate, hydrogenated lecithin, fatty alcohol polyoxyethylene ether, and polysorbate-80.

Preferably, the gel substance comprises at least one of polyacrylamide dimethyl taurinate, carbomer, triethanolamine, chitosan derivative, gelatin, acrylic acid/C10-30 alkanol acrylate cross-linked polymer.

Preferably, the carrier is selected from at least one of cotton cloth or aged/oxidized cotton cloth, non-woven fabric, gauze, terylene and porous organic polymer material. The length and width of the carrier specification can be 10 x 15cm, 20 x 25cm and 40 x 50cm, and the carrier needs to have the porous characteristic, so that the dressing has good air permeability.

Preferably, the dressing further comprises water.

Preferably, the dressing comprises 0.01-25 parts of antibacterial substances, 0.1-30 parts of common andrographis herb extracts and 0.1-10 parts of gel substances by weight.

Preferably, the dressing comprises, by weight, 5-17 parts of an antibacterial substance, 7.5-20 parts of an andrographis paniculata extract and 2.5-7 parts of a gel substance.

Preferably, the dressing comprises, by weight, 0.01-25 parts of an antibacterial substance, 0.1-5 parts of an emulsifier, 0.1-30 parts of an andrographis paniculata extract, and 0.1-10 parts of a gel substance.

Preferably, the dressing comprises, by weight, 5-17 parts of an antibacterial substance, 1-3.5 parts of an emulsifier, 7.5-20 parts of an andrographis paniculata extract, and 2.5-7 parts of a gel substance.

More preferably, the dressing consists of the following components in parts by weight: 5-17 parts of antibacterial substance, 1-3.5 parts of emulsifier, 7.5-20 parts of common andrographis herb extract, 2.5-7 parts of gel substance and the balance of water.

In a second aspect, the invention provides a method for preparing a slow-release antibacterial dressing.

Specifically, the preparation method of the slow-release antibacterial dressing comprises the following steps:

(1) mixing herba Andrographitis extract and gel material under stirring to obtain gel mixture;

(2) in the process of stirring the gelatinous mixture, dripping the antibacterial substance into the gelatinous mixture, and homogenizing to prepare nano-scale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, and taking out the carrier to prepare the dressing.

Preferably, the antibacterial substance is mixed with the emulsifier before dropping, preferably by ultrasonic treatment.

Preferably, in the step (1), the andrographis paniculata extract, the gel substance and water are mixed with stirring.

Preferably, in the step (1), the stirring speed of the stirring and mixing is 500-1200 rpm; further preferably, the stirring speed of the stirring and mixing is 500-1100 rpm. Preferably, a magnetic stirrer is used for stirring.

Preferably, in the step (2), the homogenization process is carried out for 1.5-5 minutes by using a high-pressure homogenizer, and the homogenization speed is 8800-9000 rpm.

Preferably, in the step (3), when the dressing is taken out, a saturated but non-dripping state is formed between the nano-scale suspension and the carrier.

A third aspect of the invention provides a use comprising a dressing as described above.

In particular to the application of the dressing in preparing medicines.

Further preferably, the dressing is applied to the preparation of antibacterial drugs.

More preferably, the dressing is applied to the preparation of a medicament for treating skin injury.

The slow release effect of the dressing can reach 24 hours, and the dressing has a good antibacterial effect within 24 hours.

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

(1) according to the antibacterial dressing provided by the invention, the antibacterial substance is embedded in the gel mixture formed by the andrographis paniculata extract and the gel substance, and the formed nano-scale suspension permeates into the carrier of the dressing, so that the antibacterial substance in the dressing has a slow release effect, has a good antibacterial effect within 24 hours, is safe and nontoxic to human bodies, and is an ideal wound external dressing.

(2) The dressing contains Australia tea tree essential oil, is used as an antibacterial substance, and has broad-spectrum antibacterial property and high permeability. Can simultaneously resist three microorganisms, namely bacteria, fungi and viruses, and has good bactericidal action on drug-resistant strains, namely methicillin-resistant staphylococcus aureus (MRSA).

(3) The dressing has the advantages of good air permeability, good biocompatibility, strong affinity, fast degradation and contribution to the renewal, metabolism and healing of skin wound tissue cells.

Detailed Description

In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.

The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.

The Australian tea Tree essential oil used in the following examples is also known as MELALEUCA ALTERNIFOLIA (Melaleuca ALTERNIFOLIA) leaf oil, and is available from Nanning Wanjiahui perfumery, Inc.

The gel material (carbomer 941) used in the following examples is available from shanghai spectral vibration biotechnology limited under lot number: ZY 210907.

Example 1: preparation of the dressing

A slow-release antibacterial dressing comprises, by weight, a carrier (cotton cloth), 0.01 part of Australian tea tree essential oil, 0.01 part of an emulsifier (Tween 80), 30 parts of an andrographis paniculata extract, 0.1 part of a gel substance (polyacrylamide dimethyl taurinate), and 69.88 parts of water.

The preparation method of the slow-release antibacterial dressing comprises the following steps:

(1) mixing herba Andrographitis extract, gel material and water under stirring at 1000 rpm to obtain gel mixture;

(2) in the process of stirring the gel-like mixture, carrying out ultrasonic treatment on an antibacterial substance and an emulsifier for mixing, dripping the obtained antibacterial mixture into the gel-like mixture, continuing stirring for 10 minutes, and homogenizing for 3 minutes by using a high-pressure homogenizer at the homogenizing speed of 9000 r/min to obtain a nanoscale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, taking out the carrier, and forming a saturated but non-dripping state between the nano-scale suspension and the carrier when taking out the dressing to obtain the dressing.

Example 2: preparation of the dressing

A slow-release antibacterial dressing comprises, by weight, a carrier (cotton cloth), 5 parts of Australian tea tree essential oil, 1 part of an emulsifier (Tween 80, span 80, 0.5 part of each), 20 parts of an andrographis paniculata extract, 10 parts of a gel substance (carbomer 941 and triethanolamine, 9 parts and 1 part respectively) and 64 parts of water.

The preparation method of the slow-release antibacterial dressing comprises the following steps:

(1) mixing herba Andrographitis extract, gel material and water under stirring at 1000 rpm to obtain gel mixture;

(2) in the process of stirring the gel-like mixture, carrying out ultrasonic treatment on an antibacterial substance and an emulsifier for mixing, dripping the obtained antibacterial mixture into the gel-like mixture, continuing stirring for 10 minutes, and homogenizing for 3 minutes by using a high-pressure homogenizer at the homogenizing speed of 9000 r/min to obtain a nanoscale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, taking out the carrier, and forming a saturated but non-dripping state between the nano-scale suspension and the carrier when taking out the dressing to obtain the dressing.

Example 3: preparation of the dressing

A slow-release antibacterial dressing comprises, by weight, a carrier (cotton cloth), 12.5 parts of Australian tea tree essential oil, 3.5 parts of an emulsifier (polysorbate-80), 15 parts of an andrographis paniculata extract, 5 parts of a gel substance (ammonium polyacryloyldimethyltaurate) and 64 parts of water.

The preparation method of the slow-release antibacterial dressing comprises the following steps:

(1) mixing herba Andrographitis extract, gel material and water under stirring at 1000 rpm to obtain gel mixture;

(2) in the process of stirring the gel-like mixture, carrying out ultrasonic treatment on an antibacterial substance and an emulsifier for mixing, dripping the obtained antibacterial mixture into the gel-like mixture, continuing stirring for 10 minutes, and homogenizing for 3 minutes by using a high-pressure homogenizer at the homogenizing speed of 9000 r/min to obtain a nanoscale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, taking out the carrier, and forming a saturated but non-dripping state between the nano-scale suspension and the carrier when taking out the dressing to obtain the dressing.

Example 4: preparation of the dressing

A slow-release antibacterial dressing comprises, by weight, a carrier (cotton cloth), 17 parts of Australian tea tree essential oil, 2.5 parts of emulsifiers (Tween 80 and span 80, 1.25 parts of each), 10 parts of an andrographis paniculata extract, 2.5 parts of a gel substance (polyacrylamide dimethyl taurinate) and 68 parts of water.

The preparation method of the slow-release antibacterial dressing comprises the following steps:

(1) mixing herba Andrographitis extract, gel material and water under stirring at 1000 rpm to obtain gel mixture;

(2) in the process of stirring the gel-like mixture, carrying out ultrasonic treatment on an antibacterial substance and an emulsifier for mixing, dripping the obtained antibacterial mixture into the gel-like mixture, continuing stirring for 10 minutes, and homogenizing for 3 minutes by using a high-pressure homogenizer at the homogenizing speed of 9000 r/min to obtain a nanoscale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, taking out the carrier, and forming a saturated but non-dripping state between the nano-scale suspension and the carrier when taking out the dressing to obtain the dressing.

Example 5: preparation of the dressing

A slow-release antibacterial dressing comprises, by weight, a carrier (cotton cloth), 25 parts of Australian tea tree essential oil, 5 parts of emulsifiers (Tween 80 and span 80, 2.5 parts of each), 0.1 part of an andrographis paniculata extract, 7 parts of a gel substance (polyacrylamide dimethyl taurinate) and 62.9 parts of water.

The preparation method of the slow-release antibacterial dressing comprises the following steps:

(1) mixing herba Andrographitis extract, gel material and water under stirring at 1000 rpm to obtain gel mixture;

(2) in the process of stirring the gel-like mixture, carrying out ultrasonic treatment on an antibacterial substance and an emulsifier for mixing, dripping the obtained antibacterial mixture into the gel-like mixture, continuing stirring for 15 minutes, and homogenizing for 5 minutes by using a high-pressure homogenizer at the homogenizing speed of 9000 r/min to obtain a nanoscale suspension;

(3) and (3) immersing the carrier into the nano-scale suspension, taking out the carrier, and forming a saturated but non-dripping state between the nano-scale suspension and the carrier when taking out the dressing to obtain the dressing.

Comparative example 1

In comparison with example 3, the dressing of comparative example 1 did not contain a gel substance, and the rest of the composition and the preparation process were the same as example 3. I.e. the dressing of comparative example 1 was prepared without the addition of a gel substance.

Comparative example 2

In comparison with example 3, the dressing of comparative example 2 does not contain Andrographis paniculata Nees extract, and the rest of the composition and preparation process are the same as example 3. That is, the dressing of comparative example 2 was prepared without the addition of Andrographis paniculata Nees extract.

Product Effect test (for convenience of the following experiment, the effect of the present invention using the nano-sized suspension prepared in step (2) of the example of the present invention, which represents the effect of the dressing, was performed using the nano-sized suspension prepared in step (2) of the present invention instead of the dressing)

1. Test for efficacy against methicillin-resistant Staphylococcus aureus (MRSA)

The operation steps of the test for soaking and quantitatively sterilizing the carrier according to the disinfection technical specification 2002 edition are as follows:

1) several sterile plates were used, and it was indicated that the nanosuspensions for preparing dressings prepared in examples 1 to 5 and comparative examples 1 to 2 were injected into the plates in an amount of 5.0mL per plate.

2) After the plate containing the nano-sized suspensions prepared in examples 3 to 5 and comparative examples 1 to 2 was placed in a water bath tank at 20 ℃. + -. 1 ℃ for 5 minutes, 3 pieces of methicillin-resistant Staphylococcus aureus (MRSA) slides prepared in advance were placed in each of the plates using sterile tweezers and were allowed to soak in the nano-sized suspensions (as test groups) prepared in examples 3 to 5 and comparative examples 1 to 2.

3) After the methicillin-resistant staphylococcus aureus and the nano-scale suspension interact for 1 hour at the preset time, the bacterial tablets are taken out by using sterile forceps and respectively transferred into test tubes containing 5.0mL of neutralizing agent (2% of Tween-80, 0.5% of lecithin with mass concentration, 1% of glycine with mass concentration and 0.03mol/L of PBS phosphate buffered saline solution). Mixing with electric mixer for 20s, or shaking the test tube on palm for 80 times to elute bacteria on the bacterial sheet into the neutralizing solution, and standing for more than 5 min to achieve sufficient neutralizing effect. Finally, after further mixing, 1.0mL of the solution was pipetted and inoculated directly to 2 plates per tube, and the number of viable bacteria was determined.

4) Another dish was filled with 10.0mL of sterile hard water, and 3 discs of mushroom were placed as a positive control. The subsequent test procedure and viable bacteria culture count were the same as in the test group described above.

5) Culturing all test samples in an incubator at 37 ℃, and culturing the bacterial propagules for 48 hours to observe the final result; the bacterial spores were cultured for 72 hours to observe the final results.

6) The above test group and control group were repeated 3 times, and the viable bacterial count (cfu/tablet) of each group was calculated.

7) Calculating the killing rate:

the killing rate (%) is N_t＝(V₀-V_t)/V₀×100％；

Nt is the killing rate of bacteria in the experiment;

V₀and V_tRespectively sterilizing the positive control groupBefore and after sterilization (V before sterilization)₀After sterilization is V_t) The bacteria content.

Results of the antibacterial experiments before and after using the nano-sized suspension are shown in table 1.

Table 1: antibacterial results

As can be seen from table 1, the dressings prepared in examples 3 to 5 of the present invention have an obvious inhibitory effect on methicillin-resistant staphylococcus aureus (MRSA) and a better antibacterial activity against drug-resistant bacteria, wherein the dressing prepared in example 3 is the best for methicillin-resistant staphylococcus aureus (MRSA).

2. Broad spectrum antibacterial test

Taking the colonies of escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and candida albicans cultured for 18-24 hours, and preparing bacterial suspensions (called test bacterial suspensions) meeting the experimental requirements by respectively adopting a direct bacterial suspension preparation method.

1mL of test bacterial suspension and 4mL of the nanosuspension prepared in example 3 were added to a sterile tube, mixed, and incubated (20. + -. 1 ℃) for 5 minutes, 10 minutes, and 20 minutes. Then 0.5mL of the sample solution in the sterile test tube was added to a test tube containing 4.5mL of a neutralizing agent (2% by mass of Tween 80, 0.5% by mass of lecithin, 1% by mass of glycine, and 0.03mol/L of PBS phosphate buffered saline) and mixed uniformly, and the mixture was neutralized for 10 minutes. Respectively sucking 1.0mL of sample liquid, measuring the number of viable bacteria according to a viable bacteria culture counting method, and calculating the killing rate. If the number of colonies growing on the plate is large, serial 10-fold dilution can be performed, and viable bacteria culture counting can be performed. In parallel, a dilution (a reagent for field test) was used instead of the mixture of example 3, and a positive control group was prepared. The results are shown in Table 2.

Table 2: test results of broad-spectrum antibacterial property

As can be seen from Table 2, the dressing prepared in example 3 has a significant effect on killing Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans.

3. Actual irritation test on damaged skin

Taking 24 SD rats (SD rat is a strain of rat) with the size, the weight of 160-. SD rats were randomly divided into 8 groups of 3 rats each (3 SD rats No. 1, 2, 3). The hair on the back of each SD rat was shaved with an electric razor, and the area of the shaved hair on the back was approximately 14cm²1 burn wound with uniform size, area range, depth and severity is prepared according to the proportion of 2cm²Skin dose of 500. mu.L was administered to each group of SD rats separately, each SD rat was drawn at 12cm²That is, 3mL of the drug (nano-sized suspension for dressing prepared in examples 1-5 and comparative examples 1-2, respectively, and a blank control group was administered with physiological saline) was administered to each SD rat, fixed by application, and observed after 24 hours, 48 hours, and 72 hours. The evaluation criteria for specific skin irritation are shown in tables 3 and 4, and the results of observation are shown in table 5.

TABLE 3

TABLE 4

TABLE 5

The fractional scores given for erythema and edema in Table 5 are scored with reference to the scores given in Table 3.

As can be seen from Table 5, the nanosuspensions prepared in examples 1-5 of the present invention for preparing dressings exhibited no erythema or edema for 24 hours, 48 hours, and 72 hours. The nano-scale suspension used for preparing the dressing in the embodiment 3 of the invention has the best effect, so that the dressing has low irritation and good slow-release effect.