A probiotic antibacterial gel dressing and preparation method thereof
By encapsulating E. coli Nissle 1917 and Poria cocos extract in gel dressings with polyvinyl alcohol and moisturizer, the problems of low resistance and antibacterial persistence of antibacterial gel dressings were solved, and efficient antibacterial effect and wound healing were achieved Promote.
Patent Information
- Application Number
- CN202410391538.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-04-01
- Publication Date
- 2024-12-13
- Estimated Expiration
- 2044-04-01
AI Technical Summary
Existing antibacterial gel dressings are prone to lead to drug resistance and cumulative toxicity during long-term use, and have low antibacterial durability, making it difficult to effectively inhibit the growth of drug-resistant bacteria such as Pseudomonas aeruginosa and affect wound healing.
Antibacterial gel dressings prepared with probiotics such as E. coli Nissle 1917 and Poria extract, encapsulate probiotics through a combination of polyvinyl alcohol, moisturizer and ultrapure water to improve their colonization and survival rate, and through polysaccharides, The triterpenes, fatty acids, sterols and other ingredients enhance the antibacterial effect.
It realizes the durability and low toxicity of antibacterial ability, avoids the occurrence of drug resistance, significantly improves the colonization rate and survival rate of probiotics in wounds, promotes wound healing, and has good moisturizing and film forming properties. It is suitable for Treatment of wound and catheter infection.
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of antibacterial dressings, and in particular to a probiotic antibacterial gel dressing and a preparation method thereof. Background Art
[0002] At present, due to the abuse of antibiotics, some pathogenic bacteria have developed resistance to antibiotics. Wound infection caused by resistant bacteria is a huge challenge in clinical treatment. In particular, Pseudomonas aeruginosa infection causes severe inflammatory response at the wound site, prolongs wound healing time, and may even form unhealable wounds, leading to high mortality in patients. Therefore, it is urgent to develop "non-antibiotic" strategies to combat related infections caused by resistant bacteria while promoting wound healing bioactive wound dressings.
[0003] Studies have shown that probiotics have good antibacterial, immunomodulatory and pathogen-induced microbial dysbiosis effects, which are beneficial to wound repair and tissue repair. Among them, Escherichia coli Nissle 1917 (EcN) effectively inhibits the growth of pathogenic bacteria by releasing antimicrobial peptides, iron ion carriers, enzymes and other substances. EcN can inhibit the colonization of pathogenic bacteria (such as enterohemorrhagic Escherichia coli) and the production of toxins (such as Shiga toxin-producing Escherichia coli). In addition, ECN regulates inflammation by affecting the expression of miRNA in tissues. As natural active substances, triterpenoids and polysaccharides in Poria cocos extract have a variety of physiological activities, such as improving the body's immunity, antibacterial, hypoglycemic, and anti-tumor effects. However, the two are affected by external factors during local application on the skin, and their colonization rate and survival rate at the site of injury are low. How to improve their colonization efficiency and survival activity is a problem to be solved.
[0004] The structure of the gel dressing can simulate the cell matrix, and has the advantages of high encapsulation efficiency, controllable porosity, continuous drug release, excellent mechanical properties, and good structural stability. Encapsulating probiotics in the gel dressing can increase the colonization rate and survival rate of probiotics in the wound. Using it as a wound dressing to treat infected wounds has important clinical application value. Summary of the invention
[0005] The purpose of the present invention is to provide a probiotic antibacterial gel dressing and a preparation method thereof, which has the advantages of lasting antibacterial ability, low toxicity, no drug resistance, and no side effects. The dressing can be applied to wound dressings, catheter infections, gastrointestinal infections and other fields, and can solve the problems of drug resistance, cumulative toxicity, and low antibacterial persistence caused by long-term use of antibacterial gel dressings in the prior art.
[0006] To achieve the above-mentioned purpose, the present invention provides a probiotic antibacterial gel dressing, comprising the following components by weight: 5% to 25% of polyvinyl alcohol, 0.5% to 5% of engineered probiotics, 5% to 20% of moisturizer, and the balance is ultrapure water; wherein the engineered probiotics include Poria cocos extract, dopamine and Escherichia coli, the effective ingredients of the Poria cocos extract include polysaccharides, triterpenes, fatty acids, sterols, and enzymes, and the Escherichia coli is Escherichia coli Nissle 1917.
[0007] Furthermore, the composition comprises the following components by weight: 10% to 20% polyvinyl alcohol, 2% to 5% engineered probiotics, 12% to 15% moisturizer, and the balance is ultrapure water.
[0008] Furthermore, the moisturizing agent is one or more of 1,3-propylene glycol, glycerol, 1,3-butylene glycol, sorbitol, and polyethylene glycol.
[0009] Furthermore, the preparation method of the engineered probiotics is as follows:
[0010] S1. Centrifuge the E. coli cultured overnight in LB liquid medium and resuspend it in PBS;
[0011] S2, repeat step S1 three times, and resuspend the E. coli in 10 mM Tirs-HCl at pH 8.5 for the third time;
[0012] S3. Add 50 mg / mL dopamine hydrochloride to the E. coli solution and stir for 30 min;
[0013] S4, adding 100 mg / mL Poria extract to the solution of step S3, and stirring for 30 min;
[0014] S5. Centrifuge the system in step S4 and resuspend it in ice PBS to obtain engineered probiotics.
[0015] The present invention also provides a method for preparing the above-mentioned probiotic antibacterial gel dressing, comprising the following steps:
[0016] a. Mix polyvinyl alcohol, moisturizing agent and ultrapure water and stir;
[0017] b. Add the engineered probiotics into the system obtained in step a, stir evenly, and obtain a probiotic antibacterial gel dressing.
[0018] Furthermore, in step a, the reaction temperature is 35-45° C. and the reaction time is 2-3 h.
[0019] Furthermore, in step b, the reaction temperature is 25-35° C. and the reaction time is 1-2 h.
[0020] The advantages and positive effects of the probiotic antibacterial gel dressing and the preparation method thereof described in the present invention are:
[0021] 1. The present invention uses Escherichia coli Nissle 1917 (Escherichia coli Nissle1917, EcN) and Poria cocos extract as active substances, avoiding the problem of drug resistance caused by the existing technology of using antibiotics; at the same time, Escherichia coli effectively inhibits the growth of pathogenic bacteria by releasing antimicrobial peptides, iron ion carriers, enzymes and other substances, and can also inhibit the colonization of pathogenic bacteria (such as enterohemorrhagic Escherichia coli) and the production of toxins (such as Shiga toxin-producing Escherichia coli); triterpenoid compounds and polysaccharides in Poria cocos extract as natural active substances have multiple physiological activities, such as improving the body's immunity, antibacterial, hypoglycemic, anti-tumor and the like.
[0022] 2. The Escherichia coli used in the probiotic antibacterial gel dressing prepared by the present invention can regulate inflammation at the wound. It regulates inflammation by affecting the expression of miRNA in the tissue and can promote wound recovery.
[0023] 3. The probiotic antibacterial gel dressing prepared by the present invention has good film-forming property. After being applied on the skin or the surface of the catheter, it will spontaneously form a thin film with good film-forming property, which can isolate the wound from the external environment, play a good moisturizing effect, and at the same time improve the colonization rate and survival rate of probiotics in the wound.
[0024] 4. The probiotic antibacterial gel dressing prepared by the present invention forms a network structure with excellent dispersibility during the emulsification and mixing process, has good toughness, is easy to be completely removed from the skin or the catheter surface, is easy to use, and has high patient compliance.
[0025] The technical solution of the present invention is further described in detail below through embodiments. DETAILED DESCRIPTION
[0026] The technical solution of the present invention is further illustrated by the following examples.
[0027] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.
[0028] The preparation steps of engineered probiotics are as follows:
[0029] S1. Centrifuge the E. coli cultured overnight in LB liquid medium and resuspend it in PBS;
[0030] S2, repeat step S1 three times, and resuspend the E. coli in 10 mM Tirs-HCl at pH 8.5 for the third time;
[0031] S3. Add 50 mg / mL dopamine hydrochloride to the E. coli solution and stir for 30 min;
[0032] S4, adding Poria cocos extract to the system of step S3, and stirring for 30 minutes;
[0033] S5. Centrifuge the system in step S4 and resuspend it in ice PBS to obtain engineered probiotics.
[0034] Example 1
[0035] In the above step S4, 50 mg / mL Poria cocos extract was added (the remaining steps were the same as the above steps) to construct the engineered probiotics.
[0036] Example 2
[0037] In the above step S4, 100 mg / mL of Poria cocos extract was added (the remaining steps were the same as the above steps) to construct the engineered probiotics.
[0038] Example 3
[0039] In the above step S4, 200 mg / mL of Poria cocos extract was added (the remaining steps were the same as the above steps) to construct the engineered probiotics.
[0040] The amount of Poria cocos extract added in Example 1, Example 2 and Example 3 is shown in Table 1.
[0041] Table 1
[0042] sample Example 1 Example 2 Example 3 Poria cocos extract amount 50mg / mL 100mg / mL 200mg / mL
[0043] Activity detection of engineered probiotics:
[0044] An equal amount of engineered probiotics was cultured on LB solid medium at 37°C for 12 h, and the number of bacterial colonies in the solid medium was recorded. This number was the activity of the engineered probiotics. The results are shown in Table 2.
[0045] Table 2
[0046] sample Example 1 Example 2 Example 3 Probiotic activity 48 52 43
[0047] It can be seen from Table 2 that as the amount of Poria extract loaded with probiotics increases, the activity of the engineered probiotics first increases and then decreases. Therefore, the loading amount of Poria extract of the engineered bacteria in the subsequent preparation of the gel dressing is 100 mg / mL.
[0048] The preparation steps of a probiotic antibacterial gel dressing are as follows:
[0049] a. Mix 5%-30% polyvinyl alcohol, 5%-20% moisturizer and the balance ultrapure water according to mass percentage, and stir;
[0050] b. Add 0.5%-5% of the engineered probiotics into the system obtained in step a according to the mass percentage, and stir to obtain the probiotic antibacterial gel dressing.
[0051] Example 4
[0052] 10% polyvinyl alcohol, 10% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0053] Example 5
[0054] 10% polyvinyl alcohol, 12% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0055] Example 6
[0056] 10% polyvinyl alcohol, 15% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0057] Example 7
[0058] 15% polyvinyl alcohol, 10% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0059] Example 8
[0060] 15% polyvinyl alcohol, 12% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0061] Example 9
[0062] 15% polyvinyl alcohol, 15% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0063] Example 10
[0064] 20% polyvinyl alcohol, 10% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0065] Embodiment 11
[0066] 20% polyvinyl alcohol, 12% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0067] Example 12
[0068] 20% polyvinyl alcohol, 15% moisturizer and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0069] The components of the probiotic antibacterial gel dressing are shown in Table 3.
[0070] Table 3 (Unit mass percentage)
[0071] sample Polyvinyl alcohol amount Moisturizing dosage water Example 4 10 10 margin Example 5 10 12 margin Example 6 10 15 margin Example 7 15 10 margin Example 8 15 12 margin Example 9 15 15 margin Example 10 20 10 margin Embodiment 11 20 12 margin Example 12 20 15 margin
[0072] Antibacterial gel dressing hardness test:
[0073] The effect of different polyvinyl alcohol contents on gel hardness was detected, and the hardness of gel dressings with different polyvinyl alcohol contents was recorded. The results are shown in Table 4.
[0074] Table 4
[0075] sample Example 4 Example 7 Example 10 Gel hardness Moderate Moderate Moderate
[0076] It can be seen from Table 4 that the antibacterial gel has moderate hardness and is suitable for application.
[0077] Antibacterial gel dressing moisturizing effect test:
[0078] The moisture content of the skin surface was tested. The antibacterial gel dressing was applied to the skin for 30 minutes. The skin moisture content was tested again after 30 minutes. The skin moisture content before and after the use of the gel dressing was recorded. The results are shown in Table 5.
[0079] Table 5
[0080] sample Example 7 Example 8 Example 9 Skin moisture content before use 17.5％ 18.1％ 17.3％ Skin moisture content after use 18.1％ 18.8％ 18.1％ Increased skin moisture content 0.6％ 0.7％ 0.8％
[0081] As shown in Table 5, the antibacterial gel can significantly increase the water content of the skin.
[0082] Example 13
[0083] 15% polyvinyl alcohol, 12% moisturizer, 2% engineered probiotics, and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0084] Embodiment 14
[0085] 15% polyvinyl alcohol, 12% moisturizer, 5% engineered probiotics, and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0086] Embodiment 15
[0087] 15% polyvinyl alcohol, 12% moisturizer, 10% engineered probiotics, and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0088] Comparative Example 1
[0089] 15% polyvinyl alcohol, 12% moisturizer, 0% engineered probiotics, and the balance ultrapure water (the remaining steps are the same as the above steps) to obtain a probiotic antibacterial gel dressing.
[0090] The components of the probiotic antibacterial gel dressing containing different amounts of probiotics are shown in Table 6.
[0091] Table 6 (Unit mass percentage)
[0092] sample Example 13 Embodiment 14 Embodiment 15 Comparative Example 1 Probiotics 2 5 10 0 Polyvinyl alcohol 15 15 15 15 Moisturizer 12 12 12 12 water margin margin margin margin
[0093] Antibacterial gel dressing antibacterial test:
[0094] Take 1mL for about 10 6 A CFU / mL concentration of Staphylococcus aureus suspension was mixed with 1 g of probiotic antibacterial gel dressing and cultured at 37°C for 12 h. The cultured solution was diluted 100 times, and 1 mL of the solution was inoculated into LB broth solid culture medium. The culture was incubated at 37°C for 12 h, and the number of bacteria on the solid culture medium was recorded. The results are shown in Table 7.
[0095] Table 7 (unit)
[0096] sample Example 13 Embodiment 14 Embodiment 15 Comparative Example 1 Colony count 29 11 6 198
[0097] Table 7 is the antibacterial test results of the antibacterial gel dressing of the present invention. It can be seen from Table 7 that the antibacterial effect of Example 15 is significantly better than that of Control Example 1. The probiotic antibacterial gel dressing can effectively inhibit the growth of Staphylococcus aureus. At the same time, it can be seen that with the increase in the amount of probiotics added, the antibacterial gel dressing has a better effect in inhibiting the growth of Staphylococcus aureus.
[0098] Film-forming property test of antibacterial gel dressing:
[0099] The probiotic antibacterial gel dressing has good film-forming property and the film is easy to be completely removed.
[0100] Antibacterial gel dressing probiotic activity test:
[0101] The activity of probiotics was tested on the 1st, 7th, 15th and 30th days of the prepared antibacterial gel dressing. The test method was as follows: 1±0.1 g of the dressing was soaked in 50 mL of pH=7 PBS solution for 2 h, homogenized at 2000 rpm for 5 min, dialyzed with pH=7 PBS solution for 3 d (dialysis bag was 10000 Da), replaced every 12 h, and then the contents were centrifuged, 1 mL of the supernatant was diluted 10 times, 1 mL of the dilution was inoculated into LB broth solid culture medium, and cultured at 37°C for 12 h, and the number of bacteria on the solid culture medium was recorded. The results are shown in Table 8.
[0102] Table 8 (Unit: LogCFU / mL)
[0103] project Day 1 Day 7 Day 15 Day 30 Example 13 4.28 4.11 4.05 3.98 Embodiment 14 10.7 10.27 10.12 9.95 Embodiment 15 32.1 30.82 30.37 29.85
[0104] Table 8 is the test result of probiotic activity of antibacterial gel dressing. It can be seen from Table 8 that the activity of probiotics in the antibacterial gel dressing decreases with the extension of time. The decrease in activity is not obvious, but it can still maintain a high activity.
[0105] Therefore, the present invention adopts the above-mentioned probiotic antibacterial gel dressing and its preparation method, which has the advantages of long-lasting antibacterial ability, low toxicity, no drug resistance, and no side effects. It can be applied to wound dressings, catheter infections, gastrointestinal infections and other fields, and can solve the problems of long-term use of antibacterial gel dressings in the prior art causing drug resistance, cumulative toxicity, and low antibacterial persistence.
[0106] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.
Claims
1. A probiotic antibacterial gel dressing, characterized in that: The invention comprises the following components by weight: 5% to 25% polyvinyl alcohol, 0.5% to 5% engineered probiotics, 5% to 20% moisturizer, and the balance is ultrapure water; wherein the engineered probiotics include Poria cocos extract, dopamine and Escherichia coli, the effective components of Poria cocos extract include polysaccharides, triterpenes, fatty acids, sterols, enzymes, and the Escherichia coli is Escherichia coli Nissle 1917; The preparation method of the engineered probiotics is as follows: S1. Centrifuge the E. coli cultured overnight in LB liquid medium and resuspend it in PBS; S2, repeat step S1 three times, and resuspend the E. coli in 10 mM Tirs-HCl at pH 8.5 for the third time; S3. Add 50 mg / mL dopamine hydrochloride to the E. coli solution and stir for 30 min; S4, adding 100 mg / mL Poria extract to the solution of step S3, and stirring for 30 min; S5. Centrifuge the system in step S4 and resuspend it in ice PBS to obtain engineered probiotics.
2. The probiotic antibacterial gel dressing according to claim 1, characterized in that: The invention comprises the following components by weight: polyvinyl alcohol 10% to 20%, engineered probiotics 2% to 5%, moisturizer 12% to 15%, and the balance is ultrapure water.
3. The probiotic antibacterial gel dressing according to claim 1, characterized in that: The moisturizing agent is one or more of 1,3-propylene glycol, glycerol, 1,3-butylene glycol, sorbitol and polyethylene glycol.
4. A method for preparing a probiotic antibacterial gel dressing according to any one of claims 1 to 3, characterized in that: The following steps are involved: a. Mix polyvinyl alcohol, moisturizing agent and ultrapure water and stir; b. Add the engineered probiotics into the system obtained in step a, stir evenly, and obtain a probiotic antibacterial gel dressing.
5. The preparation method according to claim 4, characterized in that: In step a, the reaction temperature is 35-45° C. and the reaction time is 2-3 h.
6. The preparation method according to claim 4, characterized in that: In step b, the reaction temperature is 25-35° C. and the reaction time is 1-2 h.