CN113476482A - Probiotic external preparation - Google Patents
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- CN113476482A CN113476482A CN202110923862.1A CN202110923862A CN113476482A CN 113476482 A CN113476482 A CN 113476482A CN 202110923862 A CN202110923862 A CN 202110923862A CN 113476482 A CN113476482 A CN 113476482A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 41
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- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical group CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 16
- 239000000661 sodium alginate Substances 0.000 claims description 16
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
- A61K35/64—Insects, e.g. bees, wasps or fleas
- A61K35/644—Beeswax; Propolis; Royal jelly; Honey
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/02—Local antiseptics
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Abstract
The invention provides a probiotic external preparation, which comprises 5mL of ointment and the concentration of the ointment is 107‑1010CFU/mL probiotics, wherein the ointment consists of 20% by volume of ointment matrix and 5% -35% by volume of honey. The invention mixes the probiotic preparation and the honey for the first time, and the synergistic effect of the probiotic preparation and the honey can better play the role of inhibition and antagonismAnd (3) an anti-pathogenic microorganism effect.
Description
Technical Field
The invention belongs to the technical field of medical use, and particularly relates to a probiotic external preparation.
Background
Probiotics are the focus of scientific research in recent years. The probiotics as the component of the normal flora has the functions of improving the immunity of human bodies, regulating the flora balance, reducing blood cholesterol and the like, and plays a very important role in maintaining the health of the human bodies. Wherein, the probiotics are fixed on the body surface and the surfaces of intestinal tracts, oral cavities, vaginas and the like, and play an important role in preventing the colonization and infection of pathogenic microorganisms through immunoregulation, secretion of antagonistic substances and colonization of antagonistic pathogenic bacteria. Therefore, the probiotic preparation has potential good effects and potential for further research and development into external preparations for infecting the wound surface in the aspects of preventing and treating skin wound surface infection, antagonizing pathogenic microorganism infection and promoting wound surface healing.
Honey, as a commonly used Chinese medicine, has sweet taste and mild nature and enters lung, spleen and large intestine meridians. Since ancient times, honey is used as an external medicine for aphthous erosion, boil from hot water, red eyes with swelling and pain, sores, carbuncles, furuncles and the like; modern medical research shows that the honey has the effects of antagonizing and inhibiting pathogenic microorganisms and promoting wound healing, and can be used for treating wounds, burn wounds, skin ulcers and the like.
Many researches prove that the honey has various biological activities such as antibiosis, antioxidation, anti-inflammation and the like. However, few studies have explored the use of probiotics in the treatment of refractory wounds for antibacterial and anti-inflammatory ointments.
Disclosure of Invention
In view of the above-mentioned disadvantages in the prior art, the present invention aims to provide a probiotic external preparation, which can be applied to the treatment of refractory wounds.
In order to achieve the purpose, the invention provides the following technical scheme: a probiotic external preparation comprises 5mL of ointment and the concentration of the ointment is 107-1010CFU/mL probiotics, wherein the ointment consists of 20% by volume of ointment matrix and 5% -35% by volume of honey.
Further, the concentration of the probiotics is 109CFU/mL, the ointment is composed of 20% by volume of ointment matrix and 5% by volume of honey.
Further, the probiotics are lactobacillus plantarum or lactobacillus rhamnosus.
Further, the ointment base is sodium alginate.
The probiotic external preparation is prepared by adopting the following method:
(1) firstly, heating the ointment matrix in a water bath at 80 ℃ by using sterile ultrapure water until the ointment matrix is dissolved to obtain jelly;
(2) adding the above Mel into the jelly, and making into ointment;
(3) inoculating the probiotics into MRS broth, and standing and culturing at 37 ℃ for 16 h; adding the cultured probiotics into the ointment of the step (2), and culturing at 37 ℃ for 24 h.
Further, the ointment base in the step (1) is sterilized by gamma irradiation of 25kGy before use.
Further, before the honey in the step (2) is used, the honey is subjected to sterilization treatment by gamma ray irradiation of 25 kGy.
Compared with the prior art, the invention has the following beneficial effects:
(1) the probiotic preparation and the honey are mixed for the first time, and the synergistic effect of the probiotic preparation and the honey can better play the effect of inhibiting and antagonizing pathogenic microorganisms, and is superior to the effect of conventional antibiotics.
(2) The main component of the invention is biological, which can be absorbed and decomposed by organism, without toxic and side reaction of inorganic compound (disinfectant) for long-term use.
(3) The probiotic external preparation provided by the invention can enlarge the use effect of probiotics, provides scientific basis for further research and development of safe and effective probiotic preparations for wounds infected with wound surfaces which are difficult to heal, and has important social and economic values.
Drawings
FIG. 1 is a graph of the effect of different honey concentrations on the growth of probiotics as well as Staphylococcus aureus;
FIG. 2 shows the inhibitory effect of different probiotic concentrations on Staphylococcus aureus.
Detailed Description
The process of the present invention will be described in detail with reference to specific examples.
First, preparation of probiotic external preparation
Example 1
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration7CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 5% by volume honey.
Example 2
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration8CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 5% by volume honey.
Example 3
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration9CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 5% by volume honey.
Example 4
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration10CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 5% by volume honey.
Example 5
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration7CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 20% by volume honey.
Example 6
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration8CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 20% by volume honey.
Example 7
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration9CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 20% by volume honey.
Example 8
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration10CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 20% by volume honey.
Example 9
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration7CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 35% by volume honey.
Example 10
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration8CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 35% by volume honey.
Example 11
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration9CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 35% by volume honey.
Example 12
A probiotic bacteria external preparation comprises 5mL of ointment and 10% concentration10CFU/mL lactobacillus plantarum, the ointment consisting of 20% by volume sodium alginate and 35% by volume honey.
The probiotic bacteria external preparation of the above examples 1 to 12 was prepared by the following method:
(1) firstly, heating the ointment matrix in a water bath at 80 ℃ by using sterile ultrapure water until the ointment matrix is dissolved to obtain jelly;
(2) adding the above Mel into the jelly, and making into ointment;
(3) inoculating the probiotics into MRS broth, and standing and culturing at 37 ℃ for 16 h; adding the cultured probiotics into the ointment of the step (2), and culturing at 37 ℃ for 24 h.
The ointment matrix in the invention is sodium alginate, and the sodium alginate and the honey are sterilized by gamma-ray irradiation of 25kGy before use. In addition, the honey in the invention is prepared into solution by adopting water.
The probiotic bacteria in the present invention may also be lactobacillus rhamnosus.
The ointment base in the present invention may also be polyethylene glycols, vaseline, lanolin or liquid paraffin.
Second, determination of honey concentration
2.1) Effect of ointments on probiotic growth
Inoculating the probiotics into MRS broth, and performing static culture at 37 ℃ for 16 h. Adding cultured probiotic into ointment containing Mel at different ratio, culturing at 37 deg.C for 24h, diluting at multiple ratio, coating MRS agar plate, making two parallel dilutions, and determining the influence of ointment on probiotic growth. Selecting an ointment formula with the minimum influence on the growth of the probiotics to prepare the probiotic honey preparation.
2.2) Effect of ointments on the growth of Staphylococcus aureus
Staphylococcus aureus was inoculated into LB broth, incubated overnight at 37 ℃ on a shaker. After the bacterial suspension is diluted to 10CFU/mL, 100ul of bacterial suspension is added into a 96-well plate, and the plate is cultured overnight at 37 ℃, so that staphylococcus aureus forms a biofilm at the bottom of the 96-well plate. Then, the bacterial liquid in the holes is sucked out, 100ul of ointments with different honey concentrations are respectively added into the holes, incubation is carried out at 37 ℃, after 24 hours, the culture is taken, and the influence of the ointments on the growth of staphylococcus aureus is detected by a dilution plate method.
The influence of the ointment on the growth of probiotics and staphylococcus aureus is respectively measured, and an ointment formula which has small influence on the growth of the probiotics and good growth inhibition effect on the staphylococcus aureus is selected. The results are shown in FIG. 1. Figure 1 shows the effect of different honey concentrations on the growth of probiotics as well as staphylococcus aureus, where the measurements are expressed as mean ± standard error (mean ± SEM), P <0.001 and P <0.0001 compared to the initial concentration group of bacteria.
As can be seen from fig. 1, in the experiment for determining the ointment formula, the result shows that the concentration of honey in the ointment has little influence on the growth of probiotics, but has larger influence on the growth of staphylococcus aureus, and the inhibitory effect on staphylococcus aureus is stronger with the increase of the concentration of honey. Comprehensively considering, an ointment formula with 5% honey concentration is selected.
Thirdly, the probiotic external preparation of the invention has the inhibiting effect on staphylococcus aureus
Selecting ointment with minimal influence on growth of probiotic bacteria, adding probiotic bacteria with different concentrations, and respectively preparing into ointment with bacteria content of 107、108、109、1010CFU/mL of probiotic honey external preparation is used for measuring the inhibition effect on staphylococcus aureus in vitro.
Taking 96-well plate, adding Staphylococcus aureus 105The nutrient broth was inoculated at a concentration of CFU/mL, added to a 96-well plate at 100ul per well and incubated overnight at 37 ℃ to allow Staphylococcus aureus to form a biofilm on the bottom of the 96-well plate. Then absorbing bacterial liquid in the holes, respectively adding 100ul of honey preparations containing probiotics with different concentrations into the holes, simultaneously setting 3 positive controls which are respectively blank ointment matrix, blank ointment matrix and honey, blank ointment matrix and staphylococcus aureus antibiotic ampicillin sodium, culturing for 24h and 48h at 37 ℃, taking the culture, coating a Baird-Parker plate after dilution, making two parallels for each dilution degree, and detecting the growth and survival state of the staphylococcus aureus.
By detecting the growth and survival state of staphylococcus aureus, the result of selecting the honey preparation with the probiotic concentration having the best inhibitory effect on staphylococcus aureus is shown in fig. 2. Fig. 2 is a graph of inhibition of staphylococcus aureus by probiotic honey formulations, wherein the dosing data are expressed as mean ± standard error (mean ± SEM), # P <0.05, # P <0.01, # P <0.001, # P <0.05, and # P <0.05 compared to the initial concentration group of bacteria.
By detecting the growth and survival state of staphylococcus aureus and selecting the probiotic honey preparation with the best inhibiting effect on staphylococcus aureus, the result shows that the inhibiting effect of the probiotic honey preparation on staphylococcus aureus is gradually enhanced along with the increase of the concentration of the probiotic, and compared with the probiotic with low concentration, the probiotic honey preparation is 10 percent9The inhibiting effect of the CFU/mL probiotics on staphylococcus aureus is more obvious, so that the concentration of the selected probiotics is 109CFU/mL honey external preparation.
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
Claims (7)
1. The probiotic external preparation is characterized by comprising 5mL of ointment and 10 concentration7-1010CFU/mL probiotics, wherein the ointment consists of 20% by volume of ointment matrix and 5% -35% by volume of honey.
2. The probiotic external preparation according to claim 1, characterized in that the concentration of the probiotic is 109CFU/mL, the ointment is composed of 20% by volume of ointment matrix and 5% by volume of honey.
3. The probiotic external preparation according to claim 1, characterized in that the probiotic is Lactobacillus plantarum or Lactobacillus rhamnosus.
4. The probiotic external preparation according to claim 1, characterized in that the ointment base is sodium alginate, polyethylene glycols, vaseline, lanolin or liquid paraffin.
5. The method for preparing the probiotic external preparation according to any one of claims 1 to 4, characterized by comprising the steps of:
(1) firstly, heating the ointment matrix in a water bath at 80 ℃ by using sterile ultrapure water until the ointment matrix is dissolved to obtain jelly;
(2) adding the above Mel into the jelly, and making into ointment;
(3) inoculating the probiotics into MRS broth, and standing and culturing at 37 ℃ for 16 h; adding the cultured probiotics into the ointment of the step (2), and culturing at 37 ℃ for 24 h.
6. The method for preparing the probiotic external preparation according to claim 5, wherein the ointment base in the step (1) is sterilized by gamma irradiation of 25kGy before use.
7. The method for preparing the probiotic external preparation according to claim 5, wherein the honey in the step (2) is sterilized by gamma irradiation of 25kGy before use.
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| US20190343901A1 (en) * | 2016-12-30 | 2019-11-14 | Northwestern University | Vehicles for Applying Bacteria to Skin, Scalp, and Hair |
| CN110964665A (en) * | 2019-12-20 | 2020-04-07 | 哈尔滨美华生物技术股份有限公司 | Probiotic composition for post-gastrectomy treatment and preparation method thereof |
| US20210386803A1 (en) * | 2016-12-30 | 2021-12-16 | Ann And Robert H. Lurie Children's Hospital Of Chicago | Vehicles for applying bacteria to skin, scalp, and hair |
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