CN115177535A - Deer skin collagen polypeptide liposome hydrogel, preparation method and application thereof - Google Patents

Abstract

The embodiment of the application belongs to the technical field of biology, and relates to a deer skin collagen polypeptide liposome hydrogel, a preparation method and an application thereof, wherein the deer skin collagen polypeptide liposome hydrogel is prepared from 10-50 g of deer skin collagen polypeptide, 20-100 g of egg yolk lecithin, 10-20 g of cholesterol, 2-5 g of tween-80, 2-10 g of triethanolamine, 2-10 g of carbomer and 100ml of chloroform according to per 100ml of solution. The deer skin collagen polypeptide liposome hydrogel can effectively delay skin aging.

Description

Deer skin collagen polypeptide liposome hydrogel, preparation method and application thereof

Technical Field

The application relates to the technical field of biology, in particular to a deer skin collagen polypeptide liposome hydrogel, a preparation method and application thereof.

Background

Skin aging is the result of a combination of intrinsic and extrinsic factors. Intrinsic aging involves intrinsic factors related to age, in terms of neuroendocrine, while extrinsic aging is the result of external factors, such as ultraviolet a (UVA) and ultraviolet B (UVB) radiation exposure, environmental pollution and the production of reactive oxygen species. The signs of ultraviolet-induced skin aging, also known as photoaging, include deep wrinkles, loss of elasticity, increased dryness and roughness, telangiectasia, and irregular pigmentation. The presence of UVB in solar uv light can directly damage DNA, resulting in most of the light aging. Ultraviolet radiation-induced Reactive Oxygen Species (ROS) play an important role in photoaging. In addition to directly attacking macromolecules such as proteins, lipids, RNA and DNA, the excess reactive oxygen species in the skin trigger a series of signal transduction, down-regulating collagen (the major component in the skin) biosynthesis, and up-regulating a series of Matrix Metalloproteinases (MMPs), leading to collagen degradation. These changes in the skin can lead to a photoaging phenotype.

In recent years, the use of antioxidants as functional ingredients in the human diet has increased dramatically. One popular ingredient that is considered an antioxidant is collagen in hydrolyzed form. Collagen is a very desirable fibrin in the biomedical and cosmetic industries because it is beneficial to the skin, biocompatible, bioactive, and poorly immunogenic. This biomaterial consists of repeated triplets of the amino acids glycine (Gly), proline (Pro) and hydroxyproline (Hyp). Collagen can be isolated from a variety of sources, resulting in differences in its chemical and thermal properties. Proteins in their native form, such as collagen, exhibit low solubility in water due to their high molecular weight of approximately 300 kDa. These natural characteristics present difficulties for their use in cosmetic and edible products. However, degradation of collagen produces Hydrolyzed Collagen (HC), which consists of fragment proteins with low molecular weights (Mw) between 1kDa and 10 kDa. HC is obtained by denaturing native collagen, then enzymatically breaking down the protein chain into small peptides, cleaving the protein at specific amide bonds. Natural peptides (e.g., collagen hydrolysates) are widely used due to their excellent biocompatibility, biodegradability and weak antigenicity.

Skin aging is classified into natural aging and photoaging (extrinsic aging). The increase in age leads to a gradual loss and rupture of the dermal collagen fibers of the human skin, and the total collagen content is lost at a rate of 1% per year over 25 years, resulting in atrophy and collapse of the skin tissue, manifested by thinning, loss of elasticity, dryness, roughness, and eventual formation of wrinkles. The main causes of two clinical changes in aging skin are: collagen (the major structural component of the skin). Talwar et al found that the levels of type I and type III procollagens were significantly lower than that of severely photodamaged human skin. Overall, in vivo, collagen synthesis in photoaged human skin is reduced more than in naturally aged skin. Exogenous aging is caused by changes in the external environment (wind, sun, chemical corrosion), irregular living habits (smoking and drinking), and the like. In 1986, "Photoaging" (Photoaging) was first defined as a clinical result of chronic lesions accumulated in the skin due to the long-term exposure to Ultraviolet (UV) radiation. The main causes of Skin Photoaging (SP) are long-term uv irradiation, and the increase of accumulation degree increases melanin content in the Skin, directly or indirectly damages nuclear DNA, activates related cell receptors in the Skin epidermis and dermis, and finally leads to collagen fragmentation and stop synthesizing new collagen. Collagen in the dermis layer is basically not repaired after being broken, so that the structurally intact skin has certain defects, scars are formed, and the skin is finally atrophied and wrinkles are formed. The cumulative ultraviolet radiation induced photoaging of the skin is clinically common as: redness and dryness, skin breakdown, muscle relaxation, rough wrinkles, abnormal local pigmentation, and a leather-like appearance, with deeper wrinkles being most typical.

With the improvement of the knowledge of people on cosmetics, the production of bioactive peptides by using natural resources has not been able to meet the increasing demand of people. Thus, some researchers have utilized biotechnology to synthesize biologically active peptides. On the one hand, this may improve the efficiency of production of biologically active peptides. On the other hand, researchers have used various methods to compensate for the problems of poor stability and poor skin permeability, such as peptide structure modification, peptide molecular weight adjustment, or derivatization methods. However, there is still a lack of a drug which can prolong the residence time on the skin and is effective in delaying skin aging.

Disclosure of Invention

The embodiment of the application aims to provide a deer skin collagen polypeptide liposome hydrogel, a preparation method and application thereof, which can effectively delay skin aging.

In order to solve the above technical problems, the embodiment of the present application provides a deer skin collagen polypeptide liposome hydrogel, which adopts the following technical scheme:

the deer skin collagen polypeptide liposome hydrogel is prepared from every 100ml of solution, and comprises the following components:

10 to 50g of deer skin collagen polypeptide, 20 to 100g of yolk lecithin, 10 to 20g of cholesterol, 2 to 5g of tween-80, 2 to 10g of triethanolamine, 2 to 10g of carbomer and 100ml of chloroform.

Further, the deer skin collagen polypeptide liposome hydrogel is prepared by mixing the components in each 100ml of solution, and comprises the following components:

10g of deer skin collagen polypeptide, 20g of yolk lecithin, 10g of cholesterol, 2g of tween-80, 2g of triethanolamine, 2-10 g of carbomer and 100ml of chloroform.

In order to solve the above technical problems, an embodiment of the present application further provides a preparation method of the deer skin collagen polypeptide liposome hydrogel, which adopts the following technical scheme:

preparing deer skin collagen polypeptide;

dissolving egg yolk lecithin, cholesterol and tween-80 in a chloroform solvent to obtain a mixed solution;

spin-drying the mixed solution to transparent film shape to obtain liposome, adding solution containing deer skin collagen polypeptide into the liposome, and hydrating to obtain deer skin collagen polypeptide;

placing the deer skin collagen polypeptide in a centrifuge tube, oscillating, performing ultrasound by using an ice water bath probe to obtain uniformly dispersed deer skin collagen polypeptide liposome;

uniformly mixing carbomer weighed in advance with ultrapure water, standing overnight, and adding triethanolamine to obtain blank hydrogel;

and adding the deer skin collagen polypeptide liposome into the blank hydrogel to obtain the collagen polypeptide liposome hydrogel.

Further, the step of preparing deer skin collagen polypeptide comprises:

preparing deer skin collagen;

adding alkaline protease into the deer skin collagen for hydrolysis reaction, and inactivating enzyme in boiling water after 4h to obtain hydrolysate;

and cooling and centrifuging the hydrolysate, taking supernatant, and freeze-drying to obtain the dried deer skin collagen polypeptide.

Further, the step of preparing deer skin collagen comprises:

dissolving deer skin glue powder in acetic acid water solution, magnetically stirring, and filtering to obtain filtrate;

centrifuging the filtrate, and taking a supernatant, wherein the supernatant is a crude acid-soluble collagen solution;

salting out the acid-soluble crude collagen solution to obtain a crude collagen product;

dissolving the collagen crude product in acetic acid solution, dialyzing in 0.1M acetic acid solution for 48 hr, dialyzing in ultrapure water for 24 hr, and freeze-drying to obtain the deer skin collagen.

Further, the step of salting out the acid-soluble crude collagen solution to obtain a crude collagen product comprises:

and adding sodium chloride into the acid-soluble crude collagen solution until the final concentration of the sodium chloride is 2.0mol/L, centrifuging, and retaining a precipitate to obtain the crude collagen product.

Further, the concentration of the acetic acid aqueous solution is 0.5mol/L.

Further, the mass ratio of the deerskin glue powder to the acetic acid aqueous solution is 1.

Further, the molar ratio of the egg yolk lecithin to the cholesterol is 3.

Further, the step of spin-drying the mixed solution to a transparent film shape comprises:

and slowly vacuumizing the mixed solution at the rotation speed of 90rmp, carrying out water bath at the temperature of 30 ℃, and spin-drying the mixed solution to form a transparent film.

In order to solve the technical problems, the embodiment of the application also provides an application of the deer skin collagen polypeptide liposome hydrogel in improving skin cosmetic components.

Compared with the prior art, the embodiment of the application mainly has the following beneficial effects:

the deer skin collagen polypeptide is wrapped in the liposome to prepare hydrogel, so that the physical stability, the chemical stability and the like of the polypeptide are improved. The use of liposomes as carriers herein can improve penetration of drugs through the skin by controlling drug release, increasing the residence time of the drug on the skin, allowing direct contact with the stratum corneum or skin appendages, and reducing the physicochemical instability of the drug.

Drawings

In order to more clearly illustrate the solution of the present application, the drawings needed for describing the embodiments of the present application will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present application, and that other drawings can be obtained by those skilled in the art without inventive effort.

Fig. 1 is a flow diagram of one embodiment of a method for preparing a deer skin collagen polypeptide liposome hydrogel according to the present application;

FIG. 2 is a pictorial view of a product according to one embodiment of a deer skin collagen polypeptide liposome hydrogel of the present application;

FIG. 3 is a graph of sustained release of one embodiment of a method of preparing a deer skin collagen polypeptide liposome hydrogel according to the present application;

FIG. 4 is a scanning electron microscope image of a deer skin collagen polypeptide liposome hydrogel according to the present application;

fig. 5 is a graph showing the result of a skin permeation experiment according to an embodiment of a method for preparing a deer skin collagen polypeptide liposome hydrogel according to the present application;

fig. 6 is a graph showing content stability results of one example of a method for preparing a deer skin collagen polypeptide liposome hydrogel according to the present application.

Fig. 7 is a graph showing the results of changes in the skins of UVB-irradiated mice by deer skin collagen polypeptide liposome hydrogel according to the present application.

Fig. 8 is a graph of the results of physical stability of a deer skin collagen polypeptide liposome hydrogel according to the present application.

Fig. 9 is a graph of the results of the heat and cold resistant centrifugal stability of the deer skin collagen polypeptide liposome hydrogel according to the present application.

Detailed Description

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs; the terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application; the terms "including" and "having," and any variations thereof, in the description and claims of this application and the description of the above figures are intended to cover non-exclusive inclusions. The terms "first," "second," and the like in the description and claims of this application or in the above-described drawings are used for distinguishing between different objects and not for describing a particular order.

Reference herein to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the application. The appearances of the phrase in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. It is explicitly and implicitly understood by one skilled in the art that the embodiments described herein may be combined with other embodiments.

The following examples are provided to facilitate a better understanding of the present application and are not intended to limit the same. The experimental procedures in the following examples are all conventional ones unless otherwise specified. The experimental materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.

In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings.

1. A deer skin collagen polypeptide liposome hydrogel, as shown in fig. 2 and 4, fig. 2 is a product entity diagram of an embodiment of the deer skin collagen polypeptide liposome hydrogel according to the present application. Fig. 4 is a scanning electron microscope result image of deer skin collagen polypeptide liposome hydrogel according to the present application. Wherein, part A in FIG. 4 represents DCP-LIP-Gel (deer skin collagen polypeptide liposome hydrogel) structure at 50 μm; part B of FIG. 4 shows the structure of DCP-LIP-Gel (deer skin collagen polypeptide liposome hydrogel) at 2 μm. The deer skin collagen polypeptide liposome hydrogel is prepared from 100ml of solution, and comprises the following components:

10-50 g of deer skin collagen polypeptide, 20-100 g of egg yolk lecithin, 10-20 g of cholesterol, 2-5 g of tween-80, 2-10 g of triethanolamine, 2-10 g of carbomer and 100ml of chloroform.

Prepared according to the prescription amount of 100ml, and is specifically shown in the following table 1:

in addition, the present application also provides the following three embodiments:

example 1: the deer skin collagen polypeptide liposome hydrogel is prepared from 10g of deer skin collagen polypeptide, 20g of egg yolk lecithin, 10g of cholesterol, 2g of tween-80, 2g of triethanolamine, 2g of carbomer and 100ml of chloroform according to the proportion of each 100ml of solution.

Example 2: the deer skin collagen polypeptide liposome hydrogel is prepared from 30g of deer skin collagen polypeptide, 50g of egg yolk lecithin, 15g of cholesterol, 3g of tween-80, 5g of triethanolamine, 5g of carbomer and 100ml of chloroform according to the proportion of each 100ml of solution.

Example 3: the deer skin collagen polypeptide liposome hydrogel is prepared from 50g of deer skin collagen polypeptide, 100g of yolk lecithin, 20g of cholesterol, 5g of tween-80, 10g of triethanolamine, 10g of carbomer and 100ml of chloroform according to the proportion of each 100ml of solution.

2. As shown in fig. 1, fig. 1 is a flow chart of an embodiment of a method for preparing a deer skin collagen polypeptide liposome hydrogel according to the present application. A preparation method of deer skin collagen polypeptide liposome hydrogel comprises the following steps:

s1: preparing deer skin collagen polypeptide;

s2: dissolving egg yolk lecithin, cholesterol and tween-80 in a chloroform solution to obtain a mixed solution;

s3: spin-drying the mixed solution to transparent film shape to obtain liposome, adding solution containing deer skin collagen polypeptide into the liposome, and hydrating to obtain deer skin collagen polypeptide liposome;

s4: placing the deer skin collagen polypeptide liposome in a centrifuge tube, oscillating the centrifuge tube, performing ultrasonic treatment with an ice-water bath probe to obtain the deer skin collagen polypeptide liposome which is uniformly dispersed;

s5: uniformly mixing carbomer weighed in advance with ultrapure water, standing overnight, and adding triethanolamine to obtain blank hydrogel;

s6: and adding the deer skin collagen polypeptide liposome into the blank hydrogel to obtain the collagen polypeptide liposome hydrogel.

The preparation process of the deer skin collagen polypeptide comprises the following steps:

dissolving deer skin gelatin powder in acetic acid aqueous solution with the concentration of 0.5mol/L, magnetically stirring at 4 ℃ for 48 hours, and filtering to obtain filtrate, wherein the mass ratio of the deer skin gelatin powder to the acetic acid aqueous solution is 1.

Centrifuging the filtrate at 8000r/min for 30min, and collecting supernatant, wherein the supernatant is crude solution of acid soluble collagen;

salting out sodium chloride (NaCl is added between 0.8M and 2.6M), controlling the final concentration of the sodium chloride to be 2.0mol/L, centrifuging at 4000rpm for 15min, removing supernatant, and collecting precipitate to obtain a collagen crude product;

dissolving the crude collagen product in a small amount of 0.5mol/L acetic acid solution, dialyzing in 0.1M acetic acid solution for 48h, dialyzing in ultrapure water for 24h, pouring the dialyzed crude collagen product into a watch glass, freezing at-80 ℃ for 24h in a refrigerator, and freeze-drying in a freeze dryer for 24h to obtain deer skin collagen (DC).

Preparing deer skin collagen into a solution with a certain concentration, preheating at 90 ℃ to depolymerize the deer skin collagen, cooling to a certain temperature, adding alkaline protease according to the enzyme-substrate ratio of 6000U/g to start a hydrolysis reaction at 55 ℃ and pH 8.0, adjusting the pH value to 8.0 by using 1.0mol/L NaOH or 1.0mol/L HCl every half hour in the reaction process to keep the pH value constant, and after hydrolyzing for 4 hours, inactivating the enzyme by using boiling water to obtain a hydrolysate;

cooling, centrifuging the hydrolysate at 4000r/min for 15min, freeze drying the supernatant to obtain deer skin collagen polypeptide (DCP), and storing at-20 deg.C.

The preparation method of the collagen polypeptide liposome hydrogel according to the deer skin collagen polypeptide comprises the following specific steps:

weighing a proper amount of egg yolk lecithin and cholesterol according to a molar ratio of 3;

slowly vacuumizing the mixed solution at the rotation speed of 90rpm, controlling the water bath temperature to be 30 ℃, spin-drying the solvent to enable the mixed solution to be in a transparent film shape, adding the solution containing the deer skin collagen polypeptide, and hydrating for 2 hours at 37 ℃ to obtain deer skin collagen polypeptide liposome;

placing the hydrated deer skin collagen polypeptide liposome in a centrifuge tube, shaking for vortex, and performing ultrasonic treatment with a probe in ice water bath for 10min to obtain uniformly dispersed deer skin collagen polypeptide liposome;

weighing carbomer and ultrapure water according to the formula, putting the carbomer and the ultrapure water into a beaker, uniformly stirring the carbomer and the ultrapure water, fully swelling the carbomer overnight, adding triethanolamine to obtain blank hydrogel, adding the deer skin collagen polypeptide liposome into the blank hydrogel according to the proportion, and uniformly stirring the mixture to obtain the collagen polypeptide liposome hydrogel (DCP-LIP-Gel).

As shown in fig. 3, 5 and 6, fig. 3 is a graph showing a sustained release profile of an embodiment of a method for preparing a collagen polypeptide liposome hydrogel of deer skin according to the present application; fig. 5 is a graph of the results of skin permeation experiments according to one embodiment of the method for preparing a deer skin collagen polypeptide liposome hydrogel according to the present application, wherein n =3, P <0.001 compared to DCP group; fig. 6 is a graph showing the result of content stability of one example of the preparation method of the deer skin collagen polypeptide liposome hydrogel according to the present application, particularly, the result of content stability of different formulations of DCP at 4 ℃ (n = 3). Wherein the DCP is deer skin collagen polypeptide, the DCP-LIP is deer skin collagen polypeptide liposome, and the DCP-LIP-Gel is deer skin collagen polypeptide liposome hydrogel.

Furthermore, as shown in FIG. 7, FIG. 8 and FIG. 9, FIG. 7 is a graph showing the results of the change of the deer skin collagen polypeptide liposome hydrogel according to the present application to the skin of UVB-irradiated mice (N = 6), specifically, DCP-LIP and DCP-LIP-Gel to the skin of UVB-irradiated mice (1w, 3w,6w,9w in FIG. 7 mean 1 week, 3 weeks, 6 weeks, 9 weeks, N represents the normal control group, M represents the model group, and it can be seen from FIG. 7 that the change of the animal skin injury of the DCP-LIP-Gel group is minimal and the degree of the occurrence of no erythema and wrinkles is minimal, indicating that the protective effect of DCP-LIP-Gel on the skin photoaging of mice is strongest; FIG. 8 is a graph showing the results of physical stability of a deer skin collagen polypeptide liposome hydrogel according to the present application, specifically, appearance changes of DCP-LIP-Gel at 4 ℃ at different time points during storage, where 0m,1m,2m,3m,4m,5m in FIG. 8 represent 1 month, 2 months, 3 months, 4 months, 5 months, respectively; fig. 9 is a graph showing the results of the thermal-resistant, cold-resistant, and centrifugal stability of the deer skin collagen polypeptide liposome hydrogel according to the present application, specifically, the results of the thermal-resistant, cold-resistant, and centrifugal stability of DCP-LIP-Gel (deer skin collagen polypeptide liposome hydrogel), and it can be seen that the deer skin collagen polypeptide liposome hydrogel of the present application shows better stability at room temperature, 40 degrees celsius, -15 degrees celsius, and after centrifugation.

3. An application of the deer skin collagen polypeptide liposome hydrogel in improving skin cosmetic components is provided.

The deer skin collagen polypeptide liposome hydrogel improves the skin absorption and can realize the anti-aging effect. In particular, liposomes are the most suitable carriers for polypeptide delivery because they can satisfy the polar, non-polar and amphiphilic properties of protein hydrolysates. In addition, liposomes can be readily synthesized from food ingredients; they can capture and emit hydrophilic and hydrophobic elements and facilitate targeting. Nanocarriers improve the ability to cross biological barriers that otherwise are difficult for peptides to cross; in addition, they have a high surface area to volume ratio, which provides better solubility.

Bioactive polypeptides of smaller molecular weight can stimulate new cell synthesis of collagen and collagen fibers to improve skin wrinkles, fine lines and tone, thereby affecting different functions of the skin. The deer skin collagen polypeptide has good hydrophilicity, can be fully absorbed and utilized by skin to synthesize collagen, and has certain cosmetic effects (whitening, nourishing, moisturizing and anti-aging).

At present, many cosmetic compositions contain components such as collagen and collagen peptide to help maintain skin quality. Most are derived from animal skin or marine organisms. The hydrolyzed collagen on the market can be taken orally or externally (lotion, mask and moisturizing cream) to play the roles of moisturizing and resisting aging. The carrier system for the biologically active peptides is as important as the polypeptide itself. Carrier systems are effective methods for protecting biologically active peptides from degradation and for enhancing their activity, and they are also essential for the integration of biologically active peptides into commercial products. Many carrier systems have been developed including microemulsions, nanoemulsions, emulsions, solid lipid nanoparticles, liposomes, biopolymer nanoparticles, and hydrogel microbeads. Each method has advantages and limitations. Liposomes are the most suitable carriers for polypeptide delivery because they can satisfy the polar, non-polar and amphiphilic properties of protein hydrolysates. In addition, liposomes can be readily synthesized from food ingredients; they can capture and emit hydrophilic and hydrophobic elements and facilitate targeting. Nanocarriers improve the ability to cross biological barriers that otherwise are difficult for peptides to cross; in addition, they have a high surface area to volume ratio, which provides better solubility. In addition, a good carrier system should provide controlled release and enhanced bioavailability while minimizing side effects and toxicity. Liposomes can protect the bioactive substance from digestion in the stomach and promote absorption, thereby improving bioactivity and bioavailability. Meanwhile, the liposome has good biocompatibility, slow release property and targeting property. Therefore, it is entirely suitable to encapsulate the protein hydrolysate in liposomes.

Liposomes have been used as systemic drug delivery systems for many years because their biomimetic nature enables them to safely deliver water soluble and hydrophobic drugs over a relatively long period of time. Also, the biocompatibility of gels and their ability to retain certain types of drugs has led to the development of materials capable of sustained topical administration. Although both systems have been successful, there are drawbacks to using both systems alone. For example, if liposomes are cleared from the circulatory system within a few hours, the ability of certain liposome formulations to release drug within a few days becomes irrelevant. On the other hand, crosslinked gels have been demonstrated to be able to remain in vivo for up to a month or more; however, drug retention is generally dependent on whether or not a particular interaction exists between the drug and the gel. Thus, polypeptide drugs, such as fibroblast growth factor, may form polyionic complexes with the gel, resulting in long-term retention in the gel matrix, but low molecular compounds tend to diffuse out of the gel within hours. Therefore, the liposome and the gel are combined, so that a better substance suitable for long-term administration, namely the deer skin collagen polypeptide liposome hydrogel, can be obtained.

The application has the following beneficial effects:

1. novel vector systems are provided: the carrier system for biologically active peptides is as important as the polypeptide itself. Carrier systems are effective methods for protecting biologically active peptides from degradation and for enhancing their activity, and they are also essential for the integration of biologically active peptides into commercial products. Liposomes are provided as novel carrier systems.

2. Improved contact time with the skin: the use of liposomes as carriers can improve penetration of drugs through the skin by controlling drug release, increasing the residence time of the drug on the skin, allowing direct contact with the stratum corneum or skin appendages, and reducing the physicochemical instability (osmotic) activity of the drug.

3. The storage stability is improved: the deer skin collagen polypeptide is wrapped in the liposome to prepare hydrogel, so that the physical stability, the chemical stability and the like of the deer skin collagen polypeptide are improved.

4. And (3) promoting skin penetration: the deer skin collagen polypeptide has an undesirable skin permeation effect, and the deer skin collagen polypeptide is prepared into a liposome form, namely the deer skin collagen polypeptide liposome hydrogel, so that the skin permeation is effectively promoted.

5. The deer skin collagen polypeptide liposome hydrogel can play a better role in skin photoaging, and has more obvious effect than that of liposome and free drugs.

It should be understood that, although the steps in the flowcharts of the figures are shown in order as indicated by the arrows, the steps are not necessarily performed in order as indicated by the arrows. The steps are not performed in the exact order shown and may be performed in other orders unless explicitly stated herein. Moreover, at least a portion of the steps in the flow chart of the figure may include multiple sub-steps or multiple stages, which are not necessarily performed at the same time, but may be performed at different times, which are not necessarily performed in sequence, but may be performed alternately or alternately with other steps or at least a portion of the sub-steps or stages of other steps.

It should be understood that the above-described embodiments are merely exemplary of some, and not all, embodiments of the present application, and that the drawings illustrate preferred embodiments of the present application without limiting the scope of the claims appended hereto. This application is capable of embodiments in many different forms and the embodiments are provided so that this disclosure will be thorough and complete. Although the present application has been described in detail with reference to the foregoing embodiments, it will be apparent to one skilled in the art that modifications can be made to the embodiments described in the foregoing detailed description, or equivalents can be substituted for some of the features described therein. All equivalent structures made by using the contents of the specification and the drawings of the present application are directly or indirectly applied to other related technical fields and are within the protection scope of the present application.

Claims (11)

1. The deer skin collagen polypeptide liposome hydrogel is characterized in that the deer skin collagen polypeptide liposome hydrogel is prepared by every 100ml of solution, and comprises the following components:

10-50 g of deer skin collagen polypeptide, 20-100 g of egg yolk lecithin, 10-20 g of cholesterol, 2-5 g of tween-80, 2-10 g of triethanolamine, 2-10 g of carbomer and 100ml of chloroform.

2. The deer skin collagen polypeptide liposome hydrogel according to claim 1, wherein said deer skin collagen polypeptide liposome hydrogel is prepared from per 100ml solution, comprising:

10g of deer skin collagen polypeptide, 20g of egg yolk lecithin, 10g of cholesterol, 2g of tween-80, 2g of triethanolamine, 2-10 g of carbomer and 100ml of chloroform.

3. A preparation method of deer skin collagen polypeptide liposome hydrogel is characterized by comprising the following steps:

preparing deer skin collagen polypeptide;

dissolving egg yolk lecithin, cholesterol and tween-80 in a chloroform solvent to obtain a mixed solution;

spin-drying the mixed solution to be transparent film-shaped to obtain liposome, adding a solution containing the deer skin collagen polypeptide into the liposome, and hydrating to obtain deer skin collagen polypeptide liposome;

placing the deer skin collagen polypeptide liposome in a centrifuge tube, oscillating the centrifuge tube, performing ultrasonic treatment with an ice-water bath probe to obtain the deer skin collagen polypeptide liposome which is uniformly dispersed;

uniformly mixing carbomer weighed in advance with ultrapure water, standing overnight, and adding triethanolamine to obtain blank hydrogel;

and adding the deer skin collagen polypeptide liposome into the blank hydrogel to obtain the collagen polypeptide liposome hydrogel.

4. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 3, wherein the step of preparing deer skin collagen polypeptide comprises:

preparing deer skin collagen;

adding alkaline protease into the deer skin collagen for hydrolysis reaction, and inactivating enzyme in boiling water after 4h to obtain hydrolysate;

and cooling and centrifuging the hydrolysate, taking supernatant, and freeze-drying to obtain the dried deer skin collagen polypeptide.

5. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 4, wherein the step of preparing deer skin collagen comprises:

dissolving deer skin glue powder in acetic acid water solution, magnetically stirring, and filtering to obtain filtrate;

centrifuging the filtrate, and taking a supernatant, wherein the supernatant is a crude acid-soluble collagen solution;

salting out the acid-soluble crude collagen solution to obtain a crude collagen product;

dissolving the collagen crude product in acetic acid solution, dialyzing in 0.1M acetic acid water solution for 48 hr, dialyzing in ultrapure water for 24 hr, and freeze-drying to obtain the deer skin collagen.

6. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 5, wherein said step of salting out said acid-soluble collagen crude solution to obtain a collagen crude product comprises:

adding sodium chloride into the acid-soluble collagen crude solution until the final concentration of the sodium chloride is 2.0mol/L, centrifuging, and retaining the precipitate to obtain the collagen crude product.

7. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 5, wherein the concentration of said aqueous acetic acid solution is 0.5mol/L.

8. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 5, wherein the mass ratio of the deer skin gelatin powder to the acetic acid aqueous solution is 1.

9. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 3, wherein the molar ratio of egg yolk lecithin to cholesterol is 3.

10. The method for preparing the deer skin collagen polypeptide liposome hydrogel according to claim 3, wherein the step of spin-drying the mixed solution into a transparent film shape comprises:

and slowly vacuumizing the mixed solution at the rotation speed of 90rmp, carrying out water bath at the temperature of 30 ℃, and spin-drying the mixed solution to form a transparent film.

11. Use of the deer skin collagen polypeptide liposome hydrogel of claim 1 for improving skin cosmetic components.

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