CN115252757A - Marine organism physical barrier wound protection film and preparation method thereof - Google Patents
Marine organism physical barrier wound protection film and preparation method thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/02—Peptides of undefined number of amino acids; Derivatives thereof
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- 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
- A61K35/02—Medicinal preparations containing materials or reaction products thereof with undetermined constitution from inanimate materials
- A61K35/10—Peat; Amber; Turf; Humus
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- 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
- A61K35/60—Fish, e.g. seahorses; Fish eggs
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
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- 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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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7015—Drug-containing film-forming compositions, e.g. spray-on
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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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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention relates to a marine biophysical barrier wound protection film and a preparation method thereof, belonging to the field of medical materials. The invention relates to a marine organism physical barrier wound-protecting film which comprises mussel extract, grass green salicornia extract, ethanol, thermophilic thermus bacteria fermentation product, caprylic acid/capric acid triglyceride, VP/VA copolymer, laminaria intermedia extract, polyethylene glycol-8, undaria pinnatifida extract, macadamia mesh fermentation product, laminaria saccharina extract, sturgeon caviar extract, phenoxyethanol, potassium sorbate, glycerol, tocopherol and deionized water, and the production process steps and parameters are optimized, so that the wound-protecting film containing marine organism components and used for repairing the skin physical barrier is obtained. Wherein, the mussel extract, the laminaria cuneata extract and the sturgeon caviar extract have synergistic effect, and can repair skin barrier and promote wound healing. The physical barrier wound-protecting film of marine organisms has good biocompatibility, and can promote skin repair and fade scars.
Description
Technical Field
The invention belongs to the field of medical materials, and particularly relates to a marine organism physical barrier wound protection film and a preparation method thereof.
Background
The skin is the first line of defense of the human body, has important barrier function, is mainly formed by emulsifying sebum secreted by sebaceous glands, lipid produced by disintegration of cells of stratum corneum and sweat secreted by sweat glands, and a water-fat film and a brick wall structure jointly form a physical barrier of the skin, so that the loss of water and electrolyte in the body is prevented, the invasion of harmful substances can be prevented, and the maintenance of homeostasis in the body is facilitated. The homeostasis of the skin barrier is a complex network regulation process, including various pathophysiological processes such as the growth, differentiation, immune response and inflammation of skin cells. In daily life, mechanical, physical and chemical factors all cause damage to the skin barrier. The wound protecting film is a biological film capable of replacing gauze accessories, can protect the wound surface from infection, and has better effect when being used for patients with large-area burn. The wound protecting film obtained by the existing preparation method has great differences in biocompatibility, physiological effect and the like, and needs to be continuously adjusted and optimized.
The invention discloses an antibacterial wound protection film disclosed in patent publication No. CN 105641480A, which consists of cattail pollen, pseudo-ginseng, folium artemisiae argyi, sophora flower, lalang grass rhizome, chitosan, glycerol and distilled water. However, the biocompatibility, the effective repair of the physical barrier of the skin and the scar lightening are poor.
Aiming at solving the problems that serious infection is easy to cause and wounds are difficult to heal due to skin barrier defect, the invention prepares the marine biophysical barrier wound-protecting film loaded with mussel mucin (mussel extract). The ocean contains rich biological resources and is a natural resource treasury. The marine organism raw material has unique functions and various purposes. Marine proteins (mussel mucin) and grass green Salicornia herbacea extract, thermus thermophilus fermentation product, laminaria ochracea extract, undaria pinnatifida extract, laminaria saccharina extract, sea sludge extract, and sturgeon caviar extract have good biological functionality and biological safety, and have been widely used as cosmetic raw materials. The invention screens the raw material components of the product of the marine organism physical barrier wound-protecting film, optimizes the production process steps and parameters, development has resulted in a new wound-care film containing marine biological components to repair the physical barrier of the skin. The wound protection film has the advantages of convenience in use, wound surface protection, acceleration of wound healing and the like, and can effectively repair physical barriers of the skin.
Disclosure of Invention
The invention aims to provide a marine organism physical barrier wound protection film and a preparation method thereof, belonging to the field of medical materials. The invention relates to a novel wound protection film containing marine biological components for repairing skin physical barriers, which is developed by screening raw material components of the wound protection film of the marine biological physical barriers, selecting mussel extract, grass green salicornia extract, ethanol, thermophilic thermus thermophilus fermentation product, caprylic/capric triglyceride, VP/VA copolymer, laminaria cuneata extract, polyethylene glycol-8, undaria pinnatifida extract, macadamia schreberi mud fermentation product, laminaria saccharina extract, sturgeon caviar extract, phenoxyethanol, potassium sorbate, glycerol, tocopherol and deionized water, and optimizing the production process steps and parameters. Wherein, the mussel extract, the laminaria cuneata extract and the sturgeon caviar extract have synergistic effect, and can repair skin barrier and promote wound healing. The marine organism physical barrier wound protection film has the advantages of good biocompatibility, capability of promoting skin repair, scar fading, good safety and the like, can be sprayed on a wound surface or smeared on the wound surface, and is convenient to use.
The purpose of the invention can be realized by the following technical scheme:
a marine biophysical barrier wound-protecting film comprises 0.01-0.2 part by weight of mussel extract, 0.01-0.2 part by weight of turquoise salicornia extract, 30-60 parts by weight of ethanol, 0.01-5 parts by weight of Thermus thermophilus fermentation product, 10-30 parts by weight of caprylic/capric triglyceride, 1-10 parts by weight of VP/VA copolymer, 0.05-1 part by weight of Laminaria angustifolia extract, 10-30 parts by weight of polyethylene glycol-8, 0.01-0.2 part by weight of Undaria pinnatifida extract, 0.01-2 parts by weight of Hawaii mud fermentation product, 0.01-2 parts by weight of kelp extract, 0.01-1 part by weight of sturgeon roe extract, 0.01-0.05 part by weight of phenoxyethanol, 0.01-0.02 part by weight of potassium sorbate, 1-10 parts by weight of glycerol, 0.01-0.02 part by weight of tocopherol and 1-10 parts by weight of deionized water.
As a preferred technical scheme of the invention, the method for extracting the mussel extract from the wound protecting film of the marine biophysical barrier comprises the following steps:
(1) Putting 1 weight part of crushed mussel foot silk gland and 10 weight parts of deionized water into an extraction tank for extraction to obtain an extracting solution;
(2) Adding 0.01 to 0.05 times weight of protease into the extracting solution, stirring uniformly, and carrying out enzymolysis for 1 to 2 hours in a constant-temperature water bath kettle at the temperature of between 40 and 50 ℃ to obtain an enzymolysis product;
(3) Filtering the enzymolysis product with 200 mesh filter screen, and concentrating the filtrate to obtain mussel extract.
As a preferred technical scheme of the invention, the preparation method of the laminaria cuneata extract comprises the following steps:
1) Soaking the laminaria cuneata in warm water at 45 ℃ for 3-5 h, and then cleaning;
2) Taking the cleaned laminaria cuneata in the step 1), crushing by using a crusher, adding pectinase, protease and cellulase which are 0.9 times the weight of the laminaria cuneata and have the weight ratio of 1;
3) Taking the enzymolysis liquid in the step 2) to inactivate enzyme for 5-8 s at 100-120 ℃ to obtain an inactivated enzyme liquid;
4) Centrifuging the enzyme-inactivating liquid obtained in the step 3) at 6000-8000 rpm for 8-10 min to obtain supernatant;
5) Concentrating the supernatant in the step 4), and performing spray drying to obtain the laminaria cuneata extract.
As a preferred technical scheme of the invention, the preparation method of the sturgeon caviar extract comprises the following steps:
A. crushing sturgeon caviar, and then adding 8-10 times of treatment fluid by weight to obtain treated sturgeon caviar;
B. cleaning the sturgeon caviar treated in the step A, adding 1-2 times of deionized water by weight, adding 0.01 times of enzymolysis liquid by weight, and carrying out enzymolysis at 40-50 ℃ for 2-3 h to obtain an enzymolysis product;
C. b, taking the enzymolysis product in the step B, and inactivating enzyme for 5-8 s at 100-120 ℃ to obtain an enzyme inactivation solution;
D. centrifuging the enzyme-inactivating liquid obtained in the step C at 6000-8000 rpm for 8-10 min to obtain supernatant;
E. and D, concentrating the supernatant in the step D, and freeze-drying to obtain the sturgeon caviar extract.
As a preferable technical scheme of the invention, the treatment solution in the step A in the preparation method of the sturgeon caviar extract comprises 3-5 parts by weight of sodium chloride solution, 0.5-1 part by weight of hydrogen peroxide solution, 0.1-0.5 part by weight of calcium chloride solution, 0.1-0.5 part by weight of glacial acetic acid solution and 60-80 parts by weight of deionized water.
As a preferable technical scheme of the present invention, the enzymolysis solution of step B in the preparation method of the sturgeon caviar extract comprises 1 part by weight of papain, 1 part by weight of alkaline protease, 1 part by weight of lecithinase and 1 part by weight of neutral protease.
In a preferred embodiment of the present invention, the Hawaii sea mud fermentation product in the physical barrier wound-protecting film of marine organisms comprises 25 parts by weight of sea mud extract, 2 parts by weight of Leuconostoc/radish root fermentation product filtrate, and 73 parts by weight of glycerin.
The preparation method of the marine organism physical barrier wound protection film comprises the following steps:
s1, adding ethanol into a water phase pot, then slowly adding the VP/VA copolymer into the water phase pot, and stirring until the VP/VA copolymer is uniformly dissolved.
S2, sequentially putting the mussel extract, the carex glauca extract, polyethylene glycol-8, the thermophilic thermus bacterium fermentation product, the laminaria cuneata extract, caprylic/capric triglyceride, the undaria pinnatifida extract, the macadamia javanica mud fermentation product, the laminaria saccharina extract, the sturgeon caviar extract, phenoxyethanol, potassium sorbate, glycerol, tocopherol and deionized water into a water phase pot, and stirring and dispersing uniformly.
S3, after stirring uniformly, discharging after checking the pH value to be 5-6.5.
As a preferred technical scheme of the invention, the temperature of the water phase pot in the preparation method of the marine organism physical barrier wound-protecting film is controlled to be about 20-25 ℃.
The invention has the beneficial effects that:
(1) The invention simulates a skin physical barrier to form a protective film which is air-permeable, bacterium-impermeable, soft, elastic, good in biocompatibility and capable of promoting wound healing, and additionally loads marine organism repairing and moisturizing components of a carex incarnata extract, a thermophilic thermus strain fermentation product, a laminaria cuneata extract, an undaria pinnatifida extract, a laminaria saccharina extract, a sea sludge extract, a sturgeon caviar extract and the like, and combines a mussel extract molecular structure containing a high content of dopa groups, so that oxidative crosslinking can be performed in the air to realize bonding. The macroscopic adhesion effect is microscopically embodied in promoting the adherence, crawling and growth of cells, protecting the skin from the invasion of microorganisms, being not easy to be stimulated by the outside and playing a role of physical barrier.
(2) The mussel extract is a protein (mussel mucin) extracted and purified from the byssus gland of marine organism mussel, has strong adhesive effect, and is a biological glue. Mussel mucin is combined with cells through electrostatic interaction, so that adherence and creeping substitution of epidermal cells are promoted, and healing of a wound surface or an ulcer surface is accelerated. Several amino acids constituting mussel mucin contain hydrophobic groups, and in the presence of physiological solution, part of the hydrophobic groups of mussel mucin are exposed and can be combined with the lipid bilayer of cell membrane through hydrophobic interaction. Mussel mucin has potential repairing function on the wound surface of skin, mucous membrane tissue, nerve, bone and blood vessel membrane. Mussel mucin has certain advantages over other medical repair materials and drugs in its biological origin and the nature of the protein. Recent studies have shown that mussel mucin can also be used for surgical incisions, burns, scalds, persistent ulcers and skin diseases, and has analgesic, bacteriostatic and healing promoting effects.
(3) The foreign biophysical barrier wound protection film prepared by the invention is a fine fiber filament like a silk, a fine transparent film is formed on the surface of the skin, and the skin is attached to the skin as if the second layer of the skin is common. Wherein, the mussel extract, the laminaria cuneata extract and the sturgeon caviar extract have synergistic effect, and can repair skin barrier and promote wound healing. The marine biophysical barrier wound protection film has the advantages of good biocompatibility, promotion of skin repair, scar fading, good safety and the like, can be sprayed on a wound surface or smeared on the wound surface, and is convenient to use.
Detailed Description
To further illustrate the technical means and effects of the present invention adopted to achieve the predetermined objects, the following detailed description of the embodiments, structures, features and effects according to the present invention will be provided in conjunction with the embodiments.
Example 1
In the specific embodiment, the weight ratios of the components of each phase are as follows:
0.2 parts by weight of mussel extract, 0.2 parts by weight of carex sibiricus extract, 30 parts by weight of ethanol, 5 parts by weight of thermus thermophilus fermentation product, 10 parts by weight of caprylic/capric triglyceride, 5 parts by weight of VP/VA copolymer, 1 part by weight of laminaria cuneata extract, 10 parts by weight of polyethylene glycol-8, 0.2 parts by weight of undaria pinnatifida extract, 2 parts by weight of macadamia sea mud fermentation product, 2 parts by weight of laminaria saccharina extract, 1 part by weight of sturgeon caviar extract, 0.01 part by weight of phenoxyethanol, 0.01 part by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 part by weight of tocopherol, and 10 parts by weight of deionized water.
The Hawaii sea mud fermentation product in the marine organism physical barrier wound protection film comprises 25 parts by weight of sea mud extract, 2 parts by weight of Leuconostoc/radish root fermentation product filtrate and 73 parts by weight of glycerol.
The preparation method of the marine organism physical barrier wound protection film comprises the following steps:
s1, adding 30 parts by weight of ethanol into an aqueous phase pot, slowly adding 5 parts by weight of VP/VA copolymer into the aqueous phase pot, and stirring until the VP/VA copolymer is uniformly dissolved;
s2, sequentially putting 0.2 part by weight of mussel extract, 0.2 part by weight of salicornia herbacea extract, 10 parts by weight of polyethylene glycol-8, 5 parts by weight of thermophilic thermus thermophilus fermentation product, 1 part by weight of laminaria cuneata extract, 10 parts by weight of caprylic/capric triglyceride, 0.2 part by weight of undaria pinnatifida extract, 2 parts by weight of macadamia sea mud fermentation product, 2 parts by weight of laminaria saccharina extract, 1 part by weight of sturgeon seed paste extract, 0.01 part by weight of phenoxyethanol, 0.01 part by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 part by weight of tocopherol and 10 parts by weight of deionized water into a water phase pot, and stirring and dispersing uniformly;
and S3, after uniformly stirring, checking the pH value to be 6, and then discharging.
During the preparation steps, the temperature of the water phase pot is controlled at 25 ℃.
The extraction method of the mussel extract comprises the following steps:
(1) Putting 1 weight part of crushed mussel foot silk glands and 10 weight parts of deionized water in an extraction tank for extraction to obtain an extracting solution;
(2) Adding protease 0.02 times the weight of the extract, stirring well, and performing enzymolysis in a water bath kettle at 40 deg.C for 2h to obtain enzymolysis product;
(3) Filtering the enzymolysis product with 200 mesh filter screen, and concentrating the filtrate to obtain mussel extract.
The extraction method of the laminaria cuneata extract comprises the following steps:
1) Soaking the laminaria cuneata in warm water at 45 ℃ for 5h, and then cleaning;
2) Crushing the cleaned laminaria cuneata in the step 1) by using a crusher, adding pectinase, protease and cellulase which are 0.9 times of the weight of the laminaria cuneata and have the weight ratio of 1;
3) Taking the enzymolysis liquid in the step 2) to inactivate enzyme for 5s at 100 ℃ to obtain an inactivated enzyme liquid;
4) Centrifuging the enzyme-inactivating liquid obtained in the step 3) at 6000rpm for 10min to obtain a supernatant;
5) Concentrating the supernatant in the step 4), and spray drying to obtain the laminaria cuneata extract.
The extraction method of the sturgeon caviar extract comprises the following steps:
A. crushing sturgeon caviar, and then adding 10 times of treatment liquid by weight to obtain treated sturgeon caviar;
B. cleaning the sturgeon caviar treated in the step A, adding 1 time of deionized water by weight, adding 0.01 time of enzymolysis liquid by weight, and carrying out enzymolysis for 2 hours at 40 ℃ to obtain an enzymolysis product;
C. b, taking the enzymolysis product in the step B, and inactivating the enzyme for 5s at 100 ℃ to obtain an enzyme inactivation solution;
D. c, taking the enzyme inactivation liquid obtained in the step C, and centrifuging at 6000rpm for 10min to obtain a supernatant;
E. and D, concentrating the supernatant in the step D, and then freezing and drying to obtain the sturgeon caviar extract.
In the preparation method of the sturgeon caviar extract, the treatment fluid in the step A comprises 3 parts by weight of sodium chloride, 1 part by weight of hydrogen peroxide, 0.5 part by weight of calcium chloride, 0.5 part by weight of glacial acetic acid and 60 parts by weight of deionized water.
The enzymolysis solution in the step B in the preparation method of the sturgeon caviar extract comprises 1 weight part of papain, 1 weight part of alkaline protease, 1 weight part of lecithinase and 1 weight part of neutral protease.
Comparative example 1
In the comparative example, the weight ratios of the components of each phase are respectively as follows:
0.2 parts by weight of a grass green salicornia extract, 30 parts by weight of ethanol, 5 parts by weight of a thermus thermophilus fermentation product, 10 parts by weight of caprylic/capric triglyceride, 5 parts by weight of a VP/VA copolymer, 1.2 parts by weight of a laminaria cuneata extract, 10 parts by weight of polyethylene glycol-8, 0.2 parts by weight of an undaria pinnatifida extract, 2 parts by weight of a macadamia mud fermentation product, 2 parts by weight of a laminaria saccharina extract, 1 part by weight of a sturgeon caviar extract, 0.01 parts by weight of phenoxyethanol, 0.01 parts by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 parts by weight of tocopherol, and 10 parts by weight of deionized water.
The Hawaii sea mud fermentation product in the marine organism physical barrier wound protection film comprises 25 parts by weight of sea mud extract, 2 parts by weight of Leuconostoc/radish root fermentation product filtrate and 73 parts by weight of glycerol.
The preparation method of the marine organism physical barrier wound protection film comprises the following steps:
s1, adding 30 parts by weight of ethanol into an aqueous phase pot, slowly adding 5 parts by weight of VP/VA copolymer into the aqueous phase pot, and stirring until the VP/VA copolymer is uniformly dissolved;
s2, sequentially putting 0.2 part by weight of green salicornia extract, 10 parts by weight of polyethylene glycol-8, 5 parts by weight of thermus thermophilus fermentation product, 1.2 parts by weight of laminaria cuneata extract, 10 parts by weight of caprylic/capric triglyceride, 0.2 part by weight of undaria pinnatifida extract, 2 parts by weight of macadamia fortunei mud fermentation product, 2 parts by weight of laminaria saccharina extract, 1 part by weight of sturgeon seed paste extract, 0.01 part by weight of phenoxyethanol, 0.01 part by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 part by weight of tocopherol and 10 parts by weight of deionized water into a water phase pot, and stirring and uniformly dispersing;
and S3, after uniformly stirring, checking the pH value to be 6, and then discharging.
During the preparation steps, the temperature of the water phase pot is controlled at 25 ℃.
The extraction method of the laminaria cuneata extract comprises the following steps:
1) Soaking the laminaria cuneata in warm water of 45 ℃ for 5h, and then cleaning;
2) Crushing the cleaned laminaria cuneata in the step 1) by using a crusher, adding pectinase, protease and cellulase which are 0.9 times the weight of the laminaria cuneata and have the weight ratio of 1;
3) Taking the enzymolysis liquid in the step 2) to inactivate enzyme for 5s at 100 ℃ to obtain an inactivated enzyme liquid;
4) Centrifuging the enzyme inactivating solution obtained in the step 3) at 6000rpm for 10min to obtain a supernatant;
5) Concentrating the supernatant in the step 4), and spray drying to obtain the laminaria cuneata extract.
The extraction method of the sturgeon caviar extract comprises the following steps:
A. crushing sturgeon caviar, and then adding 10 times of treatment liquid by weight to obtain treated sturgeon caviar;
B. cleaning the sturgeon caviar treated in the step A, adding 1 time of deionized water by weight, adding 0.01 time of enzymolysis liquid by weight, and carrying out enzymolysis for 2 hours at 40 ℃ to obtain an enzymolysis product;
C. b, taking the enzymolysis product in the step B, and inactivating the enzyme for 5s at 100 ℃ to obtain an enzyme inactivation solution;
D. centrifuging the enzyme-inactivating liquid obtained in the step C at 6000rpm for 10min to obtain a supernatant;
E. and D, concentrating the supernatant in the step D, and freeze-drying to obtain the sturgeon caviar extract.
In the preparation method of the sturgeon caviar extract, the treatment fluid in the step A comprises 3 parts by weight of sodium chloride, 1 part by weight of hydrogen peroxide, 0.5 part by weight of calcium chloride, 0.5 part by weight of glacial acetic acid and 60 parts by weight of deionized water.
The enzymolysis solution in the step B in the preparation method of the sturgeon caviar extract comprises 1 weight part of papain, 1 weight part of alkaline protease, 1 weight part of lecithinase and 1 weight part of neutral protease.
Comparative example 2
In the comparative example, the weight ratios of the components of each phase are respectively as follows:
0.2 parts by weight of mussel extract, 0.2 parts by weight of grass green salicornia extract, 30 parts by weight of ethanol, 5 parts by weight of thermus thermophilus fermentation product, 10 parts by weight of caprylic/capric triglyceride, 5 parts by weight of VP/VA copolymer, 10 parts by weight of polyethylene glycol-8, 0.2 parts by weight of undaria pinnatifida extract, 2 parts by weight of macadamia mud fermentation product, 2 parts by weight of laminaria saccharina extract, 2 parts by weight of sturgeon caviar extract, 0.01 parts by weight of phenoxyethanol, 0.01 parts by weight of potassium sorbate, 10 parts by weight of glycerin, 0.02 parts by weight of tocopherol, and 10 parts by weight of deionized water.
The Hawaii sea mud fermentation product in the marine organism physical barrier wound protection film comprises 25 parts by weight of sea mud extract, 2 parts by weight of Leuconostoc/radish root fermentation product filtrate and 73 parts by weight of glycerol.
The preparation method of the marine organism physical barrier wound protection film comprises the following steps:
s1, adding 30 parts by weight of ethanol into a water phase pot, slowly adding 5 parts by weight of VP/VA copolymer into the water phase pot, and stirring until the VP/VA copolymer is dissolved uniformly;
s2, sequentially putting 0.2 part by weight of mussel extract, 0.2 part by weight of grass green salicornia extract, 10 parts by weight of polyethylene glycol-8, 5 parts by weight of thermophilic thermus bacteria fermentation product, 10 parts by weight of caprylic/capric triglyceride, 0.2 part by weight of undaria pinnatifida extract, 2 parts by weight of macadamia mud fermentation product, 2 parts by weight of laminaria saccharina extract, 2 parts by weight of sturgeon caviar extract, 0.01 part by weight of phenoxyethanol, 0.01 part by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 part by weight of tocopherol and 10 parts by weight of deionized water into a water phase pot, and stirring and dispersing uniformly;
and S3, after uniformly stirring, checking the pH value to be 6, and then discharging.
During the operation of the preparation steps, the temperature of the water phase pot is controlled at 25 ℃.
The extraction method of the mussel extract comprises the following steps:
(1) Putting 1 weight part of crushed mussel foot silk glands and 10 weight parts of deionized water in an extraction tank for extraction to obtain an extracting solution;
(2) Adding protease 0.02 times the weight of the extract, stirring well, and performing enzymolysis in a water bath kettle at 40 deg.C for 2h to obtain enzymolysis product;
(3) Filtering the enzymolysis product with 200 mesh filter screen, and concentrating the filtrate to obtain mussel extract.
The extraction method of the sturgeon caviar extract comprises the following steps:
A. crushing sturgeon caviar, and then adding 10 times of treatment liquid by weight to obtain treated sturgeon caviar;
B. cleaning the sturgeon caviar treated in the step A, adding 1 time of deionized water by weight, adding 0.01 time of enzymolysis liquid by weight, and carrying out enzymolysis for 2 hours at 40 ℃ to obtain an enzymolysis product;
C. b, taking the enzymolysis product in the step B, and inactivating enzyme for 5s at 100 ℃ to obtain an enzyme inactivation solution;
D. centrifuging the enzyme-inactivating liquid obtained in the step C at 6000rpm for 10min to obtain a supernatant;
E. and D, concentrating the supernatant in the step D, and freeze-drying to obtain the sturgeon caviar extract.
In the preparation method of the sturgeon caviar extract, the treatment fluid in the step A comprises 3 parts by weight of sodium chloride, 1 part by weight of hydrogen peroxide, 0.5 part by weight of calcium chloride, 0.5 part by weight of glacial acetic acid and 60 parts by weight of deionized water.
The enzymolysis solution in the step B in the preparation method of the sturgeon caviar extract comprises 1 weight part of papain, 1 weight part of alkaline protease, 1 weight part of lecithinase and 1 weight part of neutral protease.
Comparative example 3
In the comparative example, the weight ratios of the components of each phase are respectively as follows:
0.2 parts by weight of mussel extract, 0.2 parts by weight of grass green salicornia extract, 30 parts by weight of ethanol, 5 parts by weight of thermus thermophilus fermentation product, 10 parts by weight of caprylic/capric triglyceride, 5 parts by weight of VP/VA copolymer, 2 parts by weight of Laminaria digitata extract, 10 parts by weight of polyethylene glycol-8, 0.2 parts by weight of Undaria pinnatifida extract, 2 parts by weight of Hawaii mud fermentation product, 2 parts by weight of Laminaria saccharina extract, 0.01 parts by weight of phenoxyethanol, 0.01 parts by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 parts by weight of tocopherol, and 10 parts by weight of deionized water.
The Hawaii sea mud fermentation product in the marine organism physical barrier wound protection film comprises 25 parts by weight of sea mud extract, 2 parts by weight of Leuconostoc/radish root fermentation product filtrate and 73 parts by weight of glycerol.
The preparation method of the marine organism physical barrier wound protection film comprises the following steps:
s1, adding 30 parts by weight of ethanol into an aqueous phase pot, then slowly adding 5 parts by weight of VP/VA copolymer into the aqueous phase pot, and stirring until the VP/VA copolymer is uniformly dissolved.
S2, sequentially putting 0.2 part by weight of mussel extract, 0.2 part by weight of salicornia herbacea extract, 10 parts by weight of polyethylene glycol-8, 5 parts by weight of thermophilic thermus thermophilus fermentation product, 2 parts by weight of laminaria cuneata extract, 10 parts by weight of caprylic/capric triglyceride, 0.2 part by weight of undaria pinnatifida extract, 2 parts by weight of macadamia sea mud fermentation product, 2 parts by weight of laminaria saccharina extract, 0.01 part by weight of phenoxyethanol, 0.01 part by weight of potassium sorbate, 10 parts by weight of glycerol, 0.02 part by weight of tocopherol and 10 parts by weight of deionized water into a water phase pot, and uniformly stirring and dispersing.
And S3, after uniformly stirring, checking the pH value to be 6, and then discharging.
During the preparation steps, the temperature of the water phase pot is controlled at 25 ℃.
The extraction method of the mussel extract comprises the following steps:
(1) Putting 1 weight part of crushed mussel foot silk glands and 10 weight parts of deionized water in an extraction tank for extraction to obtain an extracting solution;
(2) Adding protease 0.02 times the weight of the extract, stirring well, and performing enzymolysis in a water bath kettle at 40 deg.C for 2h to obtain enzymolysis product;
(3) Filtering the enzymolysis product with 200 mesh filter screen, and concentrating the filtrate to obtain mussel extract.
The extraction method of the laminaria cuneata extract comprises the following steps:
1) Soaking the laminaria cuneata in warm water at 45 ℃ for 5h, and then cleaning;
2) Crushing the cleaned laminaria cuneata in the step 1) by using a crusher, adding pectinase, protease and cellulase which are 0.9 times the weight of the laminaria cuneata and have the weight ratio of 1;
3) Taking the enzymolysis liquid in the step 2) to inactivate enzyme for 5s at 100 ℃ to obtain an inactivated enzyme liquid;
4) Centrifuging the enzyme inactivating solution obtained in the step 3) at 6000rpm for 10min to obtain a supernatant;
5) Concentrating the supernatant in the step 4), and spray drying to obtain the laminaria cuneata extract.
Wound healing test
The experimental method comprises the following steps: selecting 24 healthy mice, dividing into 4 groups on average, each group comprising 6 mice, each group comprising 3 mice each, removing hair from back with an animal shaver, sterilizing exposed skin with 75% ethanol, and making scald surface with area of 2.5 × 2.5cm with 80 deg.C hot water. The wound protection films prepared in example 1 and comparative examples 1 to 3 were respectively and uniformly applied on the wound surfaces of mice of each group, and were naturally dried. Wound healing rates were recorded on days 3, 8, 13, and 18, respectively. The results are shown in Table 1.
Table 1:
| wound healing Rate (%) | Day 3 | Day 8 | Day 13 | Day 18 |
| Example 1 | >29.2％ | >62.8％ | >82.8％ | >99.7％ |
| Comparative example 1 | >21.3％ | >58.5％ | >79.8％ | >96.6％ |
| Comparative example 2 | >26.8％ | >61.2％ | >80.5％ | >98.9％ |
| Comparative example 3 | >23.6％ | >60.5％ | >81.2％ | >97.5％ |
As can be seen from Table 1, the healing rate of the wound surface is highest in example 1, followed by comparative example 2, followed by comparative example 3, and finally by comparative example 1. The best wound healing effect of example 1 is illustrated.
In addition, pathological examination is carried out on the skin tissues of the drug-coated parts of the experimental mice in the middle stage of the experiment, the ulcer necrosis of each group of mice shows that the lesions reach dermis and subcutaneous edema, and the infiltration degree of subcutaneous lymphocytes is shown in table 2.
Table 2:
as can be seen from Table 2, in example 1, the ulcer and necrosis were not observed and the lesions reached the dermis, and the degree of subcutaneous edema and the degree of subcutaneous lymphocyte infiltration were mild; in comparative example 1, mild ulcer necrosis and lesions reached the dermis, and the degree of subcutaneous edema and subcutaneous lymphocyte infiltration were moderate; in comparative example 2, there was no ulcer necrosis, and lesions were mild in degree of dermal, subcutaneous edema and subcutaneous lymphocyte infiltration; in comparative example 3, mild ulcer necrosis, lesions were infiltrated by dermal and subcutaneous lymphocytes, and the degree of subcutaneous edema was moderate. The best results of example 1 are shown, which are able to repair the skin barrier, promote wound healing and reduce inflammation.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (9)
1. The marine biophysical barrier wound-protecting film is characterized by comprising 0.01-0.2 part by weight of mussel extract, 0.01-0.2 part by weight of turfgrass grass green salt hornwort extract, 30-60 parts by weight of ethanol, 0.01-5 parts by weight of thermus thermophilus fermentation product, 10-30 parts by weight of caprylic acid/capric acid triglyceride, 1-10 parts by weight of VP/VA copolymer, 0.05-1 part by weight of laminaria cuneata extract, 10-30 parts by weight of polyethylene glycol-8, 0.01-0.2 part by weight of undaria pinnatifida extract, 0.01-2 parts by weight of macadamia fermentation product, 0.01-2 parts by weight of laminaria saccharina extract, 0.01-1 part by weight of sturgeon roe extract, 0.01-0.05 part by weight of ethanol phenoxy, 0.01-0.02 part by weight of potassium sorbate, 1-10 parts by weight of glycerol, 0.01-1.02 part by weight of tocopherol and 1-10 parts by weight of deionized water.
2. The marine biophysical barrier wound-protecting film of claim 1, wherein the mussel extract is extracted by a method comprising the steps of:
(1) Putting 1 weight part of crushed mussel foot silk glands and 10 weight parts of deionized water in an extraction tank for extraction to obtain an extracting solution;
(2) Adding protease 0.01-0.05 times of the weight of the extract into the extract, uniformly stirring, and carrying out enzymolysis for 1-2 h in a constant-temperature water bath kettle at 40-50 ℃ to obtain an enzymolysis product;
(3) Filtering the enzymolysis product with 200 mesh filter screen, and concentrating the filtrate to obtain mussel extract.
3. The marine biophysical barrier wound-protecting film according to claim 1, wherein the extraction method of the laminaria cuneata extract comprises the following steps:
1) Soaking the laminaria cuneata in warm water at 45 ℃ for 3-5 h, and then cleaning;
2) Taking the cleaned laminaria cuneata in the step 1), crushing by using a crusher, adding pectinase, protease and cellulase which are 0.9 times the weight of the laminaria cuneata and have the weight ratio of 1;
3) Taking the enzymolysis liquid in the step 2) to inactivate enzyme for 5-8 s at 100-120 ℃ to obtain an inactivated enzyme liquid;
4) Taking the enzyme-inactivating liquid obtained in the step 3) and centrifuging the enzyme-inactivating liquid at 6000 to 8000rpm for 8 to 10min to obtain supernatant;
5) Concentrating the supernatant in the step 4), and spray drying to obtain the laminaria cuneata extract.
4. The marine biophysical barrier wound protecting film according to claim 1, wherein the sturgeon caviar extract is prepared by a method comprising the steps of:
A. crushing sturgeon caviar, and then adding 8-10 times of treatment fluid by weight to obtain treated sturgeon caviar;
B. cleaning the sturgeon caviar treated in the step A, adding 1-2 times of deionized water by weight, adding 0.01 times of enzymolysis liquid by weight, and carrying out enzymolysis at 40-50 ℃ for 2-3 h to obtain an enzymolysis product;
C. b, taking the enzymolysis product in the step B, and inactivating enzyme for 5-8 s at 100-120 ℃ to obtain an enzyme inactivation solution;
D. centrifuging the enzyme-inactivating liquid obtained in the step C at 6000-8000 rpm for 8-10 min to obtain supernatant;
E. and D, concentrating the supernatant in the step D, and freeze-drying to obtain the sturgeon caviar extract.
5. A preparation method of a sturgeon caviar extract according to claim 4, characterized in that the treatment liquid of step A in the preparation method comprises 3-5 parts by weight of sodium chloride, 0.5-1 part by weight of hydrogen peroxide, 0.1-0.5 part by weight of calcium chloride, 0.1-0.5 part by weight of glacial acetic acid and 60-80 parts by weight of deionized water.
6. The method for preparing a sturgeon caviar extract according to claim 4, wherein the enzymatic hydrolysate of step B comprises 1 part by weight of papain, 1 part by weight of alkaline protease, 1 part by weight of lecithinase and 1 part by weight of neutral protease.
7. The marine biophysical barrier wound-protecting film of claim 1, wherein the Hawaii sea mud fermentation product comprises 25 parts by weight of the sea mud extract, 2 parts by weight of the Leuconostoc/radish root fermentation product filtrate, and 73 parts by weight of glycerin.
8. A method for preparing a marine biophysical barrier wound-protecting film according to any one of claims 1 to 7, comprising the steps of:
s1, adding ethanol into a water phase pot, slowly adding the VP/VA copolymer into the water phase pot, and stirring until the VP/VA copolymer is dissolved uniformly;
s2, sequentially putting mussel extract, grass green salicornia extract, polyethylene glycol-8, thermophilic thermus bacterium fermentation product, laminaria cuneata extract, caprylic/capric triglyceride, undaria pinnatifida extract, macadamia mud fermentation product, laminaria saccharina extract, sturgeon caviar extract, phenoxyethanol, potassium sorbate, glycerol, tocopherol and deionized water into a water phase pot, and stirring and dispersing uniformly;
s3, after uniformly stirring, checking the pH value to be 5-6.5, and then discharging.
9. The method for preparing a wound protecting film of a marine biophysical barrier according to claim 8, wherein the temperature of the water phase pot is controlled to be 20-25 ℃.
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