Biomimetic anti-aging skincare composition with prebiotic, postbiotic and pro biotic enriched components

Abstract

The invention relates to a biomimetic anti-aging skincare composition comprising: a liposomal blend having a mixture of: (i) one or more prebiotics; (ii) one or more probiotics; and (iii) one or more

Description

FIELD OF THE INVENTION

[0001] The present invention relates to biomimetic anti-aging skincare compositions comprising liposomally encapsulated prebiotics, probiotics, and postbiotics. More particularly, the invention pertains to a method for preparing the composition that mimic the skin's natural structure and microbiome to enhance hydration, elasticity, and barrier function. BACKGROUND OF THE INVENTION

[0002] Human skin is continuously exposed to environmental stressors such as temperature fluctuations, pollution, ultraviolet radiation, and emotional stress. These factors compromise the skin’s natural barrier, leading to dehydration, sensitivity, dullness, and visible signs of aging such as fine lines and loss of elasticity. Biomimetism offers a targeted solution by imitating the skin’s natural composition and processes. Biomimetic skincare leverages high-tolerance, natural-origin ingredients that closely resemble components naturally found in the skin such as water, lipids, proteins, sugars, and mineral salts. As a result, these formulations are better accepted by the skin and support its innate regenerative functions.

[0003] The term “biomimicry” stems from “bio,” meaning life, and “mimetic,” meaning to imitate. It refers to the scientific practice of studying nature’s systems and translating them into functional solutions for human challenges. In skincare, biomimetic formulations mimic the architecture and chemistry of healthy skin, enabling optimal absorption, enhanced efficacy, and improved compatibility. Modem biomimetic skincare represents a convergence of nature and science, using both plant-derived and synthetic analogs that replicate biological structures and pathways. This approach offers not only superior performance but also supports sustainability and ethical product development, eliminating the need for animal testing and favoring cruelty-free, eco-conscious alternatives.

[0004] The global skin care products market size was valued at USD 130.50 billion in 2021 and is expected to expand at a compound annual growth rate (CAGR) of 4.6% from 2022 to 2030. Escalating demand for face creams, sunscreens, and body lotions across the globe is expected to have a positive impact on the market growth over the forecast period. Moreover, the flourishing e-commerce sector is anticipated to boost market growth further. In the UK, revenue in the Skin Care segment amounts to US$4.05bn in 2023. The market is expected to grow annually by 2.38% (CAGR 2023-2027). The organic skincare market is evolving at a rapid pace evident in the growth statistics. The Global Organic Skincare Market size was valued at USD 9.83 billion in 2021 and is poised to grow from USD 10.49 billion in 2022 to USD 21.16 billion by 2030, growing at a CAGR of 8.9% in the forecast period (2023-2030). The revenue of the natural and organic cosmetics market in the United Kingdom amounted to around 221 million British pounds in 2020. That same year, the average revenue per capita stood at 3.25 British pounds. Analysts note that millennials are a key target demographic for organic skincare brands. This generation is more likely to be concerned about environmental issues and their own personal health, making them more likely to purchase organic skincare products.

[0005] Biomimetic skincare is an emerging approach to skincare formulation that sits at the intersection of cutting-edge scientific technology and nature. “These are synthetic or natural materials that can mimic our own skin’s natural function,” explains Ginger King, cosmetic chemist. “Biomimetic materials [are] triggers for our own natural production of collagen, elastin, sebum, etc., instead of applying materials like collagen on skin, which is useless.” Biomimicry has been the catalyst for the discovery of powerful anti-aging peptides, howto reignite slowed collagen production, and superior moisturizing ingredients. Biomimetic skincare doesn’t refer to specific ingredients so much as it refers to a framework from which ingredients and formulas are derived.

[0006] It’s also more sustainable. Natural ingredients aren’t always the most sustainable, and scientists can use biomimetics to formulate new alternatives that are more sustainable with less environmental impact. Thus, the future of beauty lies in the past - millions of years of evolution across all species. Time has proven that nature has always been the cure - biomimetics just harnesses that cure and distils that knowledge into practical applications that solve real, human problems. Also, because biomimetic products use naturally grown ingredients that are free from toxic pesticides and fertilizers, they don't leave a harmful footprint on the planet, particularly the soil, water, and air. Given the growing market demand for organic products and the emerging trend of biomimetics particularly in the skin care industry offers a progressive scope of skin products that improves the health of skin giving it a long-lasting natural impact.

[0007] Despite the proliferation of anti-aging skincare products in the market, many fail to effectively mimic the biological and structural complexity of human skin. Current formulations often lack targeted delivery systems, contain harsh or incompatible ingredients, or provide only superficial benefits. There exists a clear need for an advanced, biocompatible skincare system that can work in synergy with the skin’s microbiome, structure, and function to provide long-lasting, meaningful results in aging-related skin concerns. SUMMARY OF THE INVENTION

[0008] In view of the limitations associated with conventional anti-aging skincare products, the general purpose of the present invention is to provide a biomimetic skincare composition and method of preparation that overcomes the above mentioned shortcomings. The present invention aims to offer a composition that mimick the skin’s natural structure, delivering biologically active components in a stable liposomal system, and promoting hydration, barrier repair, and microbiome balance, all while maintaining high biocompatibility and skin tolerance.

[0009] In light of above limitations, an aspect of the present invention relates to a biomimetic anti-aging skincare composition. The composition includes a liposomal blend encapsulating a mixture, a gel network, and one or more cosmetically acceptable additives. The mixture includes one or more prebiotics, one or more probiotics, and one or more postbiotics. The gel network includes water and a gallant. The one or more cosmetically acceptable additives are selected from the group including emollients, humectants, peptides, ceramides, vitamins, antioxidants, emulsifiers, preservatives, and plant extracts. Herein, the composition is adjusted at a pH in the range of 4.5 to 5.5 to mimic the natural acid mantle of human skin.

[00010] In one embodiment, the one or more prebiotics are selected from oat meal extract, banana extract, asparagus extract and a combination thereof in a range of 0.6 % to 1 % by weight of the total weight of the liposomal blend.

[00011] In one embodiment, the one or more probiotics are selected from Lactobacillus plantarum (KPBBC7), Lactobacillus Arizonensis Ferment Filtrate, Bacillus Ferment, Saccharomyces Ferment Filtrate, Kombucha Tea, Lactobacillus Ferment Lysate, Bacillus Ferment &Saccharomyces Ferment Filtrate or a combination thereof in a range of 0.5 % to 5 % by weight of the total weight of the liposomal blend.

[00012] In one embodiment, the one or more postbiotics are selected from alpha-glucan oligosaccharide, metabolites of Lactobacillus delbrueckii or L. fermentum, or bacterial fermentation supernatant or a combination thereof in a range of 1 % to 2.5 % by weight of the total weight of the liposomal blend.

[00013] In one embodiment, the liposome blend are phospholipid-based vesicles formed using thin-fdm hydration followed by sonication or extrusion to achieve a particle size ranging from 50 to 200 nm.

[00014] In one embodiment, the antioxidant comprises Undaria pinnatifida cell culture extract in an amount ranging from 0.5% to 1.5% by weight.

[00015] In light of above limitations, another aspect of the present invention relates to a method of preparing a biomimetic anti-aging skincare composition. The method includes preparing and heating a water phase and an oil phase at a temperature in a range of 70°C to 80°C. The water phase includes humectants and water-soluble ingredients and the oil phase includes carrier oils, emulsifiers, and oil-soluble ingredients. Combining the oil phase with the water phase under high-shear stirring to form an emulsion. Followed by cooling the emulsion at a temperature in a range of 30°C to 40°C. Further, adding a liposomal blend encapsulating one or more prebiotics, one or more probiotics, and one or more postbiotics to the emulsion under gentle mixing. Adjusting the pH of the final composition to between 4.5 and 5.5 and optionally adding fragrances, preservatives, antioxidants, and additional actives and homogenizing the composition to obtain a stable anti-aging skincare composition.

[00016] In one embodiment, the liposomal blend is prepared by a thin-film hydration process. The process includes dissolving phospholipids and cholesterol in an organic solvent. Followed by evaporating the organic solvent to form a lipid film and hydrating the obtained lipid film with an aqueous solution containing the one or more prebiotics, the one or more probiotics, and the one or more postbiotics. Thereafter, sonicating the lipid film to a desired nanosized liposomal blend.

[00017] In one embodiment, the desired nanosized liposomal blend has a particle size distribution in the range of 50 to 200 nm.

[00018] In one embodiment, the composition is homogenized post-addition of the liposomal blend using a high-shear mixer at a speed of 1,000-3,000 rpm for 5-10 minutes to ensure uniform distribution.

[00019] These elements, together with the other aspects of the present disclosure and various features are pointed out with particularity in the claims annexed hereto and form a part of the present disclosure. For a better understanding of the present disclosure, its operating advantages, and the specified object attained by its uses, reference should be made to the accompanying drawings and descriptive matter in which there are illustrated exemplary embodiments of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[00020] The advantages and features of the present disclosure will become better understood with reference to the following detailed description taken in conjunction with the accompanying drawing, in which:

[00021] Fig. 1 illustrates the Skin elasticity (R2) increase percentage;

[00022] Fig 2 illustrates the Fatigue Resistance (F4) decrement percentage;

[00023] Fig. 3 illustrates the Cell growth increase percentage;

[00024] Fig 4 illustrates the Cell Proliferation rate percentage.

[00025] Fig. 5 illustrates the HABP straining of the vehicle-treated (VEH) human skin explant tissues and the 0.04% stabilized-biomimetic complex (BMC) -treated skin explant tissues;

[00026] Fig 6 illustrates the comparison between the epidermis thickness of the vehicle-treated tissue and the epidermis thickness of the stabilized-biomimetic complex (BMC); and

[00027] Fig 7 illustrates the present Biomimetic anti-aging cream that employs a barrier lesion.

[00028] Like reference numerals refer to like parts throughout the description of several views of the drawing. DETAILED DESCRIPTION OF THE DISCLOSURE

[00029] Embodiments are provided so as to thoroughly and fully convey the scope of the present disclosure to the person skilled in the art. Numerous details are set forth, relating to specific components, and methods, to provide a complete understanding of embodiments of the present disclosure. It will be apparent to the person skilled in the art that the details provided in the embodiments should not be construed to limit the scope of the present disclosure. In some embodiments, well-known processes, well-known apparatus structures, and well-known techniques are not described in detail.

[00030] It will be appreciated that for simplicity and clarity of illustration, where appropriate, reference numerals have been repeated among the different figures to indicate corresponding or analogous elements. In addition, numerous specific details are set forth in order to provide a thorough understanding of the exemplary embodiments described herein. However, it will be understood by those of ordinary skill in the art that the exemplary embodiments described herein can be practiced without these specific details. In other instances, methods, procedures, and components have not been described in detail so as not to obscure the related relevant feature being described. The drawings are not necessarily to scale and the proportions of certain parts may be exaggerated to better illustrate details and features. The description is not to be considered as limiting the scope of the exemplary embodiments described herein. The term “comprising” means “including, but not necessarily limited to”; it specifically indicates open-ended inclusion or membership in a so-described combination, group, series, and the like.

[00031] The terminology used, in the present disclosure, is only for the purpose of explaining a particular embodiment and such terminology shall not be considered to limit the scope of the present disclosure. As used in the present disclosure, the forms "a," "an," and "the" may be intended to include the plural forms as well, unless the context clearly suggests otherwise. The terms "comprises," "comprising," "including," and "having," are open-ended transitional phrases and therefore specify the presence of stated features, integers, steps, operations, elements, modules, units, and / or components, but do not forbid the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. The particular order of steps disclosed in the method and process of the present disclosure is not to be construed as necessarily requiring their performance as described or illustrated. It is also to be understood that additional or alternative steps may be employed.

[00032] The present invention relates to a biomimetic anti-aging skincare composition and a method for preparing. The biomimicry is most impactful and visible in skincare. A truly biomimetic formulation will include the three key lipids including ceramides, cholesterol, and free fatty acids in a ratio similar to that found naturally in the skin (e.g., 3:1:1 or 1:1:1 for Ceramide:Cholesterol:Free Fatty Acid). These are crucial for rebuilding and reinforcing the skin's natural barrier. The advanced formulations use specific emulsifiers (e.g., lecithin, hydrogenated phospholipids) and lipid blends that can self-assemble into liquid crystal or lamellar structures when mixed with water. This mimics the natural arrangement of lipids in the stratum comeum, creating a "second skin" that seamlessly integrates with the existing skin barrier. This lamellar structure allows for enhanced barrier function that fills in gaps in a compromised barrier, reducing trans-epidermal water loss (TEWL), optimal delivery i.e. the lamellar layers can encapsulate and gradually release active ingredients, facilitating their penetration into the epidermis, and increased compatibility as the structure is akin to the skin's own, it's highly compatible and less likely to cause irritation.

[00033] The natural moisturizing factors (NMFs) include components such as: amino acids that often found in protein hydrolysates or as individual amino acids, urea which is considered as a powerful humectant and keratolytic agent, PCA (Pyrrolidone Carboxylic Acid) which is considered as a naturally occurring humectant, Hyaluronic Acid (various molecular weights): Low molecular weight HA can penetrate deeper for hydration, while high molecular weight HA forms a film on the surface, mimicking the skin's natural ability to attract and hold water, and Keratinocyte Mimicry (Indirect): While the formulation doesn't contain live keratinocytes, ingredients like peptides can act as signaling molecules (biomimetic peptides) to stimulate the skin's own keratinocytes to produce more collagen, elastin, or other beneficial components, or to enhance their differentiation and barrier formation.

[00034] Mimicking the Dermis and Extracellular Matrix (ECM): While topical products have limited penetration into the dermis, biomimetic formulations can still influence its health. Specific biomimetic peptides are designed to mimic fragments of natural collagen or elastin, or to stimulate fibroblasts to produce more collagen and elastin, thereby improving skin firmness and elasticity. Some formulations incorporate biomimetic growth factors that can signal to dermal cells to initiate repair and regeneration processes. While HA is also an NMF in the epidermis, its primary role is in the dermal ECM. Formulations use various molecular weights of HA to provide hydration and plumping effects, mimicking the skin's natural waterholding capacity in the dermis. Ingredients like Vitamin C (L-Ascorbic Acid) and Vitamin E, or botanical extracts like Undaria pinnatifida, help protect dermal components (collagen, elastin) from oxidative damage, mimicking the skin's natural defense mechanisms.

[00035] Mimicking Skin's Microbiome (via Pre-, Pro-, Post-biotics): This is a newer and highly sophisticated aspect of biomimetic skincare. Prebiotics are "food" for beneficial skin bacteria (e.g., oligosaccharides, inulin). By including prebiotics, the formulation supports the growth of a healthy microbiome on the skin's surface, mimicking the natural symbiotic relationship between the skin and its resident microorganisms. Probiotics: While often difficult to formulate as live organisms, some advanced formulations (often encapsulated in liposomes, as detailed earlier) deliver beneficial bacterial strains to the skin. This directly introduces "good" bacteria to help balance the skin's natural flora, which is crucial for barrier function, immune response, and overall skin health. Postbiotics: These are the beneficial metabolic byproducts of probiotics (e.g., lactic acid, peptides, enzymes, hyaluronic acid fragments). By directly applying postbiotics, the formulation delivers the functional benefits of a healthy microbiome, such as anti-inflammatory, antimicrobial, and barrier-strengthening effects, without needing live bacteria. This directly mimics the positive impact of a balanced skin microbiota.

[00036] Healthy skin has a slightly acidic pH (around 4.5-5.5), known as the "acid mantle." Biomimetic formulations are carefully pH-adjusted to this range, as it is crucial for the following like enzyme activity, microbiome balance, and barrier integrity. A truly biomimetic skincare composition doesn't just "add" ingredients; it designs the entire product, including its structure, ingredient ratios, and delivery systems, to work in harmony with the skin's inherent biology. By mimicking the skin's lipid matrix, NMFs, ECM components, and even its microbiome and pH, these formulations aim to enhance the skin's natural functions of protection, hydration, regeneration, and repair, leading to a healthier, more resilient, and more youthful complexion.

[00037] The composition includes a liposomal blend encapsulating a mixture having one or more prebiotics, one or more probiotics, and one or more postbiotics, a gel network, and one or more cosmetically acceptable additives. The gel network includes water and a gallant. The one or more cosmetically acceptable additives are selected from the group including emollients, humectants, peptides, ceramides, vitamins, antioxidants, emulsifiers, preservatives, and plant extracts. The proposed mixture contains prebiotic, postbiotic, and probiotic ingredients in such a unique and optimized ratio that it balances and nourish the skin microbiome (trillions of microorganisms, like bacteria, fungi, and viruses found on the skin as invisible lifeforms are known as the skin microbiome).

[00038] The one or more prebiotics, one or more probiotics, and one or more postbiotics are encapsulated into liposomes for use in skincare formulations. This encapsulation is a highly advanced delivery system that offers significant advantages in terms of stability, targeted delivery, and efficacy, especially for sensitive or live ingredients like probiotics.

[00039] Liposomes shield these sensitive components from degradation by environmental factors (oxygen, light, temperature fluctuations), harsh formulation ingredients (preservatives, high pH), and even the skin's own enzymes. This significantly improves the shelf-life and viability of probiotics and the integrity of pre- and postbiotics. Liposomes, with their lipid bilayer structure mimicking cell membranes, and may readily fuse with or be absorbed by skin cells, facilitating deeper penetration and more efficient delivery of the encapsulated ingredients to their target sites within the skin. Liposomal encapsulation enables a sustained and controlled release of the encapsulated substances, prolonging their activity. By encapsulating potentially irritating ingredients, liposomes can help minimize adverse reactions on sensitive skin.

[00040] In one embodiment, the liposomal blend is prepared by a thin-film hydration process. The process includes dissolving phospholipids and cholesterol in an organic solvent to obtain a lipid solution. The primary components of liposomes are phospholipids (e.g., phosphatidylcholine from soy or egg lecithin). The choice of lipids (type, chain length, saturation, charge) influences the liposome's fluidity, stability, and encapsulation efficiency. Cholesterol is often included to provide rigidity and reduce permeability. The organic solvent is selected from chloroform, ethanol, methanol, or mixtures thereof. The obtained lipid solution is placed in a round-bottom flask. The organic solvent is then removed under vacuum (e.g., using a rotary evaporator) to form a thin, dry lipid film on the inner surface of the flask. This film consists of a disordered array of lipids. A biomimetic blend containing one or more prebiotic, one or more probiotic and one or more postbiotic ingredients are prepared in an aqueous solution. The water-soluble prebiotics (e.g., inulin, FOS, GOS) are dissolved in water. The live probiotic strains are prepared as a concentrated suspension in a compatible buffer, often with cryoprotectants if they were freeze-dried. Maintaining their viability is paramount. The bioactive compounds (peptides, organic acids, enzymes) derived from probiotic fermentation are dissolved in water. The aqueous solution of the biomimetic blend containing the prebiotic, probiotic, or postbiotic is added to the dried lipid film. The flask is gently rotated or agitated above the lipid phase transition temperature (to ensure fluidity of the lipids) to allow the lipid film to hydrate and spontaneously form multi-lamellar vesicles (MLVs). During hydration, the aqueous solution becomes trapped within the forming lipid bilayers. The MLVs are often heterogeneous in size and can be quite large. To produce smaller, more uniform, and often unilamellar vesicles (SUVs or LUVs) for better skin penetration and stability, further processing is needed. High-frequency sound waves are applied to the liposome dispersion, breaking down large vesicles into smaller ones. The liposome dispersion is passed multiple times through polycarbonate membranes with defined pore sizes (e.g., 50-200 nm). This forces the vesicles to reform at the desired size. The dispersion is subjected to high pressure through a narrow orifice, leading to size reduction.

[00041] The present invention relates to the method for preparing a biomimetic antiaging skincare composition. The method is performed in four stages comprising a preformulation and preparation stage, heating and emulsification stage, cooling stage, and postformulation stage.

[00042] In the Pre-Formulation and Preparation stage, all raw materials (oils, waters, emulsifiers, thickeners, active ingredients, preservatives, fragrances, and the biomimetic blend) are sourced from reputable suppliers and undergo rigorous Quality Control (QC) checks to ensure purity, potency, and absence of contaminants. All glassware, mixing vessels, stirrers, and other equipment are thoroughly cleaned, sanitized, and sterilized to prevent contamination. This if followed by a weighing and pre-mixing stage where water phase ingredients (Phase A), oil phase ingredients (Phase B) and a Biomimetic Blend (Phase C) are weighted and prepared. The water phase ingredients include water, humectants (e.g., glycerin, hyaluronic acid), watersoluble extracts, and water-soluble preservatives are accurately weighed into a clean vessel. The oil Phase Ingredients (Phase B) includes carrier oils, butters, fatty alcohols, waxes, oilsoluble emulsifiers, and oil-soluble active ingredients (if any) are weighed into a separate vessel. The heat-sensitive actives &biomimetic blend (Phase C) ingredients (e.g., certain vitamins, peptides, probiotics), are kept separate and at room temperature or chilled, as per supplier recommendations. These typically come pre-solubilized or as a suspension in a compatible solvent.

[00043] In the Heating and Emulsification stage, the vessel containing the water phase is heated to a specified temperature (e.g., 70-80°C) with continuous stirring, ensuring all ingredients are fully dissolved. Simultaneously, the vessel containing the oil phase is heated to the same temperature (e.g., 70-80°C). This ensures both phases are at a similar temperature, facilitating proper emulsification. Stirring is maintained to ensure uniform melting and mixing of oil-soluble components. Once both phases reach the target temperature and are homogenous, the oil phase (B) is slowly added to the water phase (A) with continuous, high-shear stirring. This is a critical step where the emulsion is formed. The high shear helps to create small, uniform oil droplets dispersed within the water phase (or vice versa, depending on the emulsion type, O / W or W / O). Stirring continues until a stable emulsion forms. After emulsification, the mixture is allowed to cool slowly with continued, gentler stirring. This helps the emulsion to stabilize and thicken.

[00044] In the Cooling Phase, the addition of biomimetic blend and other sensitive actives are added. As the emulsion cools to a temperature below 40°C (ideally around 30-35°C, or as specified by the biomimetic blend supplier), the heat-sensitive active ingredients and the biomimetic blend (Phase C) are slowly and carefully added to the main emulsion. This is followed by gentle, continuous mixing to ensure even dispersion of the biomimetic blend without disrupting the emulsion or damaging the delicate components. Homogenization might be suggested for some biomimetic blends, but this would typically occur after their addition and still at a lower temperature to ensure even distribution. The pH of the final product is checked and adjusted to the desired range (often skin-compatible, e.g., 4.5-6.5) using appropriate acids (e.g., lactic acid, citric acid) or bases (e.g., sodium hydroxide solution). This is done after the addition of the biomimetic blend to ensure its stability within the final pH range. Remaining preservatives (if not already added in earlier phases) and fragrance components are added at this stage, again to minimize exposure to high heat.

[00045] In the Post-Formulation stage, the product may undergo a final homogenization step (e.g., using a homogenizer or high-shear mixer) at a lower speed to refine the particle size and ensure consistency. The finished product undergoes a comprehensive QC process, including: Appearance: Visual inspection for color, texture, and uniformity, pH: Confirmation of the final pH within specifications, Viscosity: Measurement of consistency, Stability Testing: Accelerated aging tests (e.g., heat / cold cycles, centrifuge stability) and long-term shelf-life studies, Microbiological Testing: To ensure the product is free from harmful microorganisms, Active Ingredient Assay: To confirm the concentration of key active ingredients. Once all QC parameters are met, the product is filled into appropriate packaging (bottles, jars, tubes) under hygienic conditions. By adding the biomimetic blend in the cooling phase, the formulator ensures that its delicate structure and biological activity are preserved, allowing it to function optimally in mimicking the skin's natural processes.

[00046] Anti- Aging Biomimetic Complex Efficacy evaluation: The test method was carried out by connecting an elastic probe PVM600 manufactured by Courage+ Khazaka, Germany (CK, Germany) to an MPA master according to a standard method and using the probe in combination with Cutomer Q analysis software.

[00047] Working principle of skin elasticity test probe: The test principle is based on the principle of suction and stretching, where a negative pressure is generated on the skin surface to be tested to suck the skin into a specific test probe, and the depth of the skin sucked into the test probe is measured by a non-contact optical test system. The test probe includes a light emitter and a light receiver, the ratio of light (the ratio of emitted light to received light) is proportional to the depth of skin being absorbed, thus obtaining a curve of the length of skin stretched versus time, which is then analyzed by the MPA software to determine the elastic properties of the skin.

[00048] Technical parameter index of skin elasticity test probe

[00049] Host physical size: 27 X 25.5 X7 cm, probe: phi 2X10cm Probe test hole diameter: phi 2mm

[00050] Pressure: (20-500) mbar Test time: (0.1-60) seconds

[00051] Precision: penetration depth of about 3% above lOOum, and wireless property below lOOum Power supply: 100 to 240V / 50-60Hz / 0.3A

[00052] Interface: USB (universal serial bus)

[00053] Test negative pressure mode: continuous negative pressure mode Curve display mode: stretch length-time table

[00054] Skin elasticity test mode-maintaining constant negative pressure

[00055] Uf=Ue+Uv

[00056] Wherein: uf-maximum skin stretches with negative pressure.

[00057] Ue — the amount of skin stretches at 0.1 second after a constant negative pressure is applied to the skin, the amount of stretch in the elastic portion is located.

[00058] Uv-Uf-Ue is the amount of stretching of the viscoelastic part, or plastic part, of the skin.

[00059] The value of Ue is higher for skin that is more elastic than for younger skin, whereas the value of Ue is lower for skin that is less elastic than for older skin, and the value of Uv is higher for viscoelastic.

[00060] Ur=Uf-Ul.lUa=Uf-U2.0

[00061] Wherein: Ur-the value of elastic part Ur, i.e. the value of elastic part and the value of viscoelastic part, also called plastic part, after negative pressure is removed for 0.1 second.

[00062] Ua-the recovery value of the skin from the removal of negative pressure to the next successive test of the skin surface plus negative pressure.

[00063] The younger the skin, the more elastic the skin, the higher the elastic fraction value Ur; the older the skin, the less elastic the skin, the lower the Ur value.

[00064] Skin elasticity test method

[00065] Opening the Curometer Q software, clicking the tool bar Measurement or directly clicking the upper right Measurement button, taking down the probe cover after the pump is started, clicking OK, vertically and lightly contacting the probe with the skin area to be tested within 30 seconds (the skin surface should be cleaned, if a product is smeared, the test is carried out after the product is absorbed, so as not to pollute the probe hole), starting the test until the test cycle time is over, moving the probe away, and covering the probe cover. The 3-point test was performed in the area 2.5 X 2.5cm from the arm's upper arm wrist, and the average value of R2 was calculated, R2 = Ua / Uf. Meanwhile, the anti-fatigue degree value F4 of the skin can be calculated according to the measured data. Then, after the area was covered with a mask cloth, 0.45ml of the sample to be tested was injected into the film cloth, the film cloth was removed after applying for 20 minutes, the mask liquid was washed off after waiting for 10 minutes, and the skin elasticity value R2 and the fatigue resistance degree value F4 of the skin were measured after the skin was dried.

[00066] Fibroblast assay

[00067] Cell counting method: After cell suspension preparation, cells are stained with the vital dye trypan blue for cell counting. Trypan blue is unable to penetrate the normal intact cell membrane of living cells, so living cells are not stained. The cell membrane permeability of the dead cells is increased, and the dye enters the cells to stain the cells (blue).

[00068] MTT staining: MTT is named as 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2-H-tetrazolium bromide, and the Chinese chemical name is 3- (4, 5-dimethylthiazole-2) -2,5-diphenyl tetrazolium bromide, the trade name is: thiazole blue, a yellow dye. The MTT method is also called MTT colorimetric method, and is a method for detecting cell survival and growth. The detection principle is that succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT into water-insoluble blue- purple crystalline Formazan (Formazan) and deposit the blue-purple crystalline Formazan in the cells, and dead cells do not have the function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and its light absorption value is measured at 540 or 720nm wavelength by ELISA detector, which can indirectly reflect living cell number. Within a certain range of cell number, MTT crystals are formed in an amount proportional to the cell number.

[00069] Test materials and methods: Experimental materials DMEM, DPBS, Typsin-EDTA, Petri dish, 96well dish, 0.4% trypan blue solution, absolute ethyl alcohol or 95% ethyl alcohol solution, common microscope, cell counting plate, pipette, MTT, DMSO

[00070] The experimental method comprises the following steps:

[00071] Fibroblast cell culture: Treating cultured fibroblast with Typsin-EDTA, collecting, suspending with DMEM, counting with a hemocytometer, and diluting to 5 x 10 cell concentration4cells / ml for use. The prepared cell suspension was aliquoted into 96well dish and 6well dish, respectively, and cultured, wherein the inoculation amount of 96well dish was lOOul, and the inoculation amount of 6well dish was 2 ml. The cells were incubated at 37 °C in an incubator containing 5% CO2 for 24 hours.

[00072] Sample addition

[00073] (1) Diluting a sample to be detected by using a DMEM culture medium, wherein the diluted concentrations are as follows: 1% (this concentration passes the previous MTT test and is non-toxic)

[00074] (2) After 24 hours of cell culture, the previous DMEM was removed, and then carefully washed with DPBS (3) the DMEM medium to which the sample to be tested was added prepared in the first step is added in order. The cells were incubated at 37 °C in an incubator containing 5% CO2 for 48 hours.

[00075] Cell counting

[00076] Counting plate treatment: Wiping the counting plate with absolute ethyl alcohol or 95% ethyl alcohol solution, wiping the counting plate with silk cloth, wiping another cover glass sheet, and covering the cover glass on the counting plate.

[00077] Dyeing process: The cells cultured in 6well dish were removed from the medium, digested with typsin-EDTA, collected and suspended in DMEM medium. lOul of 0.4% trypan blue stain and lOul of cell suspension were pipetted with a pipette and mixed well. Slowly injecting from the edge of the counting plate to fill the gap between the counting plate and the cover plate. The counting plate was placed under a low power mirror (10X 10 times) to observe the counting.

[00078] Counting method: The number of cells in the four large squares (16 small squares per large square) of the counting plate was counted as shown. When counting, only intact cells are counted, and if cells are aggregated, the cells are counted as one cell. In a large square, if any cells are on-line, the off-line cells are generally counted without the on-line cells, and the left-line cells are counted without the right-line cells. Under the observation of the microscope, the cells with strong refractivity and no coloration are live cells, and the cells with blue coloration are dead cells.

[00079] Conversion of the count: After counting, the number of cells per ml of suspension was converted. Since the area of each square in the counting plate is 0.01cm2 Height of 0.01cm, so that its volume is 0.0001cm3I.e. 0.1mm3. Since 1ml is 1000mm3Therefore, the number of cells in each large square x 10000 = cell number / ml, and thus can be calculated as follows: cell suspension cell number / ml-4 big lattice cell total number / 4 x 10000. If the sample is diluted before counting, the dilution factor can be multiplied. After counting the cells, the concentration of the cells in the cell suspension is calculated. The cell proliferation rate of each sample was calculated based on the blank.

[00080] MTT staining: The MTT powder stored in the frozen state was dissolved in DPBS to prepare a solution having a concentration of 2 mg / ml. After 48 hours of culture in 96well dish, 50ul / well of MTT solution was added after removing the medium. Culturing at 37 deg.C in 5% CO2 incubator for 3 hr. MTT solution was completely removed after 3 hours. After the MTT solution was completely removed, lOOul / well of DMSO solution was added to each well to dissolve formazan blue-violet crystal (formazan crystal). Absorbance was measured at 595 run. The cell proliferation rate of each sample was calculated based on the blank.

[00081] Biopsy study design: This was a pilot investigation study conducted in accordance with the principles of the 2013 Declaration of Helsinki and approved by an independent institutional review board. The purpose of investigation was to evaluate the histochemical skin changes after a stabilized biomimetic complex use. A total of 13 subjects were enrolled in the study. Informed consent was obtained from all individual participants included in the study. The subjects between the ages of 40 and 55, with Fitzpatrick skin type I-III, in good general health and with mild-to-moderate photo-damaged skin were recruited for the study. Subjects were excluded if they had used topical anti-aging within 30 days of study entry, or topical prescription retinoids within 90 days, or systemic retinoids within 6 months. The subjects were instructed not to use any products throughout the study. Paired tissues from the same individual were treated either by a stabilized- biomimetic complex or by its corresponding vehicle daily by randomization. The subjects used a 0.1 % stabilized biomimetic complex lotion and its corresponding vehicle once daily in the evening for 52 weeks. At end of treatment with the respective formulations, 2-mm punch biopsies were extracted from the subjects. The samples were later shipped to a pathology lab (AIMS Delhi) in 10% buffered neutral formalin for paraffin embedding and for HABP staining process.

[00082] Results and Summary Discussion

[00083] Skin Elasticity Test Name R2 increase % Blank -4.09 CG 1 0.53 CG2 0.29 CG3 0.79 CG4 0.92 CG5 0.92 CG6 1.42 CG7 2.84 CG8 1.89 EG 1 4.34 EG 2 3.3 EG 3 4.3

[00084] Fatigue Resistance (F4) Test Name F4 decrement (%) Blank 3.97 CG 1 3.47 CG2 4.46 CG3 11.8 CG4 11.46 CG5 15.76 CG6 15.35 CG7 13.73 CG8 10.21 EG 1 18.94 EG 2 17.95 EG 3 20.33

[00085] The CK probe is used for evaluating the skin elasticity and the fatigue resistance, the skin careful composition has obvious advantages in the aspects of timely improving the skin tightness and the fatigue resistance, the skin elasticity can be improved by 3.3-4.34%by smearing a sample for30min, and the skin fatigue resistance is reduced by 17.95-20.33%.

[00086] Results of cell counting experiments Name Cell Growth (%) CG 1 20.7 CG2 21.31 CG3 21.75 CG4 23.81 CG5 38.24 CG6 43.4 CG7 45.17 CG8 32.42 EG 1 58.99 EG 2 48.28 EG 3 58.88

[00087] MTT staining test results Name Cell proliferation Rate (%) CG 1 17.76 CG2 16.82 CG3 17.76 CG4 23.36 CG5 36.45 CG6 42.99 CG7 48.6 CG8 36.17 EG 1 64.49 EG 2 50.47 EG 3 63.55

[00088] The skin refining composition is applied to fibroblasts, and through cell counting and MTT chromosome evaluation, the skin refining composition provided by the invention has the advantages that the number of the fibroblasts can be remarkably increased to 48.28-58.88%, and the proliferation rate of the fibroblasts is 50.47% -64.49%.

[00089] Topical stabilized Biomimetic Anti-Aging Treatment induces the expression in human skin

[00090] To further confirm the effect of a stabilized Biomimetic Complex on HA production, we performed studies on full-thickness human skin explants. The HABP staining of three representatives’ images of the vehicle-treated (VEH) human skin explant tissues and the 0.04% stabilized-biomimetic complex (BMC) -treated skin explant tissues were shown in below figure. The HABP staining was in a diffuse pattern in the extracellular space between the epidermal keratinocytes and the dermal fibroblasts as shown in purple color with no nuclear counter-staining. Topical application of the stabilized biomimetic complex at 0.04% for 7 days showed darker purple color of HABP staining in epidermis and in dermis of the skin explants than that of the vehicle-treated skin explants, suggesting a stabilized-biomimetic complex formulation enhanced HA production in human skin explants.

[00091] Paired tissues from each individual were treated either by the vehicle or a stabilized-BMC formulation daily for 52 weeks. The HABP staining of representative epidermal images of three pairs of the vehicle-treated tissues and the stabilized-BMC-treated tissues was shown in figure below. The HABP staining was in a diffuse pattern in the extracellular space between the epidermal keratinocytes as shown in blue color. Topical application of a stabilized-BMC (0.1%)-treated tissues showed darker blue color of HABP staining in the extracellular space of the viable layers of the epidermis as indicated by arrows (—>), which suggested that the stabilized-BMC formulation induced the production of HA in the extracellular space of the viable layers of the epidermis as compared to that of the vehicle-treated tissues figure below. The HABP-staining intensities of the paired tissue of the vehicle-and the stabilized-BMC treatments were quantitated, as shown in figure below. The HABP staining of the paired tissues of the stabilized-BMC treatment was significantly higher than that of the vehicle treatment (p = 0.0076). These histological results demonstrate that a stabilized retinol formulation can continue to induce biological responses and sustain increased levels of HA in the epidermis in vivo over a year of continuous use. The epidermal thickness of the treated tissues was also quantitated. To confirm the efficacy of the stabilized BMC, the epidermis thickness of the treated samples was measured. The epidermis thickness of the stabilized-BMC-treated tissue is 117± 5 pm, whereas the epidermis thickness of the vehicle treated tissue is 83 ± 4 pm. The stabilized- BMC treatment increased the epidermal thickness as compared to that of the vehicle-treated tissues figure below.

[00092] FIG7. shows that the present Biomimetic Anti- Aging Cream employs a barrier lesion on 3D skin model.

[00093] Thus, the proposed products will be a breakthrough in technology and formulation of natural skin care products using biomimetics (by using such natural ingredients that mimic to those actually existing in the skin for improving its appearance and immunity) that will help the skin in improving its self-defense mechanism and capacity to maintain its heath rather than getting support form external synthetic materials causing harm in the long run. The proposed formulations only express several combinations of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

[00094] By imitating the skin's natural composition, it provides the skin with only high tolerance, 100% natural-origin ingredients. Biomimetic skincare incorporates ingredients naturally found in skin such as water, lipids, proteins, sugars and mineral salts. Your skin is therefore naturally in harmony with these skincare products, it “accepts” them and knows how to use them to help replenish the skin's natural barrier. Biomimetic skin care is an advanced approach to formulation utilising innovative plant-based and synthetic biomimetic ingredients that integrate nature and science. These ingredients mimic skin structures and biochemicals which enable optimal delivery and results; this is the next frontier in natural skin care. “Biomimetic materials [are] triggers for our own natural production of collagen, elastin, sebum, etc., instead of applying materials like collagen on skin, which is useless.” Biomimicry has been the catalyst for the discovery of powerful anti-aging peptides, how to reignite slowed collagen production, and superior moisturizing ingredients. Given the growing market demand for organic products and the emerging trend of biomimetics particularly in the skin care industry offers a progressive scope of skin products that improves the health of skin giving it a long-lasting natural impact. Product Composition for Biomimetics Anti-ageing Cream and Serum The cream and serum are designed to balance the skin microbiome, reduce age spots and discoloration, and provide ultra-hydration and ultra-brightening using biomimetics. The proposed unique products contain prebiotic, postbiotic, and probiotic ingredients in such a unique and optimized ratio that it balances and nourish the skin microbiome (trillions of microorganisms, like bacteria, fungi, and viruses found on the skin as invisible lifeforms are known as the skin microbiome). Our Optimized mix of natural prebiotics, probiotics, and postbiotics ingredients derived from natural processes such as fermentation and other organic sources that balance and nourish the skin microbiome. The unique antiaging cream contains prebiotic, postbiotic, and probiotic ingredients that balance and nourish the skin microbiome. Overall, our biomimetic anti-aging cream with prebiotic, postbiotic, and probiotic ingredients is a lab proven solution for those looking for a vegan and cruelty-free way to achieve youthful, healthy-looking skin. The unique biomimetic antiaging serum contains prebiotic, postbiotic, and probiotic ingredients that balance and nourish the skin microbiome. Overall, our biomimetic anti-aging serum with prebiotic, postbiotic, and probiotic ingredients is a lab proven solution forthose looking for a vegan and cruelty-free way to achieve youthful, healthy-looking skin.

[00095] The foregoing descriptions of specific embodiments of the present disclosure have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the present disclosure to the precise forms disclosed, and many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described to best explain the principles of the present disclosure and its practical application, and to thereby enable others skilled in the art to best utilize the present disclosure and various embodiments with various modifications as are suited to the particular use contemplated. It is understood that various omissions and substitutions of equivalents are contemplated as circumstances may suggest or render expedient, but such omissions and substitutions are intended to cover the application or implementation without departing from the scope of the present disclosure.

Claims

1. A biomimetic anti-aging skincare composition comprising:a liposomal blend having a mixture of:(i) one or more prebiotics,(ii) one or more probiotics, and(iii) one or more postbiotics;a gel network comprising water and a gellant; andone or more cosmetically acceptable additives selected from emollients, humectants, peptides, ceramides, vitamins, antioxidants, emulsifiers, preservatives, plant extracts or a combination thereof, wherein the composition is adjusted at a pH in the range of 4.5 to 5.5 to mimic the natural acid mantle of human skin.

2. The composition of claim 1, wherein the one or more prebiotics are selected from oat meal extract, banana extract, asparagus extract or a combination thereof in a range of 0.6 % to 1 % by weight of the total weight of the liposomal blend.

3. The composition of claim 1, wherein the one or more probiotics are selected from Lactobacillus plantarum (KPBBC7), Lactobacillus Arizonensis Ferment Filtrate, Bacillus Ferment &Saccharomyces Ferment Filtrate, Kombucha Tea, Lactobacillus Ferment Lysate, Bacillus Ferment &Saccharomyces Ferment Filtrate or a combination thereof in a range of 0.5 % to 5 % by weight of the total weight of the liposomal blend.

4. The composition of claim 1, wherein the one or more postbiotics are selected from alpha-glucan oligosaccharide, metabolites of Lactobacillus delbrueckii or L. fermentum, or bacterial fermentation supernatant or a combination thereof in a range of 1 % to 2.5 % by weight of the total weight of the liposomal blend.

5. The composition of claim 1, wherein the liposome blend are phospholipid-based vesicles formed using thin-film hydration followed by sonication or extrusion to achieve a particle size ranging from 50 to 200 nm.

6. The composition of claim 1, wherein the antioxidant comprises Undaria pinnatifida cell culture extract in an amount ranging from 0.5% to 1.5% by weight.

7. A method of preparing a biomimetic anti-aging skincare composition, the method comprising:heating a water phase comprising humectants and water-soluble ingredients ata temperature in a range of 70°C to 80°C;heating an oil phase comprising carrier oils, emulsifiers, and oil-soluble ingredients at a temperature in a range of 70°C to 80°C;combining the oil phase with the water phase under high-shear stirring to form an emulsion;cooling the emulsion at a temperature in a range of 30°C to 40°C;adding a liposomal blend having one or more prebiotics, one or more probiotics, and one or more postbiotics to the emulsion under gentle mixing;adjusting the pH of the final composition to between 4.5 and 5.5; and optionally adding fragrances, preservatives, antioxidants, and additional actives and homogenizing the composition to obtain a stable anti-aging skincare composition.

8. The method of claim 7, wherein preparing the liposomal blend comprising: dissolving phospholipids and cholesterol in an organic solvent; evaporating the organic solvent to form a lipid film;hydrating the lipid film with an aqueous solution containing the one or more prebiotics, the one or more probiotics, and the one or more postbiotics; and sonicating the lipid film to a desired nanosized liposomal blend.

9. The method of claim 8, wherein the desired nanosized liposomal blend has a particle size distribution in the range of 50 to 200 nm.

10. The method of claim 7, wherein the composition is homogenized post-addition of the liposomal blend using a high-shear mixer at a speed of 1,000-3,000 rpm for 5-10 minutes to ensure uniform distribution.

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