CN114225007B - Whey protein spray for repairing superficial wound surface and preparation method thereof - Google Patents
Whey protein spray for repairing superficial wound surface and preparation method thereof Download PDFInfo
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- CN114225007B CN114225007B CN202111323300.XA CN202111323300A CN114225007B CN 114225007 B CN114225007 B CN 114225007B CN 202111323300 A CN202111323300 A CN 202111323300A CN 114225007 B CN114225007 B CN 114225007B
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- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Classifications
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/12—Carboxylic acids; Salts or anhydrides thereof
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
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- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
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- A61K9/0012—Galenical forms characterised by the site of application
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- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
Abstract
The invention discloses a whey protein spray for repairing superficial wound and a preparation method thereof, wherein the whey protein spray comprises the following components in percentage by mass: whey protein inclusion 1.0% -10.0%, pentanediol 2.0% -3.0%, hyaluronic acid 0.05% -0.5%, dipotassium glycyrrhizinate 0.1% -0.5%, tetrahydropyrimidine carboxylic acid 0.2% -2.0%, madecassoside 0.05% -1%, and the balance purified water, wherein the sum of the mass percentages of the components is 100%. It is also disclosed. The preparation method can maintain the bioactivity of the protein and enhance the stability of the protein by preparing a stable and effective repair preparation which is rich in protein repair active ingredients and can penetrate through skin barriers. By adding external skin preparations, the repairing functional organisms are promoted, the self organism repairing force is effectively improved, and the repairing effect on the superficial wound surface of the skin is obvious.
Description
Technical Field
The invention belongs to the technical field of preparation of medical preparations, and particularly relates to a whey protein spray for repairing superficial wound surfaces and a preparation method of the whey protein spray.
Background
Superficial skin lesions are one of the most common traumatic diseases and may be caused by a variety of factors including mechanical, thermal, optical, biological conditions; superficial damage to the skin, often damaging the epidermis and even affecting the dermis, is caused by various causes. After the skin is damaged, the organism secretes specific regeneration factors to reconstruct the damaged tissue structure; promote proliferation of cells. The skin injury is repaired by relying on the body, the immune system and specific expression factors thereof, the skin barrier reconstruction is often dependent on the repair capability of an individual, but the repair capability of the individual has more influence along with age, nutrition, nursing knowledge level and occasional factors, and wounds which cannot be well repaired are often converted into chronic wounds, persist and repeatedly do not heal, and have great influence on the life and mind and body of a patient.
In clinical treatment, active protein components are externally added outside the wound surface to promote wound repair. Compared with the traditional chemical medicines, the protein medicines have the characteristics of high biological activity, strong specificity and small toxic and side effects. Wound healing is promoted by the use of a wound care formulation rich in growth factors. The growth factors are contained in the wound surface, so that the wound surface has chemotactic effect on epithelial cells, the movement capability of the epithelial cells can be promoted, and the active proteins are applied to provide directionality for migration of the cells, so that the epidermal cells effectively migrate from the wound edge to the center of the wound surface in a directional manner. Under the action of the migration signal, the cells generate power for moving forward as a whole. Through adhesion contact dissociation, cells continuously move forward and continuously migrate from the wound edge to the center of the wound surface, and re-epithelialization of the wound surface is finally completed, so that the wound surface is sealed. By stimulating proliferation and activity of wound surface epithelial cells and fibroblasts; promoting extracellular matrix (ECM) formation, and cytokines acting on many cell types, directly or indirectly increasing the synthesis and secretion of interstitial proteins promote collagen matrix adhesion. Increasing collagen fiber and fibronectin synthesis, thereby causing increased ECM synthesis and deposition, stimulating connective tissue formation.
Whey protein has high application value and potential in accelerating the healing of skin wounds due to the fact that the whey protein is rich in various active ingredients such as Epidermal Growth Factor (EGF), transforming growth factor-a (TGF-a), insulin-like growth factor-1 (IGF-1) and the like. There are extensive literature studies showing that whey proteins can promote wound healing in skin. For example, wang (2010) et al demonstrated that whey peptides are able to promote healing of post-natal wounds in rats by antioxidant and reducing inflammatory responses, etc.; ebaid et al demonstrate that camel milk polypeptides promote wound healing in diabetic rats by coordinating redox status and immune response.
For wound healing, more and more active proteins are used in products for healing skin wound due to the high activity of proteins. However, as a protein substance, the stability in the product is poor, and the protein substance has the defects of easy hydrolysis by protease, short in vivo half-life, low bioavailability and the like. The problem is that the original repair protein with high-efficiency activity cannot fully exert the effect. In addition, for some non-open superficial wounds, the existence of the skin barrier limits the communication between the in-vivo and the in-vitro substances, and the active protein in the product cannot be successfully absorbed through the skin barrier, so that the functions of promoting the organism repairing and wound healing activities are difficult to be fully exerted. In order to increase transdermal absorption and increase the effective active content in use, it is necessary to increase the amount of the material to be fed, which leads to an increase in cost.
Chinese patent (application No. 201310030618.8, publication No. CN 103110935B) discloses skin local hydrolyzed collagen liposome and its preparation method, which provides a hydrolyzed collagen liposome solution. The hydrolyzed collagen liposome prepared from phospholipid, ceramide and cholesterol can promote the hydrolyzed collagen to penetrate through the skin cutin layer and enter the skin, improve the retention of the hydrolyzed collagen in the skin, and can more effectively promote the recovery of skin injury. Chinese patent (application number 200610053301.6, publication number CN 100482212C) provides a preparation method of nano liposome for coating macromolecular medicine, using lecithin and CaCl 2 And macromolecular drugs are prepared by ultrasonic oscillation and standing. The prior inclusion takes lecithin as a shell, and the inclusion made of the lecithin has higher biocompatibility, butThe double-layer film has poor fluidity, hardness, weak flexibility, low skin barrier permeability, high melting point, complex manufacturing process and high cost. Therefore, a stable repair preparation rich in protein repair active ingredients capable of effectively penetrating through skin barrier is developed, so that the biological activity of protein can be maintained, and the stability of the repair preparation can be enhanced. Is necessary for the full repair of superficial wound surface and promoting wound healing.
Disclosure of Invention
The invention aims to provide a whey protein spray for repairing superficial wound surfaces, which solves the problems of poor bioactivity stability and weak penetrating capacity of proteins in the prior art.
The invention adopts the technical scheme that the whey protein spray for repairing superficial wound surface comprises the following components in percentage by mass: whey protein inclusion 1.0% -10.0%, pentanediol 2.0% -3.0%, hyaluronic acid 0.05% -0.5%, dipotassium glycyrrhizinate 0.1% -0.5%, tetrahydropyrimidine carboxylic acid 0.2% -2.0%, madecassoside 0.05% -1%, and the balance purified water, wherein the sum of the mass percentages of the components is 100%.
The present invention is also characterized in that,
the whey protein inclusion is a nano inclusion, and the nano inclusion comprises whey protein, polyalcohol, surfactant, pH buffer component, lipid component and water.
The nano inclusion consists of the following components in percentage by mass: the whey protein comprises 10.0-20.0% of whey protein, 10.0-20.0% of lipid component, 2.0-5.0% of polyalcohol, 5.0-10.0% of surfactant, 0.1-1.0% of pH buffer and the balance of water, wherein the sum of the mass percentages of the components is 100%.
The polyalcohol is one or more of glycerol, propylene glycol, polyethylene glycol 400 and dipropylene glycol; the surfactant is PEG-50 hydrogenated castor oil; the pH buffering agent component is trisodium citrate; the lipid component is cetyl ethyl hexanoate and sorbitan oleate.
The mass ratio of cetyl ethyl hexanoate to sorbitan oleate is 1:3.
the invention adopts another technical scheme that the preparation method of the whey protein spray for repairing superficial wound surface is implemented according to the following steps:
step 1, mixing whey protein, a pH buffer, a polyol and part of water, and stirring to form an aqueous phase system; stirring the lipid component in a water bath at 40-60 ℃ to form an oil phase system; adding the water phase system into the oil phase system at 40-60 ℃, homogenizing for 3-5 times under high pressure of 20000-35000psi, and preserving heat at 40-60 ℃ to obtain W/O inclusion; uniformly mixing water and a surfactant, and preserving heat at 40-60 ℃ to obtain a secondary emulsified water phase system; injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 1-2 times under high pressure of 15000-20000psi to obtain whey protein inclusion;
step 2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 70-80 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, stirring for 30min at 70-80 ℃, cooling to 40-50 ℃, adding whey protein inclusion, and stirring for 20min to obtain the whey protein spray.
Another object of the present invention is to provide a method for preparing the above whey protein spray.
The beneficial effects of the invention are as follows: the preparation method overcomes the defects that the existing repair active protein product is easy to hydrolyze by protease, has short half-life in vivo, low bioavailability and the like by preparing a repair preparation which is stable and effectively penetrates through skin barrier and is rich in protein repair active ingredients. Can not only retain the biological activity of the protein, but also enhance the stability of the protein. By adding external skin preparations, the repairing functional organisms are promoted, the self organism repairing force is effectively improved, and the repairing effect on the superficial wound surface of the skin is obvious.
Detailed Description
The present invention will be described in detail with reference to specific examples.
The invention relates to a whey protein spray for repairing superficial wound surfaces, which comprises the following components in percentage by mass: whey protein inclusion 1.0% -10.0%, pentanediol 2.0% -3.0%, hyaluronic acid 0.05% -0.5%, dipotassium glycyrrhizinate 0.1% -0.5%, tetrahydropyrimidine carboxylic acid 0.2% -2.0%, madecassoside 0.05% -1%, and the balance purified water, wherein the sum of the mass percentages of the components is 100%.
The added pentanediol can promote skin moisturizing, and simultaneously plays a broad-spectrum antibacterial activity, has good inhibition effect on gram positive bacteria, gram negative bacteria, saccharomycetes and mould, and plays a role in bacteriostasis and corrosion prevention.
The added sodium hyaluronate is interwoven into a net shape through a high molecular structure, a layer of breathable film is formed on the surface of the skin, invasion of foreign bacteria and dust is prevented, and the skin is protected from being invaded; improving the living environment of skin cells, reducing nervous system excitability and reducing pain; meanwhile, the wound is sealed, water is prevented from escaping, a strong sealing and humidifying effect is achieved, the degree of wetting of the wound is increased, and a wet healing environment is created for wound healing.
The added tetrahydropyrimidine carboxylic acid plays a role in permeation protection, assists the diffusion and the effect of whey protein inclusion, promotes the long-time exertion of the efficacy and keeps the activity; meanwhile, the characteristics of isotonicity with body fluid are maintained, wound irritation is not caused, good use feeling is provided, and the structure of the bioactive macromolecules is protected from damaging the bioactive macromolecules by the external environment.
The added dipotassium glycyrrhizinate and the hydroxy asiaticoside have anti-inflammatory effect, inhibit inflammation, promote skin repair, and avoid aggravation of inflammation and enhancement of uncomfortable feeling.
The whey protein inclusion is a nano inclusion, and the nano inclusion comprises whey protein, polyalcohol, surfactant, pH buffer component, lipid component and water;
the whey protein comprises 10.0-20.0% of whey protein, 10.0-20.0% of lipid component, 2.0-5.0% of polyalcohol, 5.0-10.0% of surfactant, 0.1-1.0% of pH buffer and the balance of water, wherein the sum of the mass percentages of the components is 100%.
The polyalcohol is one or more of glycerol, propylene glycol, polyethylene glycol 400 and dipropylene glycol; the surfactant is PEG-50 hydrogenated castor oil; the pH buffering agent component is trisodium citrate; the lipid components are cetyl ethyl hexanoate and sorbitan oleate, and the mass ratio of the cetyl ethyl hexanoate to the sorbitan oleate is 1:3, a step of;
the polyols added in the nano inclusion play a role in improving the stability of the whey protein. The polyol forces more water molecules around the protein at low concentrations, thus increasing its stability. The pH buffering component serves to promote protein solubilization and maintain biological activity, and the pH of the final inclusion is maintained in the range of 6.0-7.0 using a pH buffer. The lipid components are sorbitan oleate and cetyl ethyl hexanoate, and the lipid components are homogenized under ultrahigh pressure to form a W/O nano-package shell, so that the possibility of contacting protein with protease is reduced. Double-layer inclusion is formed by re-emulsifying PEG-50 hydrogenated castor oil, and the PEG can improve the thermal stability and prolong the half-life in vivo after being wrapped.
The invention discloses a preparation method of whey protein spray for repairing superficial wound surfaces, which is implemented according to the following steps:
step 1, preparing whey protein inclusion, specifically:
step 1.1, weighing according to mass concentration percentage: the whey protein comprises 10.0-20.0% of whey protein, 10.0-20.0% of lipid component, 2.0-5.0% of polyalcohol, 5.0-10.0% of surfactant, 0.1-1.0% of pH buffer and the balance of water, wherein the sum of the mass percentages of the components is 100%;
step 1.2, mixing whey protein, a pH buffer, a polyol and water, stirring and dissolving to form a water phase system, wherein the water content is 10.0% -20.0%;
step 1.3, stirring and dissolving lipid components in a constant-temperature water bath at 40-60 ℃ to form an oil phase system;
step 1.4, adding the water phase system into the oil phase system at 40-60 ℃, homogenizing for 3-5 times at 20000-35000psi under high pressure, and then preserving heat at 40-60 ℃ to obtain W/O inclusion;
step 1.5, weighing the rest water and the surfactant, uniformly mixing, and preserving heat at 40-60 ℃ to obtain a secondary emulsified water phase system;
step 1.6, injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 1-2 times under high pressure of 15000-20000psi to obtain whey protein inclusion;
step 2, preparing whey protein spray, which specifically comprises the following steps:
step 2.1, weighing according to mass percent: whey protein inclusion 1.0% -10.0%, pentanediol 2.0% -3.0%, hyaluronic acid 0.05% -0.5%, dipotassium glycyrrhizinate 0.1% -0.5%, tetrahydropyrimidine carboxylic acid 0.2% -2.0%, madecassoside 0.05% -1%, and the balance purified water, wherein the sum of the mass percentages of the components is 100%;
step 2.2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 70-80 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, and stirring for 30min at 70-80 ℃ at a stirring speed of 250-500 rpm;
step 2.3, cooling to 40-50 ℃ after the step 2.2, adding the whey protein inclusion, stirring for 20min, wherein the stirring speed is 250-300 rpm, and obtaining the whey protein spray.
Example 1
The invention discloses a preparation method of whey protein spray for repairing superficial wound surfaces, which is implemented according to the following steps:
step 1, preparing whey protein inclusion, specifically:
step 1.1, weighing according to mass concentration percentage: the mass percentage concentration of the whey protein is 10.0%, the mass percentage concentration of cetyl ethyl hexanoate is 2.5%, the mass percentage concentration of sorbitan oleate is 7.5%, the mass percentage concentration of glycerin is 2.0%, the mass percentage concentration of the surfactant is 5.0%, the mass percentage concentration of trisodium citrate is 0.5%, and the balance is water, wherein the sum of the mass percentages of the components is 100%;
step 1.2, mixing whey protein, trisodium citrate and glycerin with water, stirring and dissolving to form a water phase system, wherein the water content is 20.0%;
step 1.3, stirring and dissolving cetyl ethyl hexanoate and sorbitan oleate in a constant-temperature water bath at 40 ℃ to form an oil phase system;
step 1.4, adding the water phase system into the oil phase system at 40 ℃, homogenizing for 3 times at 35000psi under high pressure, and then preserving heat at 40 ℃ to obtain W/O inclusion;
step 1.5, weighing the rest water and PEG-50 hydrogenated castor oil, uniformly mixing, and preserving heat at 40 ℃ to obtain a secondary emulsified water phase system;
step 1.6, injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 2 times under high pressure at 15000psi to obtain whey protein inclusion;
step 2, preparing whey protein spray, which specifically comprises the following steps:
step 2.1, weighing according to mass percent: 10.0% of whey protein inclusion, 3.0% of pentanediol, 0.05% of hyaluronic acid, 0.5% of dipotassium glycyrrhizinate, 2.0% of tetrahydropyrimidine carboxylic acid, 0.05% of madecassoside and the balance of purified water, wherein the sum of the mass percentages of the components is 100%;
step 2.2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 80 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, and stirring for 30min at 80 ℃ at a stirring speed of 500rpm;
step 2.3, cooling to 40 ℃ after the step 2.2, adding the whey protein inclusion, stirring for 20min at the stirring speed of 250rpm, and obtaining the whey protein spray.
Example 2
The invention discloses a preparation method of whey protein spray for repairing superficial wound surfaces, which is implemented according to the following steps:
step 1, preparing whey protein inclusion, specifically:
step 1.1, weighing according to mass concentration percentage: 20.0% of whey protein, 5% of cetyl ethyl hexanoate, 15% of sorbitan oleate, 5.0% of propylene glycol, 10.0% of PEG-50 hydrogenated castor oil, 0.5% of trisodium citrate, and the balance of water, wherein the sum of the mass percentages of the components is 100%;
step 1.2, mixing whey protein, trisodium citrate and propylene glycol with water, stirring and dissolving to form a water phase system, wherein the water content is 10.0%;
step 1.3, stirring and dissolving cetyl ethyl hexanoate and sorbitan oleate in a constant-temperature water bath at 60 ℃ to form an oil phase system;
step 1.4, adding the water phase system into the oil phase system at 60 ℃, homogenizing for 5 times under 20000psi at high pressure, and then preserving heat at 60 ℃ to obtain W/O inclusion;
step 1.5, weighing the rest water and PEG-50 hydrogenated castor oil, uniformly mixing, and preserving heat at 60 ℃ to obtain a secondary emulsified water phase system;
step 1.6, injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 1 time under high pressure of 20000psi to obtain whey protein inclusion;
step 2, preparing whey protein spray, which specifically comprises the following steps:
step 2.1, weighing according to mass percent: whey protein inclusion 1.0%, pentanediol 2.0%, hyaluronic acid 0.5%, dipotassium glycyrrhizinate 0.1%, tetrahydropyrimidine carboxylic acid 0.2%, madecassoside 1%, and purified water in balance, wherein the sum of the mass percentages of the components is 100%;
step 2.2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 70 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, and stirring for 30min at 70 ℃ at a stirring speed of 250rpm;
step 2.3, cooling to 50 ℃ after the step 2.2, adding the whey protein inclusion, stirring for 20min at the stirring speed of 300rpm, and obtaining the whey protein spray.
Example 3
The invention discloses a preparation method of whey protein spray for repairing superficial wound surfaces, which is implemented according to the following steps:
step 1, preparing whey protein inclusion, specifically:
step 1.1, weighing according to mass concentration percentage: 15% of whey protein, 5% of cetyl ethyl hexanoate, 15% of sorbitan oleate, 4.0% of polyethylene glycol 400, 8% of PEG-50 hydrogenated castor oil, 0.1% of trisodium citrate and the balance of water, wherein the sum of the mass percentages of the components is 100%;
step 1.2, mixing whey protein, PEG-50 hydrogenated castor oil and polyethylene glycol 400 with water, stirring and dissolving to form a water phase system, wherein the water content is 15%;
step 1.3, stirring and dissolving cetyl alcohol ethyl hexanoate and sorbitan oleate in a constant-temperature water bath at 50 ℃ to form an oil phase system;
step 1.4, adding the water phase system into the oil phase system at 40-60 ℃, homogenizing for 4 times at 30000psi under high pressure, and then preserving heat at 50 ℃ to obtain a W/O inclusion;
step 1.5, weighing the rest water and surfactant, uniformly mixing, and preserving heat at 50 ℃ to obtain a secondary emulsified water phase system;
step 1.6, injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 2 times under high pressure at 15000psi to obtain whey protein inclusion;
step 2, preparing whey protein spray, which specifically comprises the following steps:
step 2.1, weighing according to mass percent: whey protein inclusion 5%, pentanediol 2.5%, hyaluronic acid 0.1%, dipotassium glycyrrhizinate 0.2%, tetrahydropyrimidine carboxylic acid 1.0%, madecassoside 0.3%, and purified water in balance, wherein the sum of the mass percentages of the components is 100%;
step 2.2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 75 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, and stirring for 30min at 75 ℃ at a stirring speed of 350rpm;
step 2.3, cooling to 40 ℃ after the step 2.2, adding the whey protein inclusion, stirring for 20min at the stirring speed of 250rpm, and obtaining the whey protein spray.
Example 4
The invention discloses a preparation method of whey protein spray for repairing superficial wound surfaces, which is implemented according to the following steps:
step 1, preparing whey protein inclusion, specifically:
step 1.1, weighing according to mass concentration percentage: 15% of whey protein, 3.75% of cetyl ethyl hexanoate, 11.25% of sorbitan oleate, 5.0% of dipropylene glycol, 10.0% of PEG-50 hydrogenated castor oil, 0.5% of trisodium citrate, and the balance of water, wherein the sum of the mass percentages of the components is 100%;
step 1.2, mixing whey protein, trisodium citrate and dipropylene glycol with water, stirring and dissolving to form a water phase system, wherein the water content is 15%;
step 1.3, stirring and dissolving cetyl alcohol ethyl hexanoate and sorbitan oleate in a constant-temperature water bath at 50 ℃ to form an oil phase system;
step 1.4, adding the water phase system into the oil phase system at 50 ℃, homogenizing for 4 times under 20000psi at high pressure, and then preserving heat at 50 ℃ to obtain W/O inclusion;
step 1.5, weighing the rest water and surfactant, uniformly mixing, and preserving heat at 50 ℃ to obtain a secondary emulsified water phase system;
step 1.6, injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 1 time under high pressure at 15000psi to obtain whey protein inclusion;
step 2, preparing whey protein spray, which specifically comprises the following steps:
step 2.1, weighing according to mass percent: 8% of whey protein inclusion, 2.5% of pentanediol, 0.2% of hyaluronic acid, 0.3% of dipotassium glycyrrhizinate, 1.5% of tetrahydropyrimidine carboxylic acid, 0.8% of madecassoside and the balance of purified water, wherein the sum of the mass percentages of the components is 100%;
step 2.2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 80 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, and stirring for 30min at 80 ℃ at a stirring speed of 400rpm;
step 2.3, cooling to 40 ℃ after the step 2.2, adding the whey protein inclusion, stirring for 20min at the stirring speed of 250rpm, and obtaining the whey protein spray.
The whey protein spray of the invention is smeared on the superficial wound surface of the skin, thereby verifying the repairing effect of the composition for (repairing) the superficial wound surface. The commercial repair spray like products are used as the reference substances. Verification was performed using the product of example 4.
1. Rabbit wound repair test
The effectiveness was evaluated by applying a material to rabbit skin by pouring, and evaluating its safety by gross visual and histological observation, using collagen staining. The safety and effectiveness of the test are evaluated by comparison with a reference.
1.1 moulding and preparation method
The backs of rabbits were dehaired and sterilized. Selecting a back area with the size of 4 blocks of 2cm by 2cm for making wounds, smearing 0.3-0.4ml of test product in the selected area, using a traditional microneedle with the size of 0.5mm after smearing, perpendicularly crossing 4 directions for treatment, rolling for 5 times in each direction, and repeating the treatment for 4 times, wherein the treatment is suitable for slight redness, swelling and micro bleeding of skin.
After the trauma treatment is completed, the selected area is treated4cm 2 ) Respectively smearing a test group and a control group on two parts of the test group and the control group, wherein one part is used as a blank control area; the application areas are as far apart as possible from each other and do not interfere with each other. Observing the appearance change of the skin at the application position, side effects and the like.
Coating amount and period: each time, 0.4ml each time, is applied for 14 days.
1.2 evaluation of safety
1.2.1 general observations: the appearance change, side effects and wound healing conditions of the smearing position are observed regularly, wherein the side effects comprise erythema, swelling and the like, the wound size and the red swelling fading conditions are mainly erythema and swelling, and the skin irritation response is scored, photographed and recorded.
When the experimental result is molded by using the traditional microneedle (0.5 mm), the skin preparation area of the animal has the phenomenon of reddening, and part of the skin preparation area has the phenomenon of microhemorrhage, the statistical result is shown in the following table 1, and the specific observation record is shown in the following table 2:
TABLE 1 general observations after wound creation
| Observation of | |
| 3 days after wound making | The blank area is seen as micro bleeding; test group, control see reddish |
| 7 days after wound making | The blank area is seen as micro bleeding; test group, control see reddish |
| 14 days after wound making | All see the phenomenon of reddening |
Table 2 table of symptom observation records
1.2.2 histological examination: 3 samples were sacrificed on day 3, day 7, and day 14, the residual condition of the samples was checked, the intradermal tissues around the injection site were removed, 4% paraformaldehyde fixed, paraffin embedded, sectioned (5 μm), HE stained, and photographed by light microscopy.
As a result of experiments, the HE staining observation shows that the inflammatory cells are rare, the test product and the control product have no irritation, and the control product has slightly higher relative irritation; the specific results are shown in Table 3:
TABLE 3 HE staining pathology classification and classification statistics
Evaluation of validity
On the 3 rd day, 7 th day and 14 th day, animals are sacrificed, materials and surrounding intradermal tissues are taken out, 4% paraformaldehyde is fixed, paraffin is embedded, the slices are cut, the slices are subjected to dyeing observation by a picric acid sirius scarlet dyeing method, the thickness of the slices is 6um, water is dewaxed conventionally, and the slices are placed in a 0.1% picric acid sirius scarlet dyeing solution for dyeing for 1 hour, and the slices are washed 3 times by double distilled water; mayer hematoxylin counterstain for 15 minutes, and double distilled water washes for 3 times; dehydrating, transparent, sealing, observing under a polarized light microscope, and photographing. The degree of damage and repair of skin tissue was observed.
The sirius red staining results are shown in table 4, collagen fiber content for each group: test article > control article > blank control article. On the repair results, both the test product and the control group have the effect of repairing the damage, and the embodiment can obviously improve the damage condition of the model skin, has the help effect on nursing superficial wound surfaces or surrounding skin, and has the equivalent efficacy on similar products.
TABLE 4 Langxing red dyeing collagen fiber fractionation statistics
2. Human body needle wound repair test
The inflammation, appearance and wound healing of the red areas of the subjects before and after needle roller therapy and using the skin external agent care system were analyzed by a visual image analyzer.
The verification scheme is as follows: selecting 10 patients subjected to medical treatment such as microneedle water exposure, and the like, and continuously using for 1 week after treatment, 4 times per day, wherein each half face is 0.5ml (4 spray), wherein the left side is a test group, and the right side is a control group;
the patient before and after treatment was photographed using a visual skin detector (Canfield company, usa), and the inflammation of the skin was scored and judged by the red area value. While doctor diagnosis and treatment judged that on the fifth day, examination was performed, more significant epithelialization, less skinning and reduced swelling were observed on the left side of the face than on day 2. As shown in table 5, the right side of the face treated with the comparative composition showed less epithelialization, more retained skinning, and slower healing.
In addition, subjective reports perceived less irritation, discomfort, dryness, and itching on the left side of their face compared to the right side.
TABLE 5 Visia skin Detector Red area detection values
The improvement rate of the red zone of the test group is 70 percent; the improvement rate of the red zone of the control group accounts for 40%; the "red zone" improvement rate test group was more remarkable than the control group.
The whey protein spray for repairing superficial wound surface has good protein stability, excellent biocompatibility and moisture retention performance. The product has no toxicity, no irritation, no sensitization, and high safety; is easy to be absorbed by skin; compared with the prior nano-wrapping, the absorption rate after nano-wrapping is higher, and the repairing effect of the superficial wound surface of the skin is obvious.
Claims (2)
1. The whey protein spray for repairing the superficial wound surface is characterized by comprising the following components in percentage by mass: whey protein inclusion 1.0% -10.0%, pentanediol 2.0% -3.0%, hyaluronic acid 0.05% -0.5%, dipotassium glycyrrhizinate 0.1% -0.5%, tetrahydropyrimidine carboxylic acid 0.2% -2.0%, madecassoside 0.05% -1%, and the balance purified water, wherein the sum of the mass percentages of the components is 100%;
the whey protein inclusion is a nano inclusion, and the nano inclusion comprises whey protein, polyalcohol, surfactant, pH buffer component, lipid component and water; the nano inclusion consists of the following components in percentage by mass: the whey protein comprises 10.0-20.0% of whey protein, 10.0-20.0% of lipid component, 2.0-5.0% of polyalcohol, 5.0-10.0% of surfactant, 0.1-1.0% of pH buffer and the balance of water, wherein the sum of the mass percentages of the components is 100%;
the polyalcohol is one or more of glycerol, propylene glycol, polyethylene glycol 400 and dipropylene glycol; the surfactant is PEG-50 hydrogenated castor oil; the pH buffering agent component is trisodium citrate; the lipid component is cetyl alcohol ethyl caproate and sorbitan oleate;
the preparation method of the whey protein spray for repairing the superficial wound surface is implemented according to the following steps:
step 1, mixing whey protein, a pH buffer, a polyol and part of water, and stirring to form an aqueous phase system; stirring the lipid component in a water bath at 40-60 ℃ to form an oil phase system; adding the water phase system into the oil phase system at 40-60 ℃, homogenizing for 3-5 times under high pressure of 20000-35000psi, and preserving heat at 40-60 ℃ to obtain a W/O inclusion; uniformly mixing water and a surfactant, and preserving heat at 40-60 ℃ to obtain a secondary emulsified water phase system; injecting the W/O inclusion into a secondary emulsified water phase system, and homogenizing for 1-2 times under high pressure of 15000-20000psi to obtain whey protein inclusion;
and 2, stirring and dissolving dipotassium glycyrrhizinate, tetrahydropyrimidine carboxylic acid, madecassoside and water in a constant-temperature water bath at 70-80 ℃, then adding evenly dispersed pentanediol and hyaluronic acid, stirring for 30min at 70-80 ℃, cooling to 40-50 ℃, adding whey protein inclusion, and stirring for 20min to obtain the whey protein spray.
2. The whey protein spray for repairing superficial wound surface according to claim 1, wherein the mass ratio of cetyl ethyl hexanoate to sorbitan oleate is 1:3.
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