Pdrn-metal ion microsphere emulsion with antioxidant and microecological balance effects, and preparation method and application thereof

Abstract

The application relates to the cosmetic technical field and discloses PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects and a preparation method and application thereof. The preparation method comprises the following steps: preparing a PDRN aqueous solution and a metal salt aqueous solution; and the metal salt aqueous solution is dropped into the PDRN aqueous solution and stirred to obtain the PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects. According to the microspherization technology, the PDRN is stably compounded with specific metal ions and integrated in an emulsion system, the endogenous antioxidant pathway is activated through the PDRN, and the metal ions directly remove free radicals to form synergy, so that the antioxidant efficiency is significantly improved; the mild bacteriostatic property of the metal ions is utilized to synergistically regulate the skin microecology, the oxidation damage caused by the imbalance of the flora is reduced from the source, and a double-dimensional anti-aging mechanism of "removing free radicals-regulating microecology" is constructed; not only are the problems of stability and transdermal absorption of active ingredients solved, but also long-term activity of the active ingredients in a complex emulsion matrix is ensured.

Description

Technical Field

[0001] This application relates to the field of cosmetic technology, specifically to a PDRN-metal ion microsphere emulsion with antioxidant and microecological balancing effects, its preparation method, and its application. Background Technology

[0002] With the accelerated pace of modern life and intensified environmental pollution, human skin is constantly exposed to external stressors such as ultraviolet radiation and air pollutants, leading to an excessive accumulation of reactive oxygen species (ROS) and triggering oxidative stress. This is the core mechanism behind photoaging phenomena such as collagen degradation, lipid peroxidation, cell damage, and even dull skin tone and wrinkle formation. Therefore, developing highly effective antioxidants is an important direction in the field of anti-aging cosmetics. Currently used antioxidant ingredients such as vitamin C, vitamin E, and polyphenols generally suffer from poor stability, low transdermal absorption efficiency, and limited and short-lasting effects, which restricts their full efficacy.

[0003] Meanwhile, the skin microbiome, as the outermost biological barrier of the human body, is crucial for maintaining skin health. Dysbiosis of the skin surface flora, particularly the excessive proliferation of potentially harmful bacteria, is not only associated with inflammatory problems such as acne and dermatitis, but recent research also indicates that the metabolic activity of certain harmful bacteria can exacerbate local oxidative stress and is linked to the skin aging process. Therefore, in anti-aging strategies, simply scavenging free radicals is no longer sufficient; synergistic regulation of the skin microbiome is becoming a new cutting-edge trend.

[0004] Polydeoxyribonucleotides (PDRNs) are biomaterials with tissue repair and anti-inflammatory activities, exhibiting multiple activities such as tissue repair, anti-ischemia, and anti-inflammation. Their antioxidant potential has also been gradually revealed in recent years. Certain metal ions (such as zinc and copper) not only act as cofactors for antioxidant enzymes but also exhibit broad-spectrum and mild antibacterial properties. Theoretically, combining PDRNs with specific metal ions could synergistically exert both antioxidant and microecological regulation effects, providing a comprehensive anti-aging solution for the skin, from defense to balance. However, on the one hand, PDRNs, as large molecules, are difficult to transdermally on their own; on the other hand, how to stably and efficiently co-deliver these two highly dissimilar active substances to the target skin layer presents a significant challenge. Simple physical mixing may lead to poor compatibility, instability, loss of activity, or increased skin irritation, failing to achieve the effects of antioxidant activity and skin microecological balance.

[0005] Based on this, the development of microsphere emulsions that can simultaneously carry PDRN and metal ions, stably exert synergistic antioxidant and microecological balance effects, and are suitable for cosmetic systems has significant industrial application value. Summary of the Invention

[0006] This application provides a PDRN-metal ion microsphere emulsion with antioxidant and microecological balancing effects, its preparation method and application, which can achieve highly efficient antioxidant effects and regulate the skin microecology. It aims to solve the technical problems of low antioxidant performance and single function caused by low transdermal permeability of existing PDRN series cosmetics.

[0007] To achieve the above objectives, the present application adopts the following technical solution.

[0008] The first aspect of this application provides a method for preparing a PDRN-metal ion microsphere emulsion with antioxidant and microecological balancing effects, comprising:

[0009] S1, prepare PDRN aqueous solution and metal salt aqueous solution;

[0010] S2, add the metal salt aqueous solution dropwise to the PDRN aqueous solution and stir to obtain a PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects.

[0011] Preferably, the metal salt includes any one of zinc salt, magnesium salt, copper salt, or iron salt.

[0012] More preferably, the metal salt is any one of zinc acetate, zinc gluconate, magnesium acetate, copper nitrate, or ferric chloride.

[0013] Preferably, the concentration of the PDRN aqueous solution is 1 mg / mL;

[0014] The concentration of the aqueous solution of the metal salt is 30 mg / mL.

[0015] Preferably, the mass ratio of PDRN to metal salt is 1:(1~6).

[0016] Preferably, the temperature in S2 is room temperature, and the stirring time is 0.5~3h.

[0017] Preferably, the pH of the PDRN-metal ion microsphere emulsion is 5-8.

[0018] A second aspect of this application provides a PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects prepared by the above preparation method.

[0019] A third aspect of this application provides the application of the above-mentioned PDRN-metal ion microsphere emulsion in cosmetics.

[0020] Compared with the prior art, the beneficial effects of this application are as follows:

[0021] This application achieves a dual technological breakthrough by stably combining PDRN with specific metal ions and integrating them into an emulsion system using microsphere technology. In terms of efficacy, it not only significantly enhances antioxidant performance by activating endogenous antioxidant pathways through PDRN and directly scavenging free radicals with metal ions, but also innovatively utilizes the mild antibacterial properties of metal ions to synergistically regulate the skin microecology, reducing oxidative damage exacerbated by microbial imbalance from the source, thus constructing a dual-dimensional anti-aging mechanism of "scavenging free radicals and regulating the microecology." Moreover, metal ions can synergistically promote PDRN-induced fibroblast migration and angiogenesis through the integrin signaling pathway, forming a positive cycle of "anti-inflammatory and repair." This synergistic effect of the dual mechanisms allows the complex to accelerate tissue remodeling while inhibiting inflammatory outbreaks, providing a more promising intervention strategy for sensitive skin repair or postoperative wound healing. In addition, the microsphere structure not only solves the problems of compatibility stability and transdermal absorption of active ingredients, but also ensures its long-lasting activity in complex emulsion matrices. Ultimately, it yields an innovative cosmetic raw material with excellent antioxidant, anti-inflammatory, repairing, and microecological balancing functions, as well as high stability and superior skin feel, meeting the market's urgent demand for multifunctional integrated skincare formulas. Attached Figure Description

[0022] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments recorded in this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 This is a TEM image of the PDRN-metal ion microsphere emulsion of Example 1 of this application;

[0024] Figure 2 The graph shows the test results of DPPH free radical scavenging ability of the PDRN-metal ion emulsion of Example 1 and the PDRN solution of the comparative example in this application.

[0025] Figure 3 The graph shows the OD value test results of the PDRN-metal ion microsphere emulsion of Example 1 of this application and the PDRN solution of the comparative example against Staphylococcus aureus.

[0026] Figure 4 The graph shows the OD value test results of the PDRN-metal ion microsphere emulsion of Example 1 of this application and the PDRN solution of the comparative example against Propionibacterium acnes. Detailed Implementation

[0027] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this application. All other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are within the scope of protection of this application.

[0028] In the following description of this embodiment, the terms "including", "comprising", "having", and "containing" are all open-ended terms, meaning that they include but are not limited to.

[0029] In the following description of this embodiment, the term "and / or" is used to describe the association relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, B existing alone, and A and B existing simultaneously. A and B can be singular or plural. The character " / " generally indicates that the preceding and following related objects have an "or" relationship.

[0030] In the following description of this embodiment, the term "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions refer to any combination of these items, including any combination of single or multiple items. For example, "at least one of a, b, or c", or "at least one of a, b, and c", can both mean: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, and c can be single or multiple.

[0031] The terminology used in the embodiments of this application is for the purpose of describing particular embodiments only and is not intended to be limiting of this application. The singular forms "a" and "the" as used in the embodiments of this application and the appended claims are also intended to include the plural forms, unless the context clearly indicates otherwise.

[0032] Those skilled in the art should understand that, in the following description of the embodiments of this application, the sequence of numbers does not imply the order of execution. Some or all steps may be executed in parallel or sequentially. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of this application.

[0033] Those skilled in the art will understand that the numerical ranges in the embodiments of this application should be understood as each intermediate value between the upper and lower limits of the specifically disclosed range. Each smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this application. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise stated, the technical / scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. While this application describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this application. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0035] In cosmetic research and development, nanotechnology has great potential for application and product development; metal nano-ions have antioxidant properties, effectively and gradually eliminate free radical cations, have moisturizing properties, and also exert a whitening effect by inhibiting the mRNA expression level of tyrosinase and melanin-related transcription factors.

[0036] This application describes the preparation of PDRN-metal ion microsphere emulsions using supramolecular assembly technology. The preparation method includes:

[0037] S1, prepare PDRN aqueous solution and metal salt aqueous solution;

[0038] In this application, the metal salt is selected from any one of zinc salt, magnesium salt, copper salt or iron salt, and more preferably from any one of zinc acetate, zinc gluconate, magnesium acetate, copper nitrate or ferric chloride.

[0039] The concentration of the PDRN aqueous solution is 1 mg / mL; the concentration of the metal salt aqueous solution is 30 mg / mL.

[0040] S2, add the metal salt aqueous solution dropwise to the PDRN aqueous solution at room temperature and stir for 0.5~3h to obtain a PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects, with a pH of 5~8.

[0041] In this application, the mass ratio of PDRN to metal salt is preferably 1:(1~6).

[0042] This application achieves a dual technological breakthrough by stably combining PDRN with specific metal ions and integrating them into an emulsion system using microsphere technology. In terms of efficacy, it not only synergistically enhances antioxidant performance by activating endogenous antioxidant pathways through PDRN and directly scavenging free radicals with metal ions, but also innovatively utilizes the mild antibacterial properties of metal ions to synergistically regulate the skin's microecology, reducing oxidative damage exacerbated by microbial imbalance at its source, thus constructing a dual-dimensional anti-aging mechanism of "free radical scavenging - microecological regulation." Furthermore, metal ions can synergistically promote PDRN-induced fibroblast migration and angiogenesis through the integrin signaling pathway, forming a positive cycle of "anti-inflammatory - repair." This synergistic effect of the dual mechanisms allows the complex to accelerate tissue remodeling while inhibiting inflammatory outbreaks, providing a more promising intervention strategy for sensitive skin repair or postoperative wound healing.

[0043] In formulation applications, the microsphere structure not only solves the problems of compatibility stability and transdermal absorption of active ingredients, but also ensures its long-lasting activity in complex emulsion matrices.

[0044] The PDRN-metal ion microsphere emulsion of this application has excellent antioxidant, repair and microecological balance functions, as well as high stability and excellent skin feel. It is an innovative cosmetic raw material that meets the market's urgent demand for multifunctional integrated skin care formulas and can be used to prepare cosmetics.

[0045] The PDRN-metal ion microsphere emulsion of this application has an extremely small particle size and is transparent to semi-transparent. Due to its small particle size, it exhibits excellent absorption and moisturizing effects in cosmetics, making cosmetics easier to spread and absorb on the skin. Furthermore, metal ions can inhibit bacteria, reduce UV-induced cell and gene damage, and enhance the skin's antioxidant capacity. Based on these synergistic effects, it intervenes at the root of cellular aging, comprehensively activating the cells' self-repair and regeneration capabilities, and possesses bioactivities such as antioxidant properties, promotion of tissue repair, and delaying aging.

[0046] The PDRN-metal ion microsphere emulsion prepared in this application can improve the stability of PDRN. By co-encapsulating PDRN and metal ions in microspheres, it can protect PDRN from premature degradation by moisture or enzymes in the formulation, while also controlling the release rate of metal ions to avoid potential irritation from the instantaneous release of high-concentration ions. As a large molecule, PDRN is difficult to deliver transdermally on its own; this system enables co-delivery, ensuring that both reach the target action layer simultaneously. Through synergistic effects, the antioxidant and anti-inflammatory effects of both PDRN and metal ions are further amplified.

[0047] The present application will be further illustrated by the following examples.

[0048] Example 1

[0049] S1 Dissolves PDRN in deionized water to prepare 10 mL of PDRN solution with a concentration of 5 mg / mL;

[0050] Zinc gluconate was dissolved in deionized water to prepare a zinc gluconate solution with a concentration of 30 mg / ml.

[0051] S2, take 1.5 mL of zinc gluconate solution and slowly add it dropwise to 10 mL of PDRN solution. Stir at room temperature for 3 h to obtain a milky white, semi-transparent PDRN-zinc ion microsphere emulsion.

[0052] Example 2

[0053] S1, Dissolve PDRN in deionized water to prepare 10 mL of PDRN solution with a concentration of 5 mg / mL;

[0054] Magnesium acetate was dissolved in deionized water to prepare a magnesium acetate solution with a concentration of 30 mg / ml;

[0055] S2, take 3 mL of magnesium acetate solution and slowly add it dropwise to 10 mL of PDRN solution. Stir at room temperature for 3 h to obtain a milky white, semi-transparent PDRN-magnesium ion microsphere emulsion.

[0056] Example 3

[0057] S1, Dissolve PDRN in deionized water to prepare 10 mL of PDRN solution with a concentration of 1 mg / mL;

[0058] Dissolve copper nitrate in deionized water to prepare a copper nitrate solution with a concentration of 30 mg / ml;

[0059] S2, take 100 μL of copper nitrate solution and slowly add it dropwise to 10 mL of PDRN solution. Stir at room temperature for 3 h to obtain a milky white, semi-transparent PDRN-copper ion microsphere emulsion.

[0060] Example 4

[0061] S1, Dissolve PDRN in deionized water to prepare 10 mL of PDRN solution with a concentration of 1 mg / mL;

[0062] Dissolve ferric chloride in deionized water to prepare a ferric chloride solution with a concentration of 30 mg / ml;

[0063] S2, take 50 μL of ferric chloride solution and slowly add it dropwise to 10 mL of PDRN solution. Stir at room temperature for 3 h to obtain a milky white, semi-transparent PDRN-ferric ion microsphere emulsion.

[0064] Comparative Example

[0065] Dissolve PDRN in deionized water to prepare a PDRN solution with a concentration of 4.3 mg / mL.

[0066] The PDRN-zinc ion microsphere emulsion prepared in Example 1 of this application was subjected to TEM testing, and the results are as follows: Figure 1 As shown. From Figure 1 It can be seen that the PDRN-zinc ion microsphere emulsion consists of spherical particles with a relatively uniform distribution.

[0067] The DPPH free radical scavenging ability of the PDRN-zinc ion microsphere emulsion prepared in Example 1 of this application and the PDRN solution prepared in the comparative example was tested. The specific test method is as follows:

[0068] Preparation of DPPH solution: Weigh 1 mg of DPPH using a precision balance, dissolve it in anhydrous ethanol by sonication, and bring the volume to 20 ml. Protect from light and use immediately.

[0069] Preparation of zinc gluconate solution: Weigh zinc gluconate and dissolve it in deionized water to prepare a zinc gluconate solution with a concentration of 3.9 mg / ml.

[0070] In a 96-well plate, blank tubes, test tubes, control tubes, and blank control tubes were set up respectively. 200 μl of reagent was added to each test tube, as shown in Table 1.

[0071] Table 1. Feeding Table for DPPH Free Radical Scavenging Ability Test

[0072]

[0073] Add the reagents to the 96-well plate according to the operating table, shake to mix, and incubate at room temperature in the dark for 30 minutes. Measure the absorbance at 517 nm using a microplate reader. Each assay tube should have a corresponding control tube. Blank tubes only need to be measured 1-2 times.

[0074] Experimental data processing:

[0075] All data were calculated using the following formulas, and graphs were generated using Excel software. The calculation formulas are as follows:

[0076] DPPH radical scavenging rate P% = (1 - (As - Ac) / (Ab - A empty)) 100%

[0077] P% – Clearance rate;

[0078] As — the absorbance of the mixture of the test solution and DPPH solution;

[0079] Ac—the absorbance of the mixture of the test solution and anhydrous ethanol solution;

[0080] Ab—Absorbance of the mixture of DPPH solution and sample solvent solution;

[0081] A_blank — Absorbance of the blank tube.

[0082] DPPH free radical scavenging rate test results are as follows Figure 2 As shown. From Figure 2 It can be seen that the DPPH free radical scavenging rate of Comparative Example 1 was 51.3%, the DPPH free radical scavenging rate of zinc gluconate was 3.6%, and the DPPH free radical scavenging rate of Example 1 was 80.4%. This shows that the introduction of metal ions significantly enhanced the free radical scavenging ability of PDRN. Moreover, it is not a simple superposition of effects, but rather a demonstration that PDRN and metal ions produced a "1+1>2" synergistic antioxidant effect.

[0083] The antibacterial properties of the PDRN-zinc ion microsphere emulsion prepared in Example 1 of this application and the PDRN solution prepared in the comparative example were tested. The specific methods are as follows:

[0084] Culture medium preparation: Measure 100 mL of distilled water using a graduated cylinder and pour it into a 250 mL reagent bottle. Weigh 2.5 g of LB broth culture medium using an analytical electronic balance, add the broth, mix well, and sterilize in a high-temperature, high-pressure steam sterilizer at 121°C for 15 min. The LB broth culture medium was purchased from Shanghai Bioengineering Co., Ltd., model A507002.

[0085] Preparation of bacterial suspension: Take two 12 mL bacterial culture tubes and add 3 mL of LB liquid medium to each. Pick a single colony from the solid medium of Staphylococcus aureus and Propionibacterium acnes and add it to the liquid medium. Use the other tube as a blank control. Place both bacterial culture tubes in a constant temperature shaker (37℃, 200 rpm) and shake overnight (15 h). Wash off the colonies with LB liquid medium and adjust the bacterial concentration to 10. 6 CFU / mL.

[0086] Sample preparation: Take an appropriate amount of the original test solution sample and place it into a 12 mL bacterial culture tube according to the group. Sterilize under ultraviolet light for 60 min and set aside. The experimental groups are set as control group, comparative example group 1, and example 1 group.

[0087] OD value test: Dilute the bacterial culture to a concentration of 10⁶ CFU / mL using LB broth medium.

[0088] Among them, the sample group: 1 ml of the sample from Comparative Example 1 and Example 1 was added to the corresponding 12 ml bacterial culture tube, and then 1 ml of diluted bacterial solution was added to the tube.

[0089] Control group: 1 ml of sterile PBS solution was added to 1 ml of diluted bacterial solution.

[0090] The Staphylococcus aureus group was incubated statically at 37°C for 18 hours; the Propionibacterium acnes group was incubated anaerobically at 37°C for 48 hours. After incubation, 200 μL of co-culture medium was added to each well of a 96-well plate, and the OD values ​​of each group were measured at 600 nm using a microplate reader. The data were visualized using GraphPad Prism 8.0.

[0091] OD value test results as follows Figure 3 and Figure 4 As shown. Figure 3 The OD value test results for the sample against Staphylococcus aureus are from... Figure 3 It can be seen that the sample in Example 1 has a certain antibacterial effect against Staphylococcus aureus, while Comparative Example 1 has no obvious antibacterial effect against Staphylococcus aureus.

[0092] Figure 4 The OD value test results for the sample against Propionibacterium acnes are from... Figure 4 It can be seen that the sample of Example 1 has a certain antibacterial effect against Propionibacterium acnes, while the sample of Comparative Example 1 has no obvious antibacterial effect against Propionibacterium acnes.

[0093] Although this application has been described in detail in this specification with general descriptions and specific embodiments, some modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, such modifications or improvements made without departing from the spirit of this application are all within the scope of protection claimed in this application.

Claims

1. A method for preparing a PDRN-metal ion microsphere emulsion with antioxidant and microecological balancing effects, characterized in that, include: S1, prepare PDRN aqueous solution and metal salt aqueous solution; S2, add the metal salt aqueous solution dropwise to the PDRN aqueous solution and stir to obtain a PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects.

2. The preparation method according to claim 1, characterized in that, The metal salt includes any one of zinc salt, magnesium salt, copper salt, or iron salt.

3. The preparation method according to claim 2, characterized in that, The metal salt is any one of zinc acetate, zinc gluconate, magnesium acetate, copper nitrate, or ferric chloride.

4. The preparation method according to claim 1, characterized in that, The concentration of the PDRN aqueous solution is 1 mg / mL; The concentration of the aqueous solution of the metal salt is 30 mg / mL.

5. The preparation method according to claim 1, characterized in that, The mass ratio of PDRN to metal salt is 1:(1~6).

6. The preparation method according to claim 1, characterized in that, The temperature in S2 is room temperature, and the stirring time is 0.5~3h.

7. The preparation method according to claim 1, characterized in that, The pH of the PDRN-metal ion microsphere emulsion is 5-8.

8. The PDRN-metal ion microsphere emulsion with antioxidant and microecological balance effects prepared by the preparation method according to any one of claims 1-7.

9. The application of the PDRN-metal ion microsphere emulsion according to claim 8 in cosmetics.

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