Topical skin care composition for modulating epidermal gene expression

Abstract

A topical skincare composition is provided, made from a mixture of hydroxytyrosol, green coffee extract, and green tea leaf extract, for modulating epidermal cell gene expression profile to relieve various skin conditions and significantly reduce associated disease symptoms, and to slow the rate of skin aging. More particularly, the topical skincare composition comprises hydroxytyrosol at a concentration of 0.1% w / w to 0.75% w / w, green tea extract at a concentration of 0.025% w / w to 0.1% w / w, and green coffee bean extract at a concentration of 0.025% w / w to less than 0.2% w / w. Also described is a method of treating one or more of signs of aging, skin degradation, collagen, and elastin in a single dose pharmaceutical composition, and a method of activating skin cells having the step of administering the topical skincare composition.

Description

Cross-references to related applications

[0001] This application claims priority to U.S. Provisional Application No. 63 / 706,608, filed October 11, 2024, which is incorporated herein by reference in its entirety. Technical Field

[0002] This disclosure generally relates to skin care products, and more specifically, to a topical skin care composition that alleviates symptoms of various skin conditions by regulating cellular gene activity. Background Technology

[0003] Skin care and ensuring a healthy epidermis are key components of overall personal health. As the body's largest organ, the skin protects individuals from external factors such as ultraviolet radiation, oxidative stress, viral infections, bacteria, and other factors that can directly affect normal skin function / homeostasis. Exposure to these factors accelerates changes in cell function and reproductive capacity over time (a condition known as "premature aging"). Skin aging is also an important part of skin health. As skin cells age, they lose their ability to proliferate until they enter an irreversible state of cell cycle arrest (a state known as "senescence").

[0004] Aging is defined as a process in which the function and reproductive capacity of cells within an organ or organism undergo continuous changes over time. Aging is directly associated with increased morbidity and mortality. Within an organism, organs can age at varying rates, which depend on and are directly influenced by exposure to various environmental stressors. Environmental stressors significantly accelerate the aging rate, which is directly related to the type, severity, and duration of exposure to the stressor. It is not uncommon for an organism to exhibit significant differences in organ aging rates (the accelerated changes in cells over time).

[0005] Skin aging is caused by twelve aging markers (“aging markers”). Each marker affects the skin in a specific way, contributing to the signs of aging. The first marker is loss of protein homeostasis. Aging disrupts the balance of protein synthesis, folding, and degradation, leading to the accumulation of dysfunctional proteins in cells. This accumulation of dysfunctional proteins overloads the system and limits the reach of active proteins to their targets. The second marker is cellular senescence. Senescent cells stop dividing and begin secreting inflammatory factors, a major contributor to tissue aging. The third marker is genomic instability. The accumulation of DNA damage leads to genomic instability, resulting in improper protein synthesis or dysfunction, thus accelerating aging. The fourth marker is telomere loss. Shortened telomeres reduce regenerative capacity, contributing to cellular senescence. The fifth marker is epigenetic alterations. Changes in gene expression regulation contribute to aging. Overexpression or underexpression of key protein factors can adversely affect cells, leading to cellular senescence. The sixth marker is mitochondrial dysfunction. Damaged mitochondria reduce energy production and increase oxidative stress, both of which contribute to cellular senescence. The seventh hallmark is nutrient sensing dysfunction. Aging affects the pathways by which nutrients are sensed and responded to, thus disrupting metabolism. The eighth hallmark of aging is stem cell depletion. Depletion of stem cell reserves reduces tissue regeneration, leading to cellular senescence. The ninth hallmark is altered intercellular communication. Aging disrupts cell signaling, leading to tissue dysfunction and cellular senescence. The tenth hallmark of aging is chronic inflammation. Persistent low-grade inflammation accelerates aging. The eleventh hallmark is oxidative stress caused by reactive oxygen species (“ROS”). Excessive ROS damage cellular components and promote aging. The twelfth hallmark is impaired autophagy. Impaired autophagy hinders the clearance of damaged cellular components, which can lead to systemic failure and cellular senescence.

[0006] Similar to and often interrelated with skin aging, skin diseases are typically associated with changes in cellular activity caused by external factors or alterations in cellular pathways. The impact of skin diseases can be devastating for an individual, as the skin is an outward manifestation of personal health. The skin possesses numerous maintenance factors that prevent or slow disease progression. These maintenance factors are regulated through cellular sensing or signaling pathways. A topical skincare composition that can influence these internal factors and upregulate their effects, thereby enhancing its ability to prevent or slow disease progression, would be advantageous.

[0007] Proper skin care is achieved through innate / adaptive mechanisms (“cellular responses”) and behavioral adjustments (“behavioral responses”). Cellular responses include: (i) the skin’s own mechanisms for maintaining normal function, and (ii) cellular responses to changes or external stresses within the dermis or in the environment. Behavioral responses include an individual’s behaviors in maintaining their own skin.

[0008] Normal skin function is crucial to an individual's health. As the body's first point of contact with the external environment, the skin adjusts its function to ensure it can adapt to and overcome external stresses and various attacks. As an organ, the skin possesses multiple mechanisms to maintain its normal homeostasis and health—namely, innate and adaptive mechanisms. If these mechanisms are defective, malfunction, or bypassed by various factors or mutations, then the skin as an organ will break down, and various diseases will manifest and show symptoms on the skin's surface.

[0009] The skin possesses a variety of adaptive mechanisms (“defense strategies”) to effectively protect the skin and internal organs from a range of external factors. These defense strategies are partly mediated by the gene expression profiles of individual skin cells (“epidermal cells”). Gene expression profiles within the skin are regulated by the influence of various external factors, the accumulation of active substances, or the activation of upstream regulatory proteins. In the simplest case, due to overexposure to a stressor (ultraviolet radiation from the sun), epidermal cells increase the expression of genes responsible for melanin production. Following expression, melanin absorbs harmful long-wave ultraviolet (UVA), medium-wave (UVB), and short-wave (UVC) light. If ultraviolet radiation is not absorbed due to insufficient melanin expression, it can damage cellular DNA, potentially leading to gene mutations and the development of skin cancer (“melanoma”).

[0010] In some individuals, defense strategies are highly weakened (“affected state”). This affected state can be (but is not limited to) caused by genetic makeup, expression profile, mutations, and age. The affected state can have serious consequences for individuals because the skin may be highly susceptible to various disease manifestations, skin barrier disruption, and the potential onset of melanoma. Individuals in the affected state should adopt behavioral mechanisms to reduce the likelihood of disease manifestations and melanoma onset.

[0011] Behavioral mechanisms are actions individuals can take to further enhance their skin's ability to withstand external stress and aggression. By adjusting their daily routines, individuals can reduce the amount and severity of external stress on their skin, thus maintaining normal skin homeostasis and health without activating the skin's defense strategies. Some behavioral mechanisms include: marketed reductions in harmful UV exposure, use of antibacterial ointments, use of sunscreen, and use of skincare products containing various ingredients ("skincare elements").

[0012] Many skincare products contain skincare elements that have previously been proven effective in treating skin conditions and alleviating symptoms associated with skin diseases. These skincare elements are only beneficial when they are provided in their active, bioavailable form and targeted to the epidermal cells where they can function. For example, collagen is a commonly used skincare element in various products. The collagen provided in most skincare products does not target the cytoplasm of epidermal cells but remains on the skin's surface. At this location, its effects may be minimal because epidermal cells cannot utilize collagen in the way required for normal cellular function—to create and subsequently secrete type I and type II collagen fibers. Furthermore, using collagen as a product ingredient does not activate cellular mechanisms for collagen transcription and subsequent regulation.

[0013] The three adaptogenic ingredients already used in skincare products are: hydroxytyrosol; green tea extract; and green coffee extract (collectively referred to as "product ingredients"). First, hydroxytyrosol is a phenolic compound found in the fruit and leaves of the olive (Olea europaea), most commonly obtained through the hydrolysis of oleuropein. Hydroxytyrosol is one of the main components of extra virgin olive oil, olive processing wastewater, and olive leaf extract; its chemical formula is C8H₂O. 10 O3. Hydroxytyrosol has been shown to be a phytochemical with antioxidant properties, as well as anti-inflammatory, antiviral, antibacterial, and antifungal properties. Numerous patent applications describe the benefits of using hydroxytyrosol in combination with other elements. One specific benefit of the use of hydroxytyrosol relates to its beneficial application as an antioxidant in the treatment of oxygen free radical-related diseases. US Patent Application US 2013-0266545 describes a composition and method for wound healing and inhibition of necrosis.

[0014] Secondly, green tea extract is a naturally occurring antioxidant. It is an effective way to promote collagen production in the skin. Green tea extract has also been shown to increase skin firmness while improving redness, elasticity, and overall texture. Furthermore, green tea extract is an excellent moisturizer.

[0015] Finally, green coffee extract is a naturally occurring antioxidant. It's an effective way to promote collagen production in the skin. Green coffee bean extract has also been shown to increase skin firmness while improving redness and overall texture.

[0016] While it's generally believed that using skincare products containing antioxidants will help reduce free radicals, they haven't previously been shown to act as effective activators of cellular transcription factors, thereby inducing alterations in epidermal cell gene expression. Antioxidants have only shown their potential to reduce free radicals within skin cells, which can lead to DNA damage if left untreated. Reducing free radicals may decrease the likelihood of DNA damage (caused by free radicals), but it doesn't affect cellular defense strategies associated with repairing any damage already caused by free radicals. Furthermore, it hasn't previously been shown that using such skincare elements can help affected cells increase the generation of defense strategies.

[0017] Various attempts have been made to describe the effects of product ingredients on the skin. Prior publications such as U.S. Patent Nos. 9,511,034 (Garrett); 11,266,594 (Florence); and 9,889,171 (Chevreau) provide examples of using product ingredients to manage and treat the skin.

[0018] Garrett discloses a skin treatment system containing hydroxytyrosol for treating a variety of skin conditions, ranging from scars (including keloids) to burns and wrinkles. Specifically, Garrett discloses an elastomeric component containing organosilicon to promote wound healing and shorten curing time. Garrett has not considered using this compound to modulate DNA repair mechanisms, nor has it considered using it to prevent or combat melanoma, or to increase the transcription of various genes necessary for optimal cell and skin function, as well as for adjuvant defense strategies or reversing skin aging. Furthermore, Garrett has not considered using green tea or green coffee beans.

[0019] Florence discloses a topical skincare formulation comprising plant extracts for treating skin conditions caused by exposure to pollution or free radicals. Florence discloses a formulation containing an effective amount of African persimmon leaf extract. The disclosed skincare formulations aim to reduce oxidative damage in the skin. Florence does not describe the use of hydroxytyrosol, green tea extract, and green coffee extract for the prevention or treatment of melanoma. Furthermore, Florence does not disclose topical skincare formulations for increasing collagen synthesis, regulating extracellular matrix (ECM) remodeling, or modulating cellular defense strategies through gene expression profiling.

[0020] Chevreau discloses a topical composition and method for reducing oxidative stress using Brassica plant extracts. Chevreau discloses the use of Brassica plant extracts and Camellia oleifera extract to improve skin appearance and overall health. Topical application of this composition has been shown to mimic the target gene of nuclear factor erythroid 2-related factor 2 (Nfr2) to alleviate oxidative stress. Specifically, catalase expression has been shown to be highly expressed, a key antioxidant enzyme in the body's defense against oxidative stress. Skin appearance is also improved, partly due to increased density of the collagen network in the dermis. While the prior patent does describe a topical skin cream containing plant extracts that activate catalase gene expression to reduce oxidative stress and increase collagen formation, it fails to use hydroxytyrosol, green coffee extract, or green tea extract, and fails to activate multiple gene pathways crucial for skin regeneration, nor does it describe the use of the compound in the prevention or fight of melanoma.

[0021] Therefore, there is a need for a topical skincare composition that activates cellular transcription factors in multiple modes to increase gene transcription and subsequent cellular protein production. This includes a topical skincare composition capable of increasing gene transcription factors to improve and strengthen cellular defense mechanisms, increasing DNA repair molecules and upregulating upstream factors, and increasing the production of effective cytokines crucial for cellular health and reversing or reducing skin aging. It also includes a skincare composition capable of modulating gene expression to activate cellular repair mechanisms associated with DNA damage and activating DNA expression profiles crucial for rebuilding cellular components. This modulation should have a dual effect: firstly, alleviating the symptoms of skin conditions or infections; and secondly, reducing their likelihood of occurrence. Indeed, there is a need for a topical skincare composition that can be used in various skincare products to reduce the likelihood of various skin conditions and, if present, significantly alleviate their associated symptoms, thereby restoring the skin to its natural healthy homeostasis. Summary of the Invention

[0022] This disclosure provides a topical skin composition for alleviating or eliminating various skin diseases. This topical skin composition rebuilds and strengthens key cellular components and restores skin health and homeostasis by modulating epidermal cell gene expression profiles and activating cellular repair mechanisms. Thus, this topical skin composition achieves two main functions related to skin treatment: 1. reducing or eliminating the likelihood of skin diseases; 2. alleviating the severity of existing skin conditions. Modulating epidermal cell gene expression profiles enables cells to regulate defense strategies through the modulation of innate and adaptive mechanisms. Furthermore, this topical skin composition provides skin cells with a mechanism to reverse signs of aging and normalize / help the epidermis by: reducing reactive oxygen species (ROS) under stress; reducing interleukin-6 (IL-6); increasing and stabilizing hypoxia-inducible factor-1 alpha (HIF-1α); increasing the production of DNA damage-induced transcription factor 4 (DDIT4); regulating angiogenesis and apoptosis factors; increasing collagen synthesis; and regulating extracellular matrix (ECM) remodeling.

[0023] This topical skin composition can be formulated into topical skin care products (moisturizers, sunscreens, serums) to protect the skin from various skin conditions, or to treat and alleviate the symptoms or severity of existing skin conditions.

[0024] In one aspect, this disclosure provides a method for activating skin cells, the method comprising: providing a topical skin composition comprising: 0.1% w / w (weight / weight) to 0.75% w / w hydroxytyrosol; 0.025% w / w to 0.1% w / w green tea extract; and 0.025% w / w to less than 0.2% w / w green coffee bean extract; and applying the topical skin composition topically to the skin.

[0025] In another aspect, this disclosure provides a topical skin composition comprising: 0.1% w / w to 0.75% w / w hydroxytyrosol; 0.025% w / w to 0.1% w / w green tea extract; and 0.025% w / w to less than 0.2% w / w green coffee bean extract; wherein the topical skin composition is applied to the surface of epidermal cells to modulate cellular gene expression profiles that are essential for skin health, prevention of skin diseases, and reduction of the severity of affected skin cells.

[0026] In another aspect, this disclosure provides a method for treating one or more of signs of aging, skin degeneration, collagen, and elastin in the form of a single-dose pharmaceutical composition, the method comprising: providing a pharmaceutical composition comprising: 0.1% w / w to 0.75% w / w hydroxytyrosol; 0.025% w / w to 0.1% w / w green tea extract; and 0.025% w / w to less than 0.2% w / w green coffee bean extract; and applying the pharmaceutical composition topically to the skin. Attached Figure Description

[0027] For convenience, this disclosure will now be described in conjunction with a specific embodiment of this disclosure. It should be understood that the accompanying drawings only relate to one embodiment of this disclosure and are not intended to limit this disclosure.

[0028] Figure 1 shows the quantitative polymerase chain reaction (qPCR) analysis of prolyl 4-hydroxylase 1 messenger RNA (P4H1 mRNA) expressed in epidermal cells. Prolyl 4-hydroxylase 1 messenger RNA is associated with collagen synthesis. Epidermal cells were incubated with various creams or cream carriers and cultured with or without UV radiation. Epidermal cells were tested using Protectif Tinted and Renutriate Professional, and their respective messenger RNA (mRNA) expression was compared with that of epidermal cells treated with a vector (the vector being Renutriate Professional and Protectif Tinted, which do not contain the ingredients of this product). Cells treated with Renutriate Professional and Protectif Tinted showed a more than three-fold increase in prolyl 4-hydroxylase subunit α1 messenger RNA (P4H1 mRNA).

[0029] Figure 2 shows the quantitative polymerase chain reaction (qPCR) analysis of LAMB1 messenger RNA (mRNA) expressed in epidermal cells. LAMB1 messenger RNA is associated with extracellular matrix (ECM) remodeling. Epidermal cells were incubated with various creams or cream carriers, with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective mRNA expression was compared with epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the product's ingredients). Cells treated with Renutriate Professional and Protectif Tinted products can increase the amount of Lamin B1 messenger RNA (LAMB1 mRNA) by more than 1.5 times.

[0030] Figure 3 shows the quantitative polymerase chain reaction (qPCR) analysis of LAMB4 mRNA (Lamin B4 messenger RNA) expressed in epidermal cells. LAMB4 mRNA is associated with extracellular matrix (ECM) remodeling. Epidermal cells were incubated with various creams or cream carriers, with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective mRNA expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted without the product's ingredients). Cells treated with Renutriate Professional and ProtectifTinted products can increase the amount of Lamin B4 messenger RNA (LAMB4 mRNA) by more than 1.5 times.

[0031] Figure 4 shows the quantitative polymerase chain reaction (qPCR) analysis of LAMA4 mRNA (Lamin Amessenger RNA) expressed in epidermal cells. LABA4 mRNA is associated with extracellular matrix (ECM) remodeling. Epidermal cells were incubated with various creams or cream carriers, with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective mRNA expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted without the product's ingredients). Cells treated with Renutriate Professional and ProtectifTinted products can increase the amount of Lamin A4 messenger RNA (LAMA4 mRNA) by more than 1.5 times.

[0032] Figure 5 shows the quantitative polymerase chain reaction (qPCR) analysis of decorin messenger RNA (DCN mRNA) of core proteoglycan expressed in epidermal cells. DCN mRNA is associated with extracellular matrix (ECM) remodeling. Epidermal cells were incubated with various creams or cream carriers, with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective messenger RNA (mRNA) expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the ingredients of this product). Cells treated with Renutriate Professional and ProtectifTinted products can increase the amount of decorin messenger RNA (DCN mRNA) in the core proteoglycan by more than 1.5 times.

[0033] Figure 6a shows a quantitative polymerase chain reaction (qPCR) analysis of interleukin-6 messenger RNA (IL6 mRNA) expressed in epidermal cells. IL6 mRNA is associated with stress and inflammatory responses. Epidermal cells were incubated with various creams or cream carriers in the presence or absence of UV radiation. Skin cells were tested with Protectif Tinted and Renutriate Professional, and their respective mRNA expression was compared with epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain any of the product's ingredients). Cells treated with Renutriate Professional and Protectif Tinted products can reduce interleukin-6 messenger RNA (IL6 mRNA) by more than two times.

[0034] Figure 6b is a quantitative protein analysis of interleukin-6 (IL6) protein expressed in epidermal cells. Interleukin-6 (IL6) protein is associated with stress and inflammatory responses. Epidermal cells were incubated with various creams or cream carriers in the presence or absence of UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective protein expression was compared with epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted without the product's ingredients). Cells treated with Renutriate Professional and Protectif Tinted showed a 125 pg / ml reduction in interleukin-6 (IL6) production compared to the corresponding carriers.

[0035] Figure 7a shows the quantitative polymerase chain reaction (qPCR) analysis of interleukin-1β messenger RNA (IL1b mRNA) expressed in epidermal cells. Interleukin-1β messenger RNA (IL1b mRNA) is associated with stress and inflammatory responses. Epidermal cells were incubated with various creams or cream carriers in the presence or absence of UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective mRNA expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted without the product's ingredients). Cells treated with Renutriate Professional and Protectif Tinted products can reduce interleukin-1β messenger RNA (IL1b mRNA) by more than three times.

[0036] Figure 7b is a graph showing the quantitative analysis of interleukin-1β (IL1b) protein expressed in epidermal cells. Interleukin-1β protein is associated with stress and inflammatory responses. Epidermal cells were incubated with various creams or cream carriers and cultured with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective protein expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted without the product's ingredients). Cells treated with Renutriate Professional and Protectif Tinted showed a 60 pg / ml reduction in interleukin-1β (IL1b) production compared to the corresponding carriers.

[0037] Figure 8 shows the quantitative polymerase chain reaction (qPCR) analysis of DNA damage-induced transcription factor 4 (DDIT4) messenger RNA expressed in epidermal cells. DDIT4 mRNA is associated with stress and inflammatory responses. Epidermal cells were incubated with various creams or cream carriers in the presence or absence of UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective messenger RNA (mRNA) expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the ingredients of this product). Cells treated with Renutriate Professional and Protectif Tinted products can increase DNA damage-induced transcription factor 4 (DDIT4) messenger RNA by more than 1.5 times.

[0038] Figure 9 shows the quantitative polymerase chain reaction (qPCR) analysis of VEGF mRNA (vascular endothelial growth factor messenger RNA) expressed in epidermal cells. VEGF mRNA is associated with angiogenesis. Epidermal cells were incubated with various creams or cream carriers, with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective messenger RNA (mRNA) expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the ingredients of this product). Cells treated with professional repair and nourishing products and protective tints can more than double the amount of vascular endothelial growth factor (VEGF) messenger RNA.

[0039] Figure 10 shows the quantitative polymerase chain reaction (qPCR) analysis of B-cell lymphoma-2 (BCL2) interacting protein 3 messenger RNA (BNIP3 mRNA) expressed in epidermal cells. BNIP3 mRNA is associated with apoptosis. Epidermal cells were incubated with various creams or cream carriers in the presence or absence of UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective messenger RNA (mRNA) expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the ingredients of this product). Cells treated with Renutriate Professional and ProtectifTinted products can reduce the amount of B-cell lymphoma-2 (BCL2) interacting protein 3 messenger RNA (BNIP3 mRNA) by more than three times.

[0040] Figure 11 is a quantitative analysis of the protein expression of hypoxia-inducible factor-1α (HIF-1α) in epidermal cells. HIF-1α protein is associated with cellular stress and the presence of oxidative reactive substances. Epidermal cells were incubated with various creams or cream carriers under conditions with or without UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective protein expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the product's ingredients). Cells treated with Renutriate Professional and Protectif Tinted were able to produce 50 pg / ml more hypoxia-inducible factor-1α (HIF-1α) protein than their respective carriers.

[0041] Figure 12 is a quantitative analysis of reactive oxygen species (ROS) production in epidermal cells. ROS are associated with oxidative stress, which can lead to cell damage (lipid peroxidation and DNA fragmentation). Epidermal cells were incubated with various creams or cream carriers in the presence or absence of UV radiation. Epidermal cells were tested with Protectif Tinted and Renutriate Professional, and their respective protein expression was compared with that of epidermal cells treated with carriers (Renutriate Professional and Protectif Tinted, which do not contain the ingredients of this product). Cells treated with Renutriate Professional and Protectif Tinted showed a 200-unit reduction in normalized ROS levels compared to their respective carriers in the presence of UV radiation. As expected, in the absence of ultraviolet light, reactive oxygen species (ROS) did not increase.

[0042] Figures 13A to 13E This is an image of the patient's (Patient 1) head while using Renutriate PRO. Patient 1 used a small amount of Renutriate PRO twice daily for eight months.

[0043] Figure 13A This is the patient's skin on day 0.

[0044] Figure 13B This refers to the skin of patients who have been continuously using the professional repair and nourishing product (Renutriate PRO) for up to 60 days. Figure 13C This refers to the skin of patients who have been using the professional repair and nourishing product (Renutriate PRO) for 120 days.

[0045] Figure 13D This refers to the skin of patients who have continuously used the professional repair and nourishing product (Renutriate PRO) for 180 days.

[0046] Figure 13E After 240 days of continuous use of the professional repair and nourishing product (Renutriate PRO), the patient's skin showed a revitalized and healthy appearance.

[0047] Figure 14A This is a picture of the patient's skin on day 0 after 240 days of continuous use of Renutriate PRO. Figure 14 is an image of the patient's face during the use of Renutriate PRO. The patient used a small amount of Renutriate PRO twice daily for four months.

[0048] Figure 14B This refers to the skin of patients who have been continuously using the professional repair and nourishing product (Renutriate PRO) for up to 60 days.

[0049] Figure 14C This refers to the skin of patients who have been using the professional repair and nourishing product (Renutriate PRO) for 120 days.

[0050] Figures 15A to 15C These are images of Patient 3's face during the use of Renutriate PRO product. Patient 3 used a small amount of Renutriate PRO twice daily for four months.

[0051] Figure 15A This is the patient's skin on day 0.

[0052] Figure 15B This refers to the skin of patients who have been continuously using the professional repair and nourishing product (Renutriate PRO) for up to 60 days.

[0053] Figure 15C This refers to the skin of patients who have been using the professional repair and nourishing product (Renutriate PRO) for 120 days.

[0054] Figure 16A and 16B These are images of Patient 4's face during the use of Renutriate PRO. Patient 4 used a small amount of Renutriate PRO twice daily for two months.

[0055] Figure 16A This is the patient's skin on day 0.

[0056] Figure 16B This refers to the skin of patients who have been continuously using the professional repair and nourishing product (Renutriate PRO) for up to 60 days.

[0057] Figure 17A and 17B These are images of Patient 5's legs during the use of Renutriate PRO product. Patient 5 used a small amount of Renutriate PRO twice daily for two months.

[0058] Figure 17A This is the patient's skin on day 0.

[0059] Figure 17B This refers to the skin of patients who have been continuously using the professional repair and nourishing product (Renutriate PRO) for up to 60 days.

[0060] Figure 18A and 18B This image shows Patient 6's shoulder during the use of Renutriate PRO product, applied only to the left shoulder. Patient 6 used a standard over-the-counter moisturizer on the right shoulder. Patient 6 applied a small amount of Renutriate PRO twice daily to the left shoulder for two weeks (14 days) after experiencing a minor sunburn.

[0061] Figure 18A This image shows the patient's left shoulder two weeks (14 days) after using a professional repairing and nourishing product (Renutriate PRO) following a minor sunburn.

[0062] Figure 18BThis is an image of the patient's right shoulder two weeks (14 days) after using a standard over-the-counter moisturizer following a minor sunburn.

[0063] Figures 19A to 19C These are images of Patient 7's face during the use of the Renutriate product. Patient 7 used a small amount of Renutriate twice daily for sixteen weeks.

[0064] Figure 19A This is the patient's skin on day 0.

[0065] Figure 19B This is the patient's skin after eight weeks of continuous use of the Renutriate product.

[0066] Figure 19C This is the skin of a patient after sixteen weeks of continuous use of the Renutriate product.

[0067] Figures 20A to 20C This is an image of Patient 8's hand while using the Renutriate product. Patient 8 used a small amount of Renutriate twice daily for two weeks.

[0068] Figure 20A This is the patient's skin on day 0.

[0069] Figure 20B This is the patient's skin after one week of continuous use of the repairing and nourishing product (Renutriate).

[0070] Figure 20C This is the patient's skin after two weeks of continuous use of the Renutriate product.

[0071] Figures 21A to 21D This is an image of Patient 9's ankle during the use of the Renutriate product. Patient 9 used a small amount of Renutriate twice daily for sixteen weeks.

[0072] Figure 21A This is the patient's skin on day 0.

[0073] Figure 21B This is the patient's skin after eight weeks of continuous use of the Renutriate product.

[0074] Figure 21C This is the skin of a patient after twelve weeks of continuous use of the Renutriate product.

[0075] Figure 21D This is the skin of a patient after sixteen weeks of continuous use of the Renutriate product.

[0076] Figures 22A to 22C This is an image of Patient 10's hands during the use of the Renutriate product. Patient 10 used a small amount of Renutriate twice daily for sixteen weeks.

[0077] Figure 22A This is the patient's skin on day 0.

[0078] Figure 22B This is the patient's skin after eight weeks of continuous use of the Renutriate product.

[0079] Figure 22C This is the patient's skin after 16 weeks of continuous use of the Renutriate product.

[0080] Figures 23A to 23C These are images of Patient 11's face during the use of the Renutriate product. Patient 11 used a small amount of Renutriate twice daily for sixteen weeks.

[0081] Figure 23A Showing the patient's face on day 0.

[0082] Figure 23B This is the patient's skin after eight weeks of continuous use of the Renutriate product.

[0083] Figure 23C This is the patient's skin after 16 weeks of continuous use of the Renutriate product.

[0084] Figure 24A and 24B These are quantitative analysis and ultrasound images of the skin age of Patient 12's left and right forearms. Patient 12 used Renutriate PRO, a professional repair and nourishing product, once daily on their left forearm for nine months. Patient 12 used a leading brand product, once daily on their right forearm, for nine months.

[0085] Figure 24A This is a skin age assessment of the patient's left forearm at age 12.

[0086] Figure 24B This is a skin age assessment of the patient's right forearm at age 12.

[0087] Figures 25A to 25C This is a quantitative analysis and assessment of the skin age of the patient's 13 arms. The arms were first treated with Renutriate PRO or leading brand products for 20 minutes, followed by UVB irradiation.

[0088] Figure 25A This is a visual representation of the patient's 13 arms at the pre-UVB irradiation treatment site.

[0089] Figure 25B This is a skin age assessment of patient 13's skin treated with Renutriate PRO.

[0090] Figure 25C This is a skin age assessment of patient 13 whose skin has been treated with leading brand products.

[0091] Figure 26A and 26B This is a quantitative analysis and assessment of the skin age of patient 14 90 minutes after applying the professional repair and nourishing product (Renutriate PRO).

[0092] Figure 26A This is a skin age assessment of patient 14's face before applying the professional repair and nourishing product (Renutriate PRO).

[0093] Figure 26B This is a skin age assessment of the patient's face after treatment with the professional repair and nourishing product (Renutriate PRO). Detailed Implementation

[0094] This disclosure will now be described more fully with reference to the accompanying drawings, which illustrate preferred and other embodiments of this disclosure. No embodiment described below limits any claimed disclosure, and any claimed disclosure may cover processes or elements not described below. The claimed disclosure is not limited to products or elements having all the features of any element or process described below, nor is it limited to features common to multiple or all of the elements described below. An element or process described below may not be an embodiment of any claimed disclosure. The applicant, inventor, or owner reserves all rights they may have in any disclosure claimed in this document, such as rights to claim that disclosure in a continuation application, and does not intend to waive, deny, or contribute any such disclosure to the public by means of disclosure in this document.

[0095] A novel topical skin composition for treating skin conditions and various autoimmune diseases has been developed, offering significant advantages over existing drugs or products. Specifically, the topical skin composition of this disclosure has been shown to effectively modulate gene transcription factors of key regulators of various defense strategies in epidermal cells. This allows individual epidermal cells to effectively alleviate disease manifestations and significantly reduce the likelihood of such diseases developing. Furthermore, the regulation of gene expression has been shown to effectively reduce the incidence and severity of various autoimmune diseases. Finally, the regulation of gene expression has been shown to reverse and delay signs of skin aging.

[0096] Overall, this skincare composition aims to alleviate the symptoms of various skin conditions by modulating cellular gene activity. These symptoms have been proven detrimental to skin health. As skin health declines, elasticity is lost, skin conditions become apparent, and skin aging begins. By modulating epidermal cell gene expression and activating cellular repair mechanisms, this topical skin composition significantly increases the total amount of key cellular components, restoring the skin to its natural healthy homeostasis while enhancing its natural repair and defense mechanisms. Restoring the skin to its natural healthy state, while strengthening its defense mechanisms, helps to revitalize the skin, making it healthier and more youthful. This topical skin composition has the following effects on the epidermis: reducing reactive oxygen species (ROS) in the presence of ultraviolet light; reducing interleukin (IL)-6; increasing and stabilizing hypoxia-inducible factor-1α (HIF-1α) through hypoxia-responsive elements; increasing the production of DNA damage-induced transcription factor 4 (DDIT4); regulating angiogenesis and apoptosis factors; increasing collagen synthesis; and regulating extracellular matrix (ECM) remodeling. More specifically, this disclosure relates to a unique cosmeceutical composition that utilizes the synergistic effects of multiple natural compounds to provide enhanced therapeutic effects. This disclosure details the composition, methods of application, and observed clinical results demonstrating its efficacy in dermatological applications.

[0097] The ingredients in this product work together to regulate cellular biological processes, thereby protecting the skin from external factors, autoimmune diseases, and age-related problems. These effects enable the product ingredients to target signs of aging, providing a comprehensive approach to maintaining skin health and restoring it to its natural healthy state. By stimulating collagen production, reducing inflammation, promoting cell repair, and enhancing stress responses, the disclosed product ingredients are specifically formulated to counteract the negative effects of environmental stressors (such as UV exposure), inflammation, and autoimmune diseases. This disclosure describes the use of hydroxytyrosol, but those skilled in the art will understand that hydroxytyrosol is derived from European olive oil, and any derivative thereof can be used as a substitute.

[0098] The product ingredients disclosed herein have been shown to influence the production of proteins that are key cellular components within epidermal cells. This regulation of gene expression has beneficial effects on the skin, including but not limited to protection against various external factors, disease progression, disease severity, and maintenance of skin homeostasis. Gene expression in cells can be observed in one of two ways: messenger RNA (mRNA) expression (quantitative polymerase chain reaction (qPCR) assay); and protein production enzyme-linked immunosorbent assay (ELISA) assay.

[0099] The ingredients of this product can be formulated into various topical skincare products, including moisturizers, sunscreens, and serums. The disclosed product ingredients have been formulated into moisturizer variants currently known as “Renutriate” (Repairing, Advanced, Professional) and sunscreen variants known as “Protectif” (Neutral, Light, Dark) and “Rejuvenat”. Those skilled in the art will understand that the product ingredients can be used or formulated into any other product, including ingestible supplements. To determine the efficacy of the product ingredients, in vitro tests have been conducted on skin cells on the Renutriate Professional variant (“Renutriate PRO”), Renutriate Restorative (“Renutriate”), and Protectif Fair Tint (“Protectif Tint”), and compared with skin cells treated with a carrier of each product (each carrier being the same corresponding product without the product ingredients). The cells were then cultured in the absence of ultraviolet radiation or in the presence of ultraviolet radiation, and their gene and protein expression profiles were subsequently tested.

[0100] As further illustrated in Figure 1, epidermal cells were treated with a cream product or carrier containing the ingredient of this product, and tested with and without UV radiation (stress). Compared to the carrier, under stress (e.g., UV exposure), the ingredient of this product in Renutriate PRO and Protectif Tinted significantly increased the production of collagen messenger RNA (mRNA), as evidenced by the expression of the prolyl 4-hydroxylase subunit α1 (P4H1) gene. Prolyl 4-hydroxylase subunit α1 (P4H1) plays a crucial role in collagen synthesis and is essential for maintaining the structural integrity of the skin. Even in the absence of UV radiation, cells treated with the ingredient of this product produced significantly higher amounts of prolyl 4-hydroxylase subunit α1 (P4H1). This product's ingredients enhance collagen biosynthesis by stimulating prolyl 4-hydroxylase subunit α1 (P4H1), thereby combating collagen loss caused by aging and environmental stress. The increased collagen production induced by this product's ingredients results in firmer, more elastic skin, reduced wrinkles, and improved skin laxity and hollowing around the eyes and face. Furthermore, collagen loss makes skin more fragile and accelerates aging.

[0101] As further illustrated in Figure 2, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested with and without UV radiation (stress). Compared to the carrier, under stress (e.g., UV exposure), the ingredient of this product in Renutriate PRO and Protectif Tinted significantly increased the production of laminin B1 (LAMB1) messenger RNA (mRNA). Laminin B1 (LAMB1) is essential for maintaining nuclear integrity and chromatin organization. Increased expression of laminin B1 (LAMB1) is crucial for achieving optimal epidermal function. Decreased laminin B1 (LAMB1) in cells is associated with cellular senescence, a phenomenon characterized by the cessation of cell division. Cells with low levels of laminin B1 (LAMB1) exhibit permanent proliferative arrest and respond to endogenous and exogenous stresses, including but not limited to telomere dysfunction, oncogene activation, and persistent DNA damage. Cellular senescence is also associated with aging and directly leads to impaired tissue regeneration and age-related chronic diseases. By increasing laminin B1 (LAMB1) expression, the ingredients in this product help maintain genomic stability, reduce cellular dysfunction, and significantly lower the risk of cancer. Furthermore, by increasing laminin B1 (LAMB1) levels in epidermal cells, the ingredients in this product can limit the aging process and maintain a more youthful skin appearance.

[0102] As further illustrated in Figure 3, epidermal cells were treated with a cream product or carrier containing this product ingredient, and tested with and without UV radiation (stress). Compared to the carrier, under stress (e.g., UV exposure), the product ingredient in Renutriate PRO and Protectif Tinted significantly increased the production of laminin B4 (LAMB4) messenger RNA (mRNA). Increased expression of laminin B4 (LAMB4) is crucial for extracellular matrix (ECM) remodeling and enhanced skin elasticity. Laminin B4 (LAMB4) works synergistically with laminin B1 (LAMB1) to support nuclear structure and cell proliferation. This product ingredient enhances extracellular matrix (ECM) remodeling by increasing laminin B4 (LAMB4) expression, maintains skin elasticity, and contributes to a youthful and radiant appearance. One benefit of increasing laminin B4 (LAMB4) is maintaining skin elasticity and preventing sagging. Increased LAMB4 in the epidermis supports the maintenance of nuclear integrity, thus addressing markers of protein homeostasis loss and genomic instability. By increasing nuclear integrity and maintaining skin elasticity, the ingredients in this product reduce signs of aging, leaving skin with a radiant appearance.

[0103] As further illustrated in Figure 4, epidermal cells were treated with a cream product or carrier containing this product ingredient, and tested with and without UV radiation (stress). Compared to the carrier, under stress (e.g., UV exposure), the product ingredient in Renutriate PRO and Protectif Tinted significantly increased the production of laminin A4 (LAMA4) messenger RNA (mRNA). Increased expression of laminin A4 (LAMA4) is crucial for maintaining nuclear structure and effective gene regulation. Laminin A4 (LAMA4) contributes to epidermal nuclear integrity and regulates key gene regulators involved in cell repair and collagen production. By enhancing laminin A4 (LAMA4) expression, this product ingredient strengthens and consolidates the skin's natural repair mechanisms and ensures better resistance to aging. The upregulation of laminin A4 (LAMA4) by this product ingredient functions by responding to epigenetic alterations through appropriate gene expression. The ingredients in this product can maintain proper gene expression and prevent premature cell aging.

[0104] As further illustrated in Figure 5, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested in the presence or absence of ultraviolet radiation (stress). Compared to the carrier, the ingredient of this product in Renutriate PRO and Protectif Tinted significantly increased the production of core proteoglycan (“DCN”) mRNA under both stress-free and stressful conditions (e.g., ultraviolet exposure). Increased expression of core proteoglycan (DCN, decorin messenger) contributes to collagen fiber assembly and extracellular matrix (ECM) stability, while ensuring proper collagen organization through fiber formation. Core proteoglycan (DCN, decorin messenger) is a major proteoglycan in the skin and regulates the assembly of the collagen matrix. Core proteoglycan (DCN, decorin messenger) is an important component of skin connective tissue and is crucial for maintaining the mechanical strength of the skin. This product's ingredients upregulate decorin messenger (DCN) levels, resulting in smoother, firmer skin. DCN prevents extracellular matrix degradation and directly addresses age-related tissue fragility. Degradation or loss of DCN proteins in aging skin (age-dependent changes) leads to skin fragility, common in older adults. DCN also regulates epidermal growth factors such as transforming growth factor-β (TGF-β), further influencing cell growth and differentiation.

[0105] As further illustrated in Figure 6, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested with and without UV radiation (stress). Compared to the carrier, under stress conditions (e.g., UV exposure), the ingredient of this product in Renutriate PRO and Protectif Tinted significantly reduced the production of the key inflammatory marker interleukin-6 (IL-6). Figure 6a assesses the presence of intracellular interleukin-6 messenger RNA (IL-6 mRNA), while Figure 6b assesses the presence of intracellular interleukin-6 protein. The amount of intracellular messenger RNA (mRNA) in epidermal cells correlates with the presence of interleukin-6 protein. Interleukin-6 (IL-6) is a pro-inflammatory cytokine that contributes to chronic inflammation. The anti-inflammatory properties of this product's ingredients are key to preventing long-term skin damage caused by stress. Furthermore, chronic inflammation accelerates aging by promoting tissue breakdown.

[0106] As further illustrated in Figure 7, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested with and without ultraviolet radiation (stress). Compared to the carrier, under stress conditions (e.g., ultraviolet exposure), the ingredient of this product in Renutriate PRO and Protectif Tinted significantly reduced the production of the key inflammatory marker interleukin-1β (IL-1β, Interleukin-1 beta). Figure 7A The presence of intracellular interleukin-1β messenger RNA (IL-1β mRNA) was assessed, and Figure 6BThe presence of intracellular interleukin-1β (IL-1β) protein was assessed. The amount of intracellular interleukin-1β messenger RNA (IL-1β mRNA) is correlated with the presence of intracellular interleukin-1β protein. Interleukin-1β triggers skin inflammation and irritation, often leading to collagen breakdown. This product's ingredients reduce interleukin-1β to alleviate inflammation, protect the skin from premature cellular aging, and create a soothing, healthy environment for skin regeneration. Inflammation is a hallmark of aging, and reducing interleukin-1β improves skin health and lifespan.

[0107] As further illustrated in Figure 8, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested in the presence or absence of UV radiation (stress). Compared to the carrier, the ingredient of this product in Renutriate PRO and Protectif Tinted significantly increased the production of DNA damage-induced transcription factor 4 messenger RNA (DDIT4 mRNA) under both stress-free and stress-induced conditions (e.g., UV exposure). Increased expression of DNA damage-induced transcription factor 4 (DDIT4) promotes cellular resistance to stress, particularly UV-induced damage. DDIT4 protein is involved in regulating the mTOR signaling pathway, which controls cellular metabolism and stress responses. DDIT4 is a key component in DNA repair mechanisms essential for repairing DNA damage or breaks caused by UV radiation. This product's ingredients increase the expression and protein production of DNA damage-induced transcription factor 4 (DDIT4), ensuring skin cells are better able to cope with environmental challenges, including UV radiation, and promoting repair and recovery. The ability to manage cellular stress is crucial for preventing damage accumulation. By increasing the production of DNA damage-induced transcription factor 4 (DDIT4), this product's ingredients significantly limit genomic instability and mitochondrial dysfunction, key markers of aging.

[0108] As further illustrated in Figure 9, epidermal cells were treated with a cream product or carrier containing this product's ingredients, and tested under conditions with and without UV radiation (stress). Compared to the carrier, the product's ingredients in Renutriate PRO and Protectif Tinted significantly increased the production of vascular endothelial growth factor (VEGF) mRNA under both stress-free and stress-related conditions (e.g., UV exposure). Stimulation of VEGF expression promotes new blood vessel formation, thereby improving skin radiance. VEGF is crucial for stimulating angiogenesis, ensuring adequate blood supply and oxygenation to the skin. This product's ingredients enhance skin healing and promote regeneration, particularly against damage caused by environmental factors such as UV radiation exposure, by stimulating VEGF expression. Upregulation of vascular endothelial growth factor (VEGF) combats mitochondrial dysfunction by improving oxygen delivery and supporting energy production in epidermal cells, promoting faster recovery and enhancing luster.

[0109] As further illustrated in Figure 10, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested with and without ultraviolet radiation (stress). Compared to the carrier, under stress conditions (e.g., ultraviolet exposure), the ingredient of this product in Renutriate PRO and Protectif Tinted significantly reduced the production of B-cell lymphoma-2 (BCL2) interacting protein 3 messenger RNA (BNIP3 mRNA). BCL2 interacting protein 3 expression is associated with cellular senescence and apoptosis. BCL2 interacting protein 3 regulates programmed cell death (apoptosis) and participates in cellular stress responses. This product's ingredients reduce BCL2 interacting protein 3 (BNIP3), delaying cell death and extending the lifespan of healthy epidermal cells, thereby ensuring overall improved skin function. Delayed apoptosis prevents premature cell death, a hallmark of cellular aging. Products containing this ingredient ensure skin cells continue to function optimally throughout the cell cycle.

[0110] As further illustrated in Figure 11, epidermal cells were treated with a cream product or carrier containing an ingredient of this product, and tested in the presence or absence of ultraviolet radiation (stress). Compared to the carrier, the ingredient of this product in Renutriate PRO and Protectif Tinted significantly increased the production of hypoxia-inducible factor-1α (HIF-1α) protein under both stress-free and stressful conditions (e.g., ultraviolet exposure). Increased expression of hypoxia-inducible factor-1α (HIF-1α) protein promotes angiogenesis and skin oxygenation. Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that responds to low oxygen levels by promoting the formation of new blood vessels (angiogenesis). This product's ingredients increase hypoxia-inducible factor-1α (HIF-1α) protein, improving oxygen and nutrient delivery to epidermal cells and promoting better healing and regeneration. Angiogenesis is crucial for maintaining skin vitality. Enhancement of hypoxia-inducible factor-1α (HIF-1α) can address mitochondrial dysfunction and loss of nutrient sensing, thereby improving overall skin health.

[0111] As further illustrated in Figure 12, epidermal cells were treated with a cream product or carrier containing this product ingredient, and tested with and without UV radiation (stress). Compared to the carrier, under stress conditions (e.g., UV exposure), the product ingredient in Renutriate PRO and Protectif Tinted significantly reduced the generation of reactive oxygen species (ROS) in epidermal cells. ROS cause oxidative damage to cells, a major factor leading to DNA damage, pigmentation, and skin aging. This product ingredient reduces ROS levels to prevent DNA damage and significantly reduce the chance of DNA mutations. Furthermore, reducing ROS levels protects skin structure. Oxidative stress accelerates aging by damaging DNA and cellular structure. The reduction of ROS by this product ingredient directly combats two key markers of aging: genomic instability and mitochondrial dysfunction.

[0112] This product's ingredients exhibit a comprehensive mechanism of action against twelve aging markers. Through multiple pathways, including but not limited to: collagen synthesis, extracellular matrix (ECM) remodeling, inflammation reduction, stress response modulation, and angiogenesis promotion. The combination of increased collagen production, enhanced ECM stability, reduced inflammatory markers, and improved stress resistance indicates that this product's ingredients can restore and maintain skin health.

[0113] Twelve signs of aging This product's ingredients help alleviate twelve signs of aging. By significantly increasing collagen synthesis and enhancing decorin messenger (DCN) expression, it reduces the loss of protein homeostasis. In doing so, it improves skin firmness, thereby reducing wrinkles and maintaining the balance of the extracellular matrix (ECM). It upregulates the production of laminin B1 (LAMB1), combating cellular aging and maintaining nuclear integrity. It reduces BCL2-interacting protein 3 (BNIP3) expression, delaying programmed cell death and thus extending cell lifespan, maintaining a youthful appearance. Reducing reactive oxygen species (ROS) levels under stress allows it to minimize DNA damage, further extending cell lifespan. Furthermore, it increases DNA damage-induced transcription factor 4 (DDIT4) expression, enhancing DNA repair mechanisms and protecting cellular DNA from mutations, thereby improving genome integrity. This product's ingredients also enhance laminin B1 (LAMB1), laminin B4 (LAMB4), and laminin A4 (LAMA4), which support nuclear structure and indirectly affect telomere stability. Maintaining telomere length contributes to cell lifespan. This product's ingredients have been shown to reduce reactive oxygen species (ROS) levels, thereby protecting mitochondria from oxidative stress. Furthermore, this product's ingredients regulate BCL2 interacting protein 3 (BNIP3) in epidermal cells, thereby influencing mitochondrial renewal and mitochondrial-related aging.

[0114] This product's components have also been shown to increase the transcription of DNA damage-induced transcription factor 4 (DDIT4), a component of metabolic pathway regulation under stress, thereby providing improved cellular metabolism. By enhancing extracellular matrix (ECM) components, this product's components can create a supportive environment in the epidermis for improved stem cell function and tissue repair. This product's components can reduce pro-inflammatory cytokines interleukin-6 (IL-6) and interleukin-1β (IL-1β), and increase hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF), thereby restoring cell signaling, improving tissue homeostasis, and reducing inflammation. This product's ingredients work by modulating BCL2 interacting protein 3 (BNIP3), thereby reducing autophagy, enhancing the cell cleansing process, and maintaining cell health.

[0115] The ingredients in this product have been shown to have the following effects: 1. Enhance skin structure and integrity; reduce chronic inflammation; 2. Improve cellular resistance and repair capabilities; and 3. Promote skin healing and radiance. Through collagen synthesis and extracellular matrix (ECM) remodeling (via prolyl 4-hydroxylase 1 (P4H1), laminin B1 (LAMB1), laminin B4 (LAMB4), laminin A4 (LAMA4), and decorin messenger (DCN)), the ingredients strengthen the structural components of the skin, preventing sagging, wrinkles, and loss of elasticity. The ingredients also reduce inflammatory cytokines (interleukin-6 (IL-6) and interleukin-1β (IL-1β),) thereby preventing chronic inflammation, a major cause of aging through tissue breakdown and degeneration. By enhancing stress response pathways (DNA damage-induced transcription factor 4 (DDIT4) and hypoxia-inducible factor-1 alpha (HIF-1α)), this product's ingredients promote angiogenesis, enabling skin cells to better resist environmental stressors such as UV damage and oxidative stress. Further promoting angiogenesis (vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1 alpha (HIF-1α)) allows the product's ingredients to improve blood flow and oxygenation in the epidermis, ensuring skin cells receive the nutrients needed for repair and regeneration, thereby improving skin texture and radiance. This product's multifaceted skincare approach targets core biological processes involved in aging. By responding to the skin's needs, this product's ingredients provide an advanced, adaptive solution for comprehensive anti-aging topical skincare compositions.

[0116] Tissue repair Using products containing this ingredient can significantly increase the expression of vascular endothelial growth factor (VEGF) under stress (UV exposure). VEGF plays a crucial role in angiogenesis and tissue repair, promoting the formation of new blood vessels and enhancing nutrient delivery to damaged tissues. This product's ingredients not only protect the skin but also actively enhance its regenerative capacity, a key factor in preventing and repairing UV-induced damage.

[0117] Activation of stress response pathways Using products containing this ingredient resulted in the upregulation of key molecular markers associated with cellular stress responses and cell survival. This ingredient increased the expression of DNA damage-induced transcription factor 4 (DDIT4), hypoxia-inducible factor-1α (HIF-1α), and prolyl 4-hydroxylase 1 (P4H1), factors crucial in mediating responses to hypoxia and oxidative stress. Hypoxia-inducible factor-1α (HIF-1α) is a master regulator of cellular responses to hypoxia, particularly important for maintaining cellular homeostasis and promoting survival under stress. Upregulation of the expression of DNA damage-induced transcription factor 4 (DDIT4), hypoxia-inducible factor-1α (HIF-1α), and prolyl 4-hydroxylase 1 (P4H1) enhanced the skin’s inherent repair mechanisms and reduced the long-term effects of UV-induced DNA damage.

[0118] Cellular structural integrity Using products containing this ingredient leads to the upregulation of laminin B1 (LAMB1), a key component of the nuclear lamina that is essential for maintaining nuclear structure and function. Analysis of epidermal cells has identified downregulation of laminin B1 (LAMB1) as a biomarker of cellular senescence. Cellular senescence has been shown to be associated with tissue aging and loss of function. This product's ingredient upregulates laminin B1 (LAMB1), not only preventing (or delaying) cellular senescence but also acting as an anti-aging agent, thereby promoting long-term skin health and resilience. This product's ingredient has been shown to enhance the skin's regenerative capacity.

[0119] Cancer Using products containing this ingredient can reduce or eliminate inflammation and oxidative stress, key drivers of aging and cancer. Significant reductions in pro-inflammatory cytokines interleukin-6 (IL-6) and IL-1β demonstrate the powerful anti-inflammatory effects of continued use of products containing this ingredient. Furthermore, application of this ingredient to the skin normalizes levels of reactive oxygen species (ROS) under UV exposure. The reduction in ROS and inflammatory markers highlights the potential of this ingredient in preventing chronic inflammation, a known contributing factor to premature skin aging and cancer.

[0120] extracellular matrix Using products containing this ingredient supports the maintenance and repair of the extracellular matrix, contributing to the structural integrity and function of the skin. UV exposure typically leads to a sharp decline in core proteoglycan (DCN, decorin messenger) levels, resulting in impaired collagen fiber production and skin structural damage. Using products containing this ingredient after UV exposure can increase DCN (decorin messenger) expression.

[0121] Erythematous acne and atopic dermatitis Sensitive skin conditions such as rosacea and atopic dermatitis can manifest as redness, inflammation, and varying degrees of discomfort due to epidermal damage. Rosacea is typically characterized by persistent redness, inflammatory papules, and flare-ups easily triggered by external factors such as the environment. Atopic dermatitis is characterized by red, itchy plaques on dry, scaly skin. Both conditions are generally challenging to treat, and inflammation further exacerbates this challenge.

[0122] Post-inflammatory hyperpigmentation The severity of post-inflammatory hyperpigmentation (“PIH”) varies depending on the underlying cause, such as psoriasis, lichen planus, or acne. Each condition presents unique challenges due to its inflammatory process and the resulting skin response. Regardless of the underlying cause, the ingredients in this product have been shown to effectively reduce post-inflammatory hyperpigmentation. Renutriate has been shown to significantly reduce hyperpigmentation lesions and deliver noticeable improvements in skin texture and uneven skin tone. After sixteen weeks of continuous treatment with Renutriate, post-inflammatory hyperpigmentation was reduced to a minimum, and in some cases even disappeared completely.

[0123] Patient Example 1 like Figures 13A to 13E Further description shows Patient 1's head. Patient 1 used Renutriate PRO, a professional repair and nourishing product containing ingredients from this product, twice daily for eight months. See details. Figure 13A The image shows the patient's skin condition on Day 0. Before using Renutriate PRO, Patient 1's skin had the following conditions: actinic keratosis, seborrheic keratosis, xerosis, and photodamage. These conditions could potentially lead to squamous cell carcinoma, basal cell carcinoma, pruritus, and seborrheic dermatitis. Over time, with continued use of Renutriate PRO, Patient 1's skin showed improvement. Actinic keratosis, seborrheic keratosis, xerosis, and photodamage almost completely subsided, and the skin's radiance increased, texture improved, and became smoother and more even-toned. See details for further information. Figure 13B Patient 1's skin showed significant improvement after 60 days of continuous use of a professional repair and nourishing product (Renutriate PRO) containing ingredients from this product. Actinic keratosis, seborrheic keratosis, xerosis, and photodamage on the skin were all reduced. See details... Figure 13C After 120 days of continuous use of Renutriate PRO, a professional repair and nourishing product containing ingredients from this product, Patient 1's skin showed further healing, with further reduction in actinic keratosis, seborrheic keratosis, xerosis, and photodamage. See details... Figure 13D Patient 1's skin showed further healing and regeneration. The skin appeared improved, healthier, younger-looking, and with a significantly enhanced radiance. (See details...) Figure 13EAfter 240 days of continuous use of Renutriate PRO, a professional repair and nourishing product containing this ingredient, Patient 1's skin exhibited a regenerated, healthy appearance. Actinic keratosis, seborrheic keratosis, xerosis, and photodamage almost completely subsided. Throughout Patient 1's use of Renutriate PRO, their skin showed increased collagen production and extracellular matrix (ECM) remodeling, resulting in improved skin texture and radiance. Furthermore, enhanced angiogenesis improved microcirculation, further enhancing skin radiance.

[0124] Patient Example 2 like Figures 14A to 14C Further description shows Patient 2's head. Patient 2 used Renutriate PRO, a professional repair and nourishing product containing ingredients from this product, twice daily for four months. See details. Figure 14A The image shows the patient's skin on day 0. Before using Renutriate PRO, Patient 2's skin exhibited the following conditions: actinic keratosis, seborrheic keratosis, photodamage, and wrinkles. These conditions could potentially lead to squamous cell carcinoma, basal cell carcinoma, itching, photodamage, and loss of elasticity and firmness. Over time, with continued use of Renutriate PRO, Patient 2's skin showed improvements in radiance and texture, becoming smoother and more even-toned. See details... Figure 14B After 60 days of continuous use of the professional repair and nourishing product (Renutriate PRO), Patient 2's skin appeared healthier, with reduced pitting and redness. See details... Figure 14C Patient 2's skin showed improved homeostasis after 120 days of continuous use of the Renutriate PRO professional repair and nourishing product, resulting in a healthier and more youthful appearance. Patient 2's skin exhibited a significant reduction in wrinkle depth and fine lines. The Renutriate PRO professional repair and nourishing product enhances collagen synthesis by increasing the expression of prolyl 4-hydroxylase subunit α1 (P4H1) and core proteoglycan (DCN, decorin messenger), contributing to firmer and smoother skin. Its effects are shown in [link to product description]. Figure 14C Skin becomes smoother and wrinkles are reduced.

[0125] Patient Example 3 like Figures 15A to 15CFurther description shows Patient 3's head. Patient 3 used Renutriate PRO, a professional repair and nourishing product containing ingredients from this product, twice daily for four months. See details. Figure 15A The image shows the patient's skin on day 0. Before using Renutriate PRO, patient 3's skin exhibited the following conditions: actinic keratosis, seborrheic keratosis, photodamage, and wrinkles. These conditions could potentially lead to squamous cell carcinoma, basal cell carcinoma, itching, photodamage, and loss of elasticity and firmness. Over time, with continued use of Renutriate PRO, patient 3's skin showed reduced signs of inflammation and a significant reduction in redness. See details... Figure 15B After 60 days of continuous use of the professional repair and nourishing product (Renutriate PRO), Patient 3 showed a reduction in signs of inflammation, actinic keratosis, and photodamage. Patient 3 also experienced a significant increase in skin firmness and elasticity. (See details...) Figure 15C The image shows the skin of Patient 3 after 120 days of continuous use of Renutriate PRO. Patient 3's skin showed a significant reduction in inflammation and redness. With continued use of Renutriate PRO, the patient's skin was noticeably firmer and less sagging. The decrease in interleukin-6 (IL-6) and interleukin-1β (IL-1β) confirms the anti-inflammatory properties of Renutriate PRO, making the skin more soothed and less prone to irritation.

[0126] Patient Example 4 like Figure 16A and 16B Further description shows the head of Patient 4. Patient 4 used Renutriate PRO, a professional repair and nourishing product containing ingredients from this product, twice daily for two months. See details. Figure 16A The image shows the patient's skin on day 00. Before using Renutriate PRO, Patient 4's skin exhibited the following conditions: actinic keratosis, photodamage, and severe wrinkles. These conditions could potentially lead to squamous cell carcinoma, basal cell carcinoma, itching, photodamage, loss of elasticity and firmness, and scaling and dryness. Over time, with continued use of Renutriate PRO, Patient 4's skin showed a reduction in redness and wrinkles. See details... Figure 16BPatient 4's skin showed progressive improvement in elasticity and firmness. With repeated use of the professional repair and nourishing product (Renutriate PRO), the patient's skin exhibited enhanced extracellular matrix (ECM) remodeling and restored skin firmness through the regulation of laminin B1 (LAMB1) and laminin B4 (LAMB4). The patient's skin also showed overall improvement in skin health and vitality, with a significant reduction in wrinkle depth.

[0127] Patient Example 5 like Figure 17A and 17B Further description shows Patient 5's legs. Patient 5 used Renutriate PRO, a professional repair and nourishing product containing ingredients from this product, twice daily for two months. See details. Figure 17A The image shows the patient's leg on day 0. Before using Renutriate PRO, patient 5's skin exhibited the following conditions: actinic keratosis, seborrheic keratosis, papular keratosis, retention keratosis, hypertrophic actinic keratosis, Bowen's disease, squamous cell carcinoma, edema, and stasis dermatitis. These conditions could potentially lead to: squamous cell carcinoma, basal cell carcinoma, pruritus, photodamage, loss of elasticity and firmness, peeling and dryness, venous stasis ulcers, impaired blood flow, and ischemia. Over time, with continued use of Renutriate PRO, signs of actinic keratosis, seborrheic keratosis, papular keratosis, retention keratosis, hypertrophic actinic keratosis, Bowen's disease, squamous cell carcinoma, edema, and stasis dermatitis decreased in patient 5. See details. Figure 17B Patient 5's skin showed a reduction in pigmentation and dark spots, resulting in a more even skin tone. The number and severity of actinic keratosis, hypertrophic actinic keratosis, seborrheic keratosis, and papular keratosis were significantly reduced. The patient's skin also showed significant reduction in inflammation, dryness / scaly skin, and edema. With continued use of Renutriate PRO, the patient's skin showed decreased levels of reactive oxygen species (ROS) and enhanced DNA repair mechanisms, manifested as a significant reduction in pigmentation. Furthermore, Renutriate PRO enhanced DNA damage repair, significantly reducing the burden of actinic keratosis. In summary, with continued use of Renutriate PRO, all aspects of skin health improved.

[0128] Patient Example 6 like Figure 18A and 18BFurther description shows the back of Patient 6. Patient 6 suffered moderate to severe sunburn due to excessive UV exposure. Following the sunburn, Patient 6 also experienced sunburn on his left shoulder (specifically as shown in the image). Figure 18A (As shown) Use Renutriate PRO, a professional repair and nourishing product containing ingredients of this product, twice daily for fourteen days. After sunburn, Patient 6 had sunburn on his right shoulder (specifically as shown). Figure 18B (As shown) Use a leading brand moisturizer twice daily for fourteen days. The left shoulder showed significantly better recovery from sunburn compared to the right shoulder. The left shoulder, treated with Renutriate PRO, recovered rapidly, with reduced redness, pigmentation, and peeling. The recovery observed on the left shoulder is the result of a synergistic effect between antioxidant protection (reduction of reactive oxygen species (ROS)), anti-inflammatory effects (reduction of interleukin-6 (IL-6) and interleukin-1β (IL-1β)), and angiogenesis (vascular endothelial growth factor (VEGF) F and hypoxia-inducible factor-1α (HIF-1α)). These effects accelerate skin healing, promote rapid recovery, and provide protection against UV damage.

[0129] Patient Example 7 like Figures 19A to 19C Further description shows the face of Patient 7. Patient 7 has a seven-year history of moderate rosacea, primarily affecting the cheeks and chin area. Patient 7 experiences intermittent symptoms of erythema, papules, and inflammatory bumps. These intermittent symptoms are further aggravated by sun exposure and stress. Patient 7 used a Renutriate product containing ingredients from this product twice daily for sixteen weeks. See details. Figure 19A The image shows the patient's face on day 0. Before using the Renutriate product, patient 7's skin exhibited the following characteristics: redness, papules, and inflammatory bumps. Over time, with continued use of the Renutriate product, patient 7's skin showed a reduction in signs of rosacea. See details... Figure 19B After eight weeks of continuous use of the repairing and nourishing product (Renutriate), Patient 7's skin showed a reduction in redness and dark spots, and a more even skin tone. The number and severity of rosacea significantly decreased. The patient's skin also showed a significant reduction in papules and inflammatory bumps. (See details...) Figure 19CThe image shows the patient's face at sixteen weeks. With continued use of Renutriate, the patient's skin showed a reduction in rosacea symptoms, with a significant decrease in inflammation, papules, and redness. The patient's skin exhibited a noticeably soothed and smoother appearance, with only slight visible redness remaining. The ingredients in Renutriate alleviate rosacea symptoms by modulating and downregulating inflammatory cytokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and CCL2. These cytokines are crucial for rosacea development and have been shown to be reduced by the ingredients in this product.

[0130] Patient Example 8 like Figures 20A to 20C Further description shows Patient 8's hands. Patient 8 has a two-year history of moderate atopic dermatitis, primarily affecting the hands. Patient 8 experiences intermittent flare-ups during the winter, leading to cracked and flaky skin. Intermittent symptoms manifest as itching, dryness, and inflammation. Patient 8 used a Renutriate product containing ingredients from this product twice daily for two weeks. See details. Figure 20A The image shows the patient's face on day 0. Before using Renutriate, patient 7's skin exhibited the following conditions: itching, cracking, and dryness around the knuckles, interphalangeal joints, and finger webs. Over time, with continued use of Renutriate, patient 8's skin showed signs of improvement in atopic dermatitis. See details... Figure 20B After one week of continuous use of the repairing and nourishing product (Renutriate), patient 8 experienced a reduction in the severity of redness and peeling. Itching significantly decreased, and visible skin cracks began to heal. (See details...) Figure 20C The image shows the patient's hands after two weeks. With continued use of Renutriate, the patient's skin showed further reduction in dryness and inflammation. The patient's skin presented a significantly more soothed appearance, with only very slight residual redness and flaking. The active ingredient in Renutriate can alleviate symptoms of atopic dermatitis by modulating inflammatory pathways, possibly by downregulating the CCL-2-CCR2 pathway.

[0131] Patient Example 9 like Figures 21A to 21D Further description shows Patient 9's ankle. Patient 9 has a ten-year history of psoriasis, primarily affecting the lower extremities. Patient 9 used a Renutriate product containing ingredients from this product twice daily for sixteen weeks. See details... Figure 21AThe image shows the patient's ankle on day 0. Patient 9's skin before using the Renutriate product showed well-defined, hyperpigmented patches, ranging in color from dark brown to grayish-brown. See details... Figure 21B After eight weeks of continuous use of the repairing and nourishing product (Renutriate), patient 9's skin showed a significant reduction in hyperpigmentation, and the surrounding skin showed reduced scaling and a smoother appearance. (See details...) Figure 21C The image shows the patient's skin at week twelve. The patient's skin showed further reduction in hyperpigmentation, a more even skin tone, appeared more elastic, and felt smoother overall. See details for further information. Figure 21D The image shows the patient's skin after sixteen weeks of continuous use of the Renutriate product. The patient's skin showed very mild hyperpigmentation, and the hyperpigmented patches had almost completely faded. The patient's skin appeared healthy and balanced.

[0132] Patient Example 10 like Figures 22A to 22C Further description shows Patient 10's hands. Patient 10 has a five-year history of lichen planus affecting their hands, neck, and chest. Patient 10 used a Renutriate product containing ingredients from this product twice daily for sixteen weeks. See details. Figure 20A The image shows the patient's hand on day 0. Before using the Renutriate product, patient 10's skin exhibited well-defined, grayish-brown patches of hyperpigmentation. With continued use of the Renutriate product over time, patient 10's skin showed a significant reduction in hyperpigmentation. See details... Figure 22B The image shows the patient's hands after eight weeks of continuous use of the Renutriate repair product. The patient's hands showed a significant reduction in hyperpigmentation, and the surrounding skin exhibited reduced scaling and a smoother appearance. See details for further information. Figure 22C The image shows the patient's skin after sixteen weeks of continuous use of the Renutriate product. The patient's skin showed very little pigmentation, and the hyperpigmented patches had almost completely faded, leaving the skin looking healthy and balanced.

[0133] Patient Example 11 like Figures 23A to 23C Further description shows the face of Patient 11. Patient 11 has a one-year history of facial acne vulgaris. Patient 11 used Renutriate PRO, a professional repair and nourishing product containing ingredients of this product, twice daily for sixteen weeks. See details. Figure 23AThe image shows the patient's face on day 0. Before using the Renutriate product, patient 11's skin exhibited acne and well-defined hyperpigmentation surrounding previous acne lesions. Over time, with continued use of the Renutriate product, patient 11's skin showed a reduction in signs of acne and hyperpigmentation. See details... Figure 23B After eight weeks of continuous use of the professional repair and nourishing product (Renutriate PRO), patient 11 showed a reduction in the severity of acne and hyperpigmentation. (See details...) Figure 23C The image shows the patient's face at sixteen weeks. With continued use of Renutriate PRO, the patient's skin showed further reduction in acne and hyperpigmentation, with very few acne lesions remaining and the skin almost completely clear. The skin looked healthier and more vibrant. Furthermore, the patient's hyperpigmentation was mild, and hyperpigmented patches almost completely disappeared. The ingredients in Renutriate help reduce symptoms of acne and hyperpigmentation.

[0134] Reversing the aging process Renutriate PRO and Protectif Tinted products, when applied to the skin, have shown not only to slow the rate of premature cell aging but also to reverse the aging process. In vitro and in vivo analyses of epidermal cells coated with Renutriate PRO or Protectif Tinted revealed significant effects on several key cellular components.

[0135] As shown in Figures 6, 7, and 11, epidermal cells coated with Renutriate PRO and Protectif Tinted exhibited significant effects on key cellular components. Figures 6, 7, and 11 illustrate the regulation of key cellular component expression under environmental stressors. Upon exposure to environmental stressors, epidermal cells coated with Renutriate PRO and Protectif Tinted showed: a two-thirds reduction in interleukin-6 (IL6) expression (Figure 6), a 50% reduction in interleukin-1β (IL-1β) expression (Figure 7), and a five-fold increase in hypoxia-inducible factor-1 alpha (HIF-1α) expression (Figure 11). As key cellular components expressed by environmental stressors, interleukin-6 (IL6), interleukin-1β (IL-1β), and hypoxia-inducible factor-1α (HIF-1α) also play crucial roles in premature cellular aging. Environmental stressors induce the regulation of key intracellular components, which in turn contributes to premature cellular aging.

[0136] Oxidative stress is another environmental stressor that plays a significant role in premature aging. The effects of oxidative stress and the resulting formation of reactive oxygen species (ROS) have a destructive impact on the skin and have been shown to be a key factor in cellular aging. ROS have also been shown to significantly increase the rate of premature skin aging. As further illustrated in Figure 12, in the presence of environmental stressors, epidermal cells incubated with Renutriate PRO showed a 3-fold reduction in ROS production. This reduction was very significant, restoring ROS production to its initial level (the level without environmental stressors).

[0137] Renutriate PRO has been tested on numerous patients and has shown significant modulation of the expression of key cellular components in response to environmental stressors. This response to environmental stressors not only enables epidermal cells to effectively combat the effects of environmental stressors but also significantly slows down the rate of premature aging. Therefore, research suggests that using Renutriate PRO can reduce the overall age of the skin.

[0138] Patient Example 12 Patient 12, a 47-year-old male, had been proactively using Renutriate PRO and Protectif Fair Tint once daily on his left forearm for nine months. He had also been proactively using a leading brand skincare product (moisturizer and sunscreen) once daily on his right forearm for nine months. Patient 12 had an active lifestyle, and both forearms were frequently exposed to environmental stressors. The relative age of Patient 12's left and right forearms was then measured, directly related to the effects of environmental stressors on premature aging of epidermal cells. Skin age was measured using a validated Total Antioxidant Capacity (PAOT) skin scan device (“PAOT”). PAOT is a clinically validated tool that measures oxidative stress and reactive oxygen species (ROS) levels in epidermal cells in real time, providing a true skin age. In addition to assessing oxidative stress in the skin, PAOT can also test the following indicators: a. Vitamin C ascorbic acid; b. Skin hydration; c. pH balance; d. Glutathione (GSH); and e. Vitamin E tocopherol. See details... Figure 24A Patient 12's left forearm (treated with Renutriate PRO and Protectif Fair Tint) had a skin age of 23.7. See details for further information. Figure 24BPatient 12's right forearm (treated with leading brand skincare products) had a skin age of 53.8. The use of Renutriate PRO and Protectif Fair Tint not only significantly reduced premature aging but also lowered the skin age to below Patient 12's actual age (47 years). Patient 12 demonstrated that Renutriate PRO and Protectif Fair Tint can regulate epidermal cells, not only slowing down the aging process but even reversing it. PAOT also showed differences between the left and right forearms in vitamin C, vitamin E, hydration, pH balance, and total reduced glutathione levels. The effect observed in the left forearm of patient 12 was attributed not only to the reduction in oxidative stress, but also to the increased expression of DNA damage-induced transcription factor 4 (DDIT4) (Figure 8), a DNA repair enzyme that helps protect epidermal cells from UV-induced DNA damage and the resulting erroneous production of cellular proteins and enzymes.

[0139] The ultrasound images presented in Figure 24 compare and assess skin collagen density and extracellular matrix (ECM) integrity under two different skincare regimens. From a dermatological and scientific perspective, these ultrasound images reveal significant differences in the structure, organization, and density of the dermis. The right image shows relatively low collagen fiber density with visible breaks and reduced uniformity, indicating poor extracellular matrix support. This suggests that while leading brand skincare products (i.e., moisturizers and sunscreens) provide a baseline level of protection, they do not meaningfully stimulate dermal regeneration or enhance collagen integrity. Conversely, the left image—representing skin treated with RenutriatePRO and Protectif Fair Tint—shows a significant increase in dermal density and a more organized and cohesive collagen network. This is evidenced by enhanced ultrasound echogenicity (brightness and density) in the ultrasound scans, which is associated with higher collagen deposition, improved extracellular matrix (ECM) remodeling, and better skin biomechanical properties. This denser, more uniform structure indicates that Renutriate PRO and Protectif Fair Tint actively promote fibroblast proliferation, collagen synthesis, and extracellular matrix (ECM) stabilization, resulting in visible and functional improvements in skin elasticity, hydration, and overall anti-aging effects. Unlike leading brand skincare products that primarily act as a passive barrier, the ingredients in Renutriate PRO and Protectif Fair Tint work actively at the cellular level, stimulating collagen production, enhancing skin structural integrity, and accelerating tissue regeneration (the skin's own repair mechanism). Significant differences in ultrasound imaging highlight the efficacy of Renutriate PRO and Protectif Fair Tint in reversing age-related dermal thinning, enhancing skin firmness, and restoring youthful elasticity.

[0140] Patient Example 13 For details, please refer to the following: Figure 25APatient 13, a 47-year-old male, had Renutriate PRO applied to location 1 and a leading brand skincare product applied to location 2 on his biceps (the "test area") for 20 minutes. After the 20-minute skincare incubation period, the test areas were cleansed and irradiated with a UVB lamp (Mx1000 UVB lamp) for 75 seconds to apply UVB stress. Following the UVB stress, the skin age at locations 1 and 2 was assessed using PAOT testing. Reference Figure 25B and Figure 25C PAOT analysis showed that epidermal cells at location 1 were significantly younger than those at location 2. The results indicate that using Renutriate PRO for just twenty minutes significantly affected the regulation of epidermal cell component expression, thereby mitigating the effects of UVB stress exposure and significantly reducing oxidative stress and the production of reactive oxygen species (ROS).

[0141] Patient Example 14 Patient 14, a 68-year-old woman, had been regularly using a leading brand of skincare products on her face for three years. Her skin age was tested before and after using Renutriate PRO. Patient 14's skin was tested using PAOT before using Renutriate PRO to provide a baseline of her skin age. After applying Renutriate PRO to her face, Patient 14 waited ninety (90) minutes before testing her skin age again using PAOT. Patient 14's skin age was significantly reduced after using Renutriate PRO. The results indicate that Renutriate PRO effectively reduces skin age regardless of the quality and duration of the skincare regimen.

[0142] In one embodiment, the novel topical skin composition comprises at least one selected from hydroxytyrosol, green tea extract, and green coffee extract. The concentration of hydroxytyrosol in the topical skin composition can be between 0.1% w / w (weight / weight) and 0.75% w / w. The concentration of green tea extract in the topical skin composition can be between 0.025% w / w and 0.1% w / w. The concentration of green coffee extract in the topical skin composition can be between 0.025% w / w and 0.25% w / w. In a preferred embodiment, the green coffee extract can be between 0.025% w / w and less than 0.2% w / w. In one embodiment, the green coffee extract is a green coffee bean extract. The green coffee bean extract can be an Arabica coffee bean extract. The green tea extract can be a green tea leaf extract, and in one embodiment of this disclosure, the green tea leaf extract is a Camellia sinensis leaf extract.

Claims

1. A method for activating skin cells, the method comprising: A topical skin composition is provided, the composition comprising: Hydroxytyrosol at concentrations ranging from 0.1% w / w to 0.75% w / w. Green tea extract at concentrations ranging from 0.025% w / w to 0.1% w / w; and Green coffee bean extract at concentrations ranging from 0.025% w / w to less than 0.2% w / w; and The topical skin composition is applied topically to the skin.

2. The method for activating skin cells according to claim 1, wherein the method for activating skin cells is used to prevent skin cell aging, and wherein the topical skin composition produces at least one of the following effects: Increase DDIT4 expression in cells to promote DNA repair in skin epidermal cells; It reduces reactive oxygen species levels and increases HIF-1α production to enhance oxidative stress responses in skin epidermal cells; Stimulating P4H1 cell expression to enhance collagen biosynthesis in skin epidermal cells; Increase the cellular expression of at least one of LAMB1, LAMB4, LAMA4 and DCN to enhance extracellular matrix remodeling of skin epidermal cells; Reduce cellular expression of at least one of IL6 and IL1β to reduce inflammatory response to external stressors; Increase VEGF expression in cells to increase angiogenesis; and Reduce BNIP3 expression in cells to reduce apoptosis in skin epidermal cells.

3. The method for activating skin cells according to claim 1, wherein the method for activating skin cells is used to prevent cell aging, and wherein the topical skin composition produces at least one of the following effects: Increases LAMB1 expression in skin epidermal cells; and, Reduce BNIP3 expression in cells.

4. The method for activating skin cells according to claim 1, wherein the method for activating skin cells is used to reverse the aging process within the skin, wherein the topical skin composition produces at least one of the following effects: Increase DDIT4 expression in cells to promote DNA repair in skin epidermal cells; Lowering reactive oxygen species levels and increasing HIF-1α production to enhance oxidative stress response in skin epidermal cells; and Reduce cellular expression of at least one of IL6 and IL1β to alleviate the inflammatory response to external stressors.

5. The method for activating skin cells according to claim 1, wherein the topical skin composition is applied twice daily for 14 days to 8 months.

6. The method for activating skin cells according to claim 1, wherein the topical skin composition is applied once daily for 9 months.

7. The method for activating skin cells according to claim 1, wherein the topical skin composition is applied once for 20 minutes.

8. A topical skin composition comprising: Hydroxytyrosol at concentrations ranging from 0.1% w / w to 0.75% w / w. Green tea extract at concentrations ranging from 0.025% w / w to 0.1% w / w; and Green coffee bean extract at concentrations of 0.025% w / w to less than 0.2% w / w. The topical skin composition is applied to the surface of epidermal cells to modulate the cellular gene expression profile, which is crucial for skin health, prevention of skin diseases, and reduction of the severity of affected skin cells.

9. The topical skin composition of claim 8, wherein the topical skin composition increases the total amount of key cellular components, thereby restoring the skin to its natural healthy state while enhancing the skin's natural repair and defense mechanisms.

10. The topical skin composition of claim 8, wherein the topical skin composition counteracts the negative effects of environmental stressors, inflammation, and autoimmune diseases.

11. The topical skin composition according to claim 8, wherein the modulation of the cell gene expression profile produces at least one of the following effects: Reduces IL-6 in epidermal cells; Increase DDIT4 in epidermal cells; Increase P4H1 in epidermal cells; Increase VEGF in epidermal cells; and Reduce BNIP3 in epidermal cells.

12. The topical skin composition according to claim 8, used to regulate cellular gene expression to treat a variety of epidermal diseases and reduce the severity of symptoms.

13. The topical skin composition according to claim 12, when applied to the skin surface twice daily, is effective in treating at least one of actinic keratosis, seborrheic keratosis, steatosis, retention keratosis, hypertrophic actinic keratosis, xerosis, and photodamage.

14. The topical skin composition of claim 12, when applied to the skin surface twice daily, has a therapeutic effect on rosacea.

15. The topical skin composition of claim 12, when applied to the skin surface twice daily, has the effect of treating and reducing the likelihood of at least one of the following diseases: squamous cell carcinoma and basal cell carcinoma.

16. The topical skin composition of claim 12, when applied to the skin surface twice daily, has therapeutic effects on at least one of the following conditions: seborrheic dermatitis, venous stasis dermatitis, Bowen's disease, edema, venous stasis ulcers, wrinkles, impaired blood flow and ischemia, and pruritus.

17. A method for treating one or more of aging markers, skin degeneration, collagen, and elastin in the form of a single-dose pharmaceutical composition, said method comprising: Provided a pharmaceutical composition comprising: Hydroxytyrosol at concentrations ranging from 0.1% w / w to 0.75% w / w. Green tea extract at concentrations ranging from 0.025% w / w to 0.1% w / w; and Green coffee bean extract at concentrations ranging from 0.025% w / w to less than 0.2% w / w; and The pharmaceutical composition is applied topically to the skin.

18. The treatment method according to claim 17, wherein the pharmaceutical composition produces at least one of the following effects: Increase DDIT4 expression in cells to promote DNA repair in skin epidermal cells; It reduces reactive oxygen species levels and increases HIF-1α production to enhance oxidative stress responses in skin epidermal cells; Stimulating P4H1 cell expression to enhance collagen biosynthesis in skin epidermal cells; Increase the cellular expression of at least one of LAMB1, LAMB4, LAMA4 and DCN to enhance extracellular matrix remodeling in skin epidermal cells; Reduce cellular expression of at least one of IL6 and IL1β to alleviate the inflammatory response to external stressors; Increase VEGF expression in cells to increase angiogenesis; and Reduce BNIP3 expression in cells to decrease apoptosis of skin epidermal cells.

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