A collagen drink containing loaded PDRN vesicle spheres and a method of preparation thereof

Abstract

The application discloses a collagen drink containing PDRN vesicle balls and a preparation method thereof, and belongs to the technical field of biotechnology. The preparation method of the collagen drink containing PDRN vesicle balls comprises the following steps: mixing PDRN vesicle balls and polysaccharides, and carrying out gelation reaction and homogenization to obtain a suspension containing the PDRN vesicle balls. The application provides a method for successfully preparing the collagen drink containing PDRN vesicle balls. The PDRN vesicle balls can stably exist in the preparation process of the drink and maintain original biological activity. The collagen drink containing PDRN vesicle balls prepared by the method has a wrinkle area reduction rate of nearly 20% and a collagen density increase rate of more than 50% after being taken for 28 days.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to a collagen drink containing PDRN-loaded vesicle spheres and a preparation method thereof. BACKGROUND

[0002] Polydeoxyribonucleotide (PDRN) is a DNA derivative with a molecular weight ranging from 50 to 1500 kDa, which has various pharmacological activities such as promoting tissue repair, wound healing and stimulating angiogenesis. It has been confirmed that PDRN can participate in the repair of intracellular DNA. It can promote the biosynthesis of mitochondria by activating the hope factor (PGC-1a) in mitochondria through the cAMP signaling pathway, repair cell damage, restore cell health, and accelerate the synthesis of collagen. However, PDRN is prone to inactivation and failure, and its large molecular weight makes it difficult to be absorbed and utilized by the human body, resulting in the difficulty of PDRN in food products to achieve popularization and application.

[0003] In recent years, the collagen drink market has shown a rapid growth trend, especially the consumption group of collagen drink for preventing skin aging is expanding. Skin aging is caused by cell damage and collagen loss due to exogenous and endogenous factors. Exogenous factors include sun exposure, seasonal change and environmental pollution, and endogenous factors include age, sleep deprivation and stress. The key to anti-aging from the root is cell damage repair and collagen supplementation.

[0004] In previous studies, it was found that the activity and stability of PDRN in human digestive juice can be improved by loading it in a vesicle structure. Vesicles are liposome substances secreted by cells, which have a unique structure that can be used as a delivery system for bioactive ingredients. According to the source, it can be divided into human, animal, plant and microbial sources. The separation and extraction methods, especially the separation and extraction methods of animal and plant vesicles, mainly include two types. One is to crush animal and plant tissues to extract cells or vesicle fragments, and then recombine to obtain vesicles. The other method is mainly based on tissue penetration to collect exosomes to extract vesicles in a way that preserves the integrity of the vesicles. For example, patent application 202510324022.1 discloses a method of loading PDRN on animal and plant vesicle spheres obtained by crushing and recombination, which can effectively solve the problems of PDRN degradation in the gastrointestinal tract and difficulty in membrane absorption. However, there is no report on the application of PDRN-loaded vesicle spheres to drinks. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a collagen drink containing loaded PDRN vesicle balls and a preparation method thereof, which successfully provides a method for preparing the collagen drink containing loaded PDRN vesicle balls, and the loaded PDRN vesicle balls can stably exist in the preparation process of the drink and maintain the original biological activity.

[0006] Compared with the prior art, the present application has the following advantages:

[0007] 1. The preparation method of the collagen drink containing loaded PDRN vesicle balls comprises gelation reaction and homogenization of polysaccharides and loaded PDRN vesicle balls, and then compounding with collagen to prepare the collagen drink containing loaded PDRN vesicle balls. The loaded PDRN vesicle balls can stably exist in the preparation process of the drink and maintain the original biological activity. The collagen drink containing loaded PDRN vesicle balls has a significant effect of reducing wrinkles and increasing facial collagen density.

[0008] 2. The method for preparing the collagen drink containing loaded PDRN vesicle balls uses the intermolecular interaction between polysaccharides and loaded PDRN vesicle balls and hydration to construct a protective layer on the surface of the loaded PDRN vesicle balls and improve the mechanical strength of the membrane structure of the loaded PDRN vesicle balls, without cross-linking, thereby avoiding the influence of non-edible reagents such as additional cross-linking agents on food products.

[0009] 3. Preferably, the method for preparing the collagen drink containing loaded PDRN vesicle balls comprises homogenizing the system using carbon dioxide pressure medium after gelation, which can effectively avoid the influence of polar medium on the system and realize the generation of hydration layer and the wrapping of loaded PDRN vesicle balls.

[0010] The technical solutions of the present application will be further described in detail below in combination with the drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 SEM image of loaded PDRN vesicle balls in the collagen drink containing loaded PDRN vesicle balls of Example 1;

[0012] Figure 2 SEM image of loaded PDRN vesicle balls in the collagen drink containing loaded PDRN vesicle balls of Comparative Example 1;

[0013] Figure 3 SEM image of loaded PDRN vesicle balls in the collagen drink containing loaded PDRN vesicle balls of Comparative Example 2;

[0014] Figure 4 SEM image of PDRN-loaded vesicle spheres in collagen drink for Example 2 containing PDRN-loaded vesicle spheres;

[0015] Figure 5 SEM image of PDRN-loaded vesicle spheres in collagen drink for Comparative Example 3 containing PDRN-loaded vesicle spheres and partial magnification;

[0016] Figure 6 SEM image of PDRN-loaded vesicle spheres in collagen drink for Comparative Example 4 containing PDRN-loaded vesicle spheres;

[0017] Figure 7 Schematic diagram of changes in facial collagen of volunteers using collagen drink containing PDRN-loaded vesicle spheres for Example 2, Comparative Example 3 or Comparative Example 4;

[0018] Figure 8 SEM image of PDRN-loaded vesicle spheres in collagen drink for Example 3 containing PDRN-loaded vesicle spheres;

[0019] Figure 9 SEM image of PDRN-loaded vesicle spheres in collagen drink for Comparative Example 5 containing PDRN-loaded vesicle spheres;

[0020] Figure 10 Schematic diagram of changes in facial collagen of volunteers using collagen drink containing PDRN-loaded vesicle spheres for Example 3 or Comparative Example 5. DETAILED DESCRIPTION

[0021] The technical solutions will be described clearly and completely below in conjunction with the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0022] In the following description, the term "and / or" is used to describe the association relationship of the associated objects, which means that there can be three relationships, for example, A and / or B, which can represent the cases of A alone, B alone and A and B together. Wherein A and B can be singular or plural.

[0023] In the following description, the terms "include", "contain", "have" and "contain" and the like are all open terms, that is, they mean to include but not limited to.

[0024] Those skilled in the art shall understand that in the following description of the embodiments of the present application, the order of the serial numbers does not mean the order of execution, and part or all of the steps can be executed in parallel or in sequence, and the execution order of each process shall be determined according to its function and inherent logic, and shall not constitute any limitation on the implementation process of the embodiments of the present application.

[0025] Those skilled in the art shall understand that the numerical ranges in the embodiments of the present application shall be understood as also specifically disclosing each intermediate value between the upper limit and the lower limit of the range. Each smaller range between any stated value or stated range of values and any other stated value or stated range of values is also included within the present application. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0026] Unless otherwise specified, the technical / scientific terms used herein have the same meaning as generally understood by those skilled in the art to which the present application belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present application. All documents mentioned in the specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In the event of conflict between the content of the specification and any incorporated document, the content of the specification shall prevail.

[0027] The technical principle adopted by the present application is to use polysaccharides to form a gel structure on the surface of the vesicle ball as a barrier to prevent external enzymes or oxidants from damaging the PDRN-loaded vesicle ball, to avoid deformation or rupture of the PDRN-loaded vesicle ball due to excessive water absorption or dehydration, to resist the impact of mechanical stress on the PDRN-loaded vesicle ball, and to combine the polysaccharides with the membrane lipid components of the PDRN-loaded vesicle ball to stabilize the membrane structure of the vesicle.

[0028] In one aspect, a method for preparing a collagen drink containing PDRN-loaded vesicle balls is provided, comprising: mixing PDRN-loaded vesicle balls and polysaccharides, and performing a gelation reaction and homogenization to obtain a suspension containing PDRN-loaded vesicle balls.

[0029] The method for preparing a collagen drink containing PDRN-loaded vesicle balls of the present application comprises mixing PDRN-loaded vesicle balls and polysaccharides, and performing a gelation reaction and homogenization to obtain a suspension containing PDRN-loaded vesicle balls. The gelation and homogenization are used to realize the interaction between the polysaccharides and the PDRN-loaded vesicle balls, to form a gel barrier containing a hydration layer on the surface of the PDRN-loaded vesicle balls, to combine the polysaccharide-loaded PDRN vesicle ball membrane components through intermolecular forces, to improve the membrane structure, and to enhance the mechanical strength of the PDRN-loaded vesicle ball membrane. The collagen drink containing PDRN-loaded vesicle balls prepared by the method of the present application has a significantly enhanced effect of reducing wrinkles and enhancing facial collagen density.

[0030] In some embodiments, the PDRN-loaded vesicle spheres are PDRN-loaded vesicle spheres obtained by crushing and recombining cells or cell secretions of animal or plant origin with PDRN. In some preferred embodiments, the vesicle spheres are derived from plant sources. In some preferred embodiments, the PDRN-loaded vesicle spheres have a size of 80-100 nm.

[0031] Preferably, the PDRN-loaded vesicle spheres of the present application are PDRN-loaded vesicle spheres obtained by crushing and recombining cells or cell secretions of animal or plant origin with PDRN. The crushing and recombination is based on crushing tissues of animal or plant origin to obtain cell fragments and vesicle fragments, and recombining the active ingredient PDRN to obtain PDRN-loaded vesicle spheres. During the research, the inventors found that such vesicle spheres obtained by crushing and recombination have a structure similar to that of a vesicle membrane, and can effectively resist damage from substances such as pepsin, trypsin, and pancreatic amylase in digestive fluids such as gastric juice and intestinal juice. The vesicle spheres can stably exist in the environment of gastric acid and intestinal contents. However, the collagen drink prepared using the PDRN-loaded vesicle spheres as raw materials needs to undergo raw material pretreatment, blending, and sterilization of the blended material. The preparation process is long, the environmental conditions change rapidly, and the process involves mechanical pressure such as stirring, which can damage the PDRN-loaded vesicle spheres, causing them to lose water and deform, and thus the collagen drink containing the PDRN-loaded vesicle spheres cannot maintain the inherent biological activity of the PDRN-loaded vesicle spheres. By performing a gelation reaction and homogenization of the polysaccharides and the PDRN-loaded vesicle spheres before blending with the collagen components, a hydration barrier can be constructed on the surface of the recombined PDRN-loaded vesicle spheres. The polysaccharides are bonded to the phospholipids in the membrane structure of the recombined PDRN-loaded vesicle spheres, stabilizing the membrane structure and improving the mechanical strength of the lipid membrane on the surface of the PDRN-loaded vesicle spheres. In some preferred embodiments, the PDRN-loaded vesicle spheres are PDRN-loaded vesicle spheres obtained by crushing and recombining cells or cell secretions of plant origin. Each vesicle sphere contains more than 80,000 PDRN molecules. The collagen drink containing the PDRN-loaded vesicle spheres obtained by the method of the present application has a complete structure, and can achieve collagen supplementation, collagen repair, and collagen regeneration, and promote the recovery of biological cell health in the whole link, and has the effect of accelerating collagen synthesis.

[0032] In some specific embodiments, the above method comprises:

[0033] Mixing the PDRN-loaded vesicle spheres, the polysaccharides, and pure water, and heating and stirring to gelate, to obtain a gel liquid;

[0034] Homogenizing the gel liquid to obtain a gel suspension;

[0035] Mixing the gel suspension with collagen, and filling and sterilizing to obtain a collagen drink containing PDRN-loaded vesicle spheres.

[0036] The present application does not limit the source of pure water, which meets the requirements of the beverage standards. The homogenization, mixing equipment, filling and sterilization device are not limited, and the commonly used equipment in the field can be used.

[0037] In some embodiments, the mass ratio of PDRN to polysaccharide in the loaded PDRN vesicle sphere is (1:2) to (2:1). Preferably, the mass ratio of PDRN to polysaccharide is 1:1.

[0038] Preferably, the present application improves the stability and mechanical strength of the loaded PDRN vesicle sphere by gelation and homogenization of polysaccharide and loaded PDRN vesicle sphere. When the mass ratio of PDRN to polysaccharide in the loaded PDRN vesicle sphere is 1:2 to 2:1, the polysaccharide can form a dense hydrophilic layer on the surface of the loaded PDRN vesicle sphere, interact with the loaded PDRN vesicle sphere through intermolecular interaction, protect the loaded PDRN vesicle sphere from external factors, reduce the van der Waals force between the loaded PDRN vesicle spheres through steric hindrance effect, effectively avoid the aggregation or fusion of the loaded PDRN vesicle spheres, increase the mechanical strength of the membrane structure, and resist the impact of mechanical stress. Especially when the mass ratio of PDRN to polysaccharide is 1:1, the hydration layer on the surface of the vesicle sphere in the beverage is the most dense, and the obtained beverage has a significant effect of reducing the surface wrinkle area. PDRN is commercially available, and the source of PDRN is not limited in the present application. In the following examples, PDRN with a purity of 80% purchased from Shanghai Huivon Biotechnology Co., Ltd. is used as an example.

[0039] In some embodiments, the polysaccharide is sodium hyaluronate.

[0040] Preferably, in the process of constructing stable loaded PDRN vesicle spheres by gelation and homogenization, when the polysaccharide is sodium hyaluronate, the hydroxyl and carboxyl groups of sodium hyaluronate can fully combine with water molecules through hydrogen bonds, forming a hydration layer on the surface of the loaded PDRN vesicle sphere, and avoiding the imbalance of internal osmotic pressure of the loaded PDRN vesicle sphere.

[0041] In some embodiments, the heating temperature for gelation is 65-75℃, and the stirring rate is 500-800 rpm. In some preferred embodiments, the heating temperature is 75℃, and the stirring rate is 500-650 rpm.

[0042] In some specific embodiments, the temperature for gelation is 65-75℃, and the stirring rate is 500-800 rpm, which can realize the formation of the hydration layer on the surface of the loaded PDRN vesicle sphere and the interaction between the polysaccharide and the membrane components of the loaded PDRN vesicle sphere. Preferably, the temperature is 75℃, and the stirring rate is 500-650 rpm, the hydration layer structure is the most dense, the structure of the loaded PDRN vesicle sphere in the obtained beverage is the most complete, and the beverage has a significant effect of reducing facial wrinkles and increasing facial collagen density.

[0043] In some embodiments, the homogenization pressure is 12-18 MPa. In some preferred embodiments, the homogenization pressure is 13-15 MPa.

[0044] In some embodiments, the homogenization pressure medium is carbon dioxide.

[0045] In some specific embodiments, the homogenization pressure is 12-18 MPa, and the homogenization pressure medium is CO2, which can sufficiently realize the generation of hydration layer and the wrapping of the PDRN-loaded vesicle sphere. When the CO2 pressure is too low, the homogenization cannot be realized sufficiently, and when the CO2 pressure is too high, the PDRN-loaded vesicle sphere in the beverage is irregular in shape, the surface stress is uneven, and even ruptures. Preferably, the homogenization pressure is 13-15 MPa, and the PDRN-loaded vesicle sphere obtained in the beverage is clear in boundary, high in shape regularity, and the PDRN is distributed in the center of the vesicle sphere, and the beverage has significant wrinkle-reducing, anti-aging and facial collagen density-promoting effects.

[0046] In some embodiments, the mass of the pure water is 500 times the mass of the PDRN.

[0047] In some embodiments, the collagen protein includes collagen peptide and elastin.

[0048] In some embodiments, the mass of the collagen protein in the collagen protein beverage containing the PDRN-loaded vesicle sphere is 120-125 times the mass of the gel suspension.

[0049] In some embodiments, the mass percentage of the collagen protein in the collagen protein beverage containing the PDRN-loaded vesicle sphere is 24-25%.

[0050] In another aspect, a beverage prepared by the method for preparing the collagen protein beverage containing the PDRN-loaded vesicle sphere is provided.

[0051] The present application has undergone a series of experiments before the application, and now some test results are listed to further describe the application in detail, which is described in detail below in combination with the embodiments.

[0052] Embodiment 1

[0053] The present embodiment provides a method for preparing a collagen protein beverage containing a PDRN-loaded vesicle sphere, which comprises:

[0054] Step one, mix the loaded PDRN vesicle ball and 50 mg of sodium hyaluronate, add purified water, make up to 50 g, heat the system to 75℃ after constant volume and keep, stir at 500 rpm for 10 min, get the loaded PDRN vesicle ball gel liquid; the weight average molecular weight of the sodium hyaluronate is 81K-100KDa, the viscosity value is 2.85dl / g; the mass of PDRN loaded in the loaded PDRN vesicle ball is 50 mg; the preparation method of the loaded PDRN vesicle ball refers to the patent application file CN202510324022.1 A food-derived biological vesicle loaded with active ingredients and its preparation method and application, specifically the method recorded in Example 2, and the loaded PDRN vesicle ball is the food-derived biological vesicle loaded with polydeoxyribonucleotides prepared in Example 2;

[0055] Step two, transport the loaded PDRN vesicle ball gel liquid into a homogenizing device, pass in carbon dioxide to make the pressure reach 13 MPa and keep, homogenize for 0.2 h, get the loaded PDRN vesicle ball gel suspension;

[0056] Step three, in a dispensing tank, according to the total amount of 100 g, mix 200 mg of loaded PDRN vesicle ball gel suspension, 24 g of fish collagen peptide, 200 mg of katsuwonus pelamis elastin, 200 mg of aurantias hortensis concentrated powder, 10 g of lime lemon concentrated juice and 0.2 g of citric acid, and the rest is made up with purified water, stir at 500 rpm to mix evenly, complete the dispensing; the molecular weight of fish collagen peptide is 500 Da, purchased from Hubei Jianpeibio Technology Co., Ltd.; Katsuwonus pelamis elastin is purchased from Zhengyi Health Group; The concentration of (L-ergothioneine) in Aurantias hortensis concentrated powder is ≥1.0%, purchased from Shanghai Ergothioneine Biotechnology Group Co., Ltd.; The sugar content of lime lemon concentrated juice is 55.0±1.0, purchased from Green Spring Biotechnology Co., Ltd.; Citric acid is anhydrous citric acid, purchased from Zhengzhou Jiuxing Biological Technology Co., Ltd.;

[0057] Step four, after dispensing, sterilize at 105℃ for 25 min to get the collagen drink containing loaded PDRN vesicle ball.

[0058] Comparative example 1

[0059] This comparative example examines the influence of polysaccharide type on the performance of collagen drink containing loaded PDRN vesicle ball, the preparation method is the same as example 1, the only difference is that in step one, an equal amount of β-glucan is used to replace sodium hyaluronate; the β-glucan is derived from yeast, and the weight average molecular weight is 1 million.

[0060] Comparative example 2

[0061] The comparative example examines the influence of the amount of polysaccharide added on the performance of the collagen drink containing PDRN-loaded vesicle spheres. The preparation method is the same as that of Example 1, with the only difference being that in Step 1, the mass of sodium hyaluronate is 100 mg.

[0062] The structure of the PDRN-loaded vesicle spheres in the collagen drink containing PDRN-loaded vesicle spheres of Example 1, the structure of the PDRN-loaded vesicle spheres in the collagen drink containing PDRN-loaded vesicle spheres of Comparative Example 1, and the structure of the PDRN-loaded vesicle spheres in the collagen drink containing PDRN-loaded vesicle spheres of Comparative Example 2 were characterized by scanning electron microscopy, and the results are shown in FIGS. 1-3, respectively. Figures 1-3 Figure 1 FIG. 1 is a SEM image of the structure of the PDRN-loaded vesicle spheres in the collagen drink containing PDRN-loaded vesicle spheres of Example 1. Figure 2 FIG. 2 is a SEM image of the structure of the PDRN-loaded vesicle spheres in the collagen drink containing PDRN-loaded vesicle spheres of Comparative Example 1. Figure 3 FIG. 3 is a SEM image of the structure of the PDRN-loaded vesicle spheres in the collagen drink containing PDRN-loaded vesicle spheres of Comparative Example 2. Figures 1-3 As can be seen, in the collagen drink containing PDRN-loaded vesicle spheres of Example 1, the PDRN-loaded vesicle spheres are uniform and transparent, with clear edges and complete structure, and the PDRN located at the center of the vesicle spheres can be clearly observed. In the collagen drink containing PDRN-loaded vesicle spheres of Comparative Example 1, the vesicle spheres are severely damaged, and obvious separation of the vesicle lipid layer can be observed. In the collagen drink containing PDRN-loaded vesicle spheres of Comparative Example 2, the transparency of the vesicle spheres is reduced.

[0063] Volunteers were selected: age 30-45 years; skin wrinkles were obvious, and the crow's feet were 2-4 levels (according to the Rao-Goldman facial wrinkle grading standard); the effect was determined by before and after comparison; the test site was the face, the room temperature was 21.0°C±1.0°C, and the humidity was 50%±10% during the test; the skin collagen density was tested by Ultrascan UC22, and the area ratio of the crow's feet (fish tail wrinkle) wrinkle was tested by Visia; the follow-up cycle was set to 0, 7, 17, and 28 days. The volunteers in the experimental group took one bottle of 50 mL of the collagen drink containing PDRN-loaded vesicle spheres per day.

[0064] ​The changes of wrinkles on the eyes of the volunteers taking the collagen beverage containing the PDRN vesicle ball of the embodiment 1 or the comparative examples 1-2 were recorded, and the results are shown in Table 1. The change rate of the wrinkle area was calculated as (initial wrinkle area - wrinkle area after use for a predetermined number of days) / initial wrinkle area. According to Table 1, after use for 28 days, the experimental groups all showed different degrees of decrease in the wrinkle area, the blank control group showed an increase in the wrinkle area, and the increase rate was 3.61%. The decrease rate of the wrinkle area of the embodiment 1 group was 18.21%, the decrease rate of the wrinkle area of the comparative example 1 group was 9.94%, and the decrease rate of the wrinkle area of the comparative example 2 group was 11.43%. The decrease rate of the wrinkle area of the embodiment 1 group was obviously higher than those of the comparative example groups and the control group, indicating that the collagen beverage containing the PDRN vesicle ball of the application can effectively promote the repair of damaged cells and restore the young state of the skin.

[0065] Table 1 Changes of wrinkles on the eyes of volunteers after taking the products of the embodiment 1, the comparative example 1 and the comparative example 2

[0066]

[0067] Embodiment 2

[0068] The embodiment is the same as the embodiment 1, except that in step one, the stirring speed is 650 rpm.

[0069] Comparative Example 3

[0070] The comparative example examines the influence of the gelation reaction temperature on the performance of the collagen beverage containing the PDRN vesicle ball. The preparation method is the same as that of the embodiment 2, except that in step one, the temperature for heating and maintaining the constant volume system is 70℃.

[0071] Comparative Example 4

[0072] The comparative example examines the influence of the gelation reaction condition on the performance of the collagen beverage containing the PDRN vesicle ball. The preparation method is the same as that of the embodiment 2, except that in step one, the stirring speed is 800 rpm.

[0073] The structure scanning electron microscope results of the PDRN vesicle ball in the collagen beverage containing the PDRN vesicle ball of the embodiment 2 and the comparative examples 3-4 are shown in Table 2. Figures 4-6 Table 2 Structure scanning electron microscope results of the PDRN vesicle ball in the collagen beverage containing the PDRN vesicle ball of the embodiment 2 and the comparative examples 3-4 Figure 4 is the SEM image of the structure of the PDRN vesicle ball in the collagen beverage containing the PDRN vesicle ball of the embodiment 2, Figure 5 is the SEM image and the local enlarged image of the structure of the PDRN vesicle ball in the collagen beverage containing the PDRN vesicle ball of the comparative example 3, Figure 6 is the SEM image of the structure of the PDRN vesicle ball in the collagen beverage containing the PDRN vesicle ball of the comparative example 4. Figures 4-6As can be seen, the PDRN-loaded vesicles in Example 2 are uniform, transparent, and structurally intact, with PDRN clearly visible at the center of the vesicles. In Comparative Example 3, floating polysaccharide components are clearly visible in the scanning electron microscope field, failing to form a complete gel structure. The PDRN-loaded vesicles in Comparative Example 4 show obvious rupture.

[0074] Changes in collagen levels were recorded in volunteers who consumed collagen drinks containing PDRN-loaded vesicles as described in Example 2 or Comparative Examples 3-4. The results are as follows: Figure 7 As shown in Table 2, after 28 days of use, the facial collagen of volunteers in both Example 1 and Comparative Examples 1 and 2 increased to varying degrees. The quantified results of collagen changes are shown in Table 2. The rate of change in facial collagen density was calculated as (initial facial collagen density - facial collagen density after the preset number of days of use) / initial facial collagen density. According to Table 1, after 28 days of use, all groups showed varying degrees of increase in collagen density. The facial collagen density of the blank control group increased by 16.77%, the Example 2 group increased by 50.14%, the Comparative Example 3 group increased by 30.18%, and the Comparative Example 4 group increased by 28.35%. Compared with the blank control group, the collagen drink containing PDRN-loaded vesicles of Example 2 of this invention has a significant effect on promoting increased facial collagen density.

[0075] Table 2. Changes in facial collagen density after volunteers took the products in Examples 2 and Comparative Examples 3-4.

[0076]

[0077]

[0078] Example 3

[0079] This embodiment is the same as Embodiment 1, except that in step two, the homogenization pressure is 15 MPa.

[0080] Comparative Example 5

[0081] This comparative study investigated the effect of homogenization pressure on the performance of collagen drinks containing PDRN-loaded vesicles. The preparation method was the same as in Example 3, except that the homogenization pressure was 18 MPa.

[0082] Scanning electron micrographs of the PDRN-loaded vesicle structures in collagen drinks containing PDRN-loaded vesicles in Example 3 and Comparative Example 5 are shown below. Figures 8-9 As shown, where Figure 8 This is a SEM image of the PDRN-loaded vesicle structure in the collagen beverage containing PDRN-loaded vesicles in Example 3. Figure 9 The image shows the SEM image of the PDRN-loaded vesicle structure in the collagen drink containing PDRN-loaded vesicles in Comparative Example 5.Figures 8-9 It can be seen that the loaded PDRN vesicle spheres of Example 3 are clear in edge, complete in structure, and the PDRN located at the center of the vesicle spheres can be obviously observed. The loaded PDRN vesicle spheres of Comparative Example 5 are irregular in shape, and present a state of being about to break.

[0083] The changes of wrinkles and the changes of facial collagen density of the volunteers who took the collagen drinks containing the loaded PDRN vesicle spheres of Example 3 and Comparative Example 5 were tested according to the above method, and the results are shown in Table 3 and Table 4. Figure 10 , Table 3 and Table 4,

[0084] Table 3 Changes of wrinkles of the volunteers who took the products of Example 3 and Comparative Example 5 for 28 days

[0085]

[0086] Table 4 Changes of facial collagen density of the volunteers who took the products of Example 3 and Comparative Example 5 for 28 days

[0087]

[0088] According to Table 3, after taking for 28 days, the area of wrinkles of the blank control group is reduced by 2.72%, the area of wrinkles of Example 3 group is reduced by 19.83%, and the area of wrinkles of Comparative Example 5 group is reduced by 11.49%. The effect of removing wrinkles of Example 3 group is obviously higher than that of Comparative Example and the blank control group, which indicates that the collagen drink containing the loaded PDRN vesicle spheres prepared by the method of the present application has obvious wrinkle-removing performance.

[0089] According to Table 4, Figure 10 It can be seen that after taking for 28 days, the facial collagen density of the blank control group is increased by 10.79%, the facial collagen density of Example 3 group is increased by 37.15%, and the facial collagen density of Comparative Example 5 group is increased by 26.64%, which indicates that the collagen drink containing the loaded PDRN vesicle spheres prepared by the method of the present application has obvious effect of increasing facial collagen density.

Claims

1. A method of preparing a collagen drink containing PDRN-loaded vesicle spheres, characterized by, The application relates to a method for preparing a collagen drink containing PDRN vesicle balls. The PDRN vesicle balls, polysaccharides and pure water are mixed, heated and stirred to gelate, so as to obtain a gel liquid; The gel liquid is homogenized to obtain a gel suspension; The gel suspension is mixed with collagen, filled and sterilized to obtain the collagen drink containing the PDRN vesicle balls; The mass ratio of PDRN to the polysaccharides in the PDRN vesicle balls is 1:1; the heating temperature for gelation is 75 DEG C; the stirring speed for gelation is 500-650 rpm; and the pressure for homogenization is 13-15 MPa; The polysaccharides are sodium hyaluronate.

2. The method of claim 1, wherein the collagen drink containing PDRN-loaded vesicle spheres is prepared by, The PDRN vesicle balls are obtained by crushing and recombining animal and plant cell or cell secretion and PDRN.

3. The method of claim 1, wherein the collagen drink containing the PDRN-loaded vesicle sphere is prepared by, The pressure medium for homogenization is carbon dioxide.

4. The method of claim 1, wherein the collagen drink containing the PDRN-loaded vesicle sphere is prepared by, The collagen drink containing the PDRN vesicle balls comprises collagen peptides and elastin.

5. The method of claim 1, wherein the collagen drink containing PDRN-loaded vesicle spheres is prepared by, The mass of the collagen in the collagen drink containing the PDRN vesicle balls is 120-125 times the mass of the gel suspension; and / or the mass percentage of the collagen in the collagen drink containing the PDRN vesicle balls is 24-25%.

6. A collagen drink prepared by the method for preparing the collagen drink containing the PDRN vesicle balls according to claim 1.

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