A soothing composition for repairing and uses thereof

Abstract

This invention relates to the field of cosmetics, and more particularly to a repairing and soothing composition and its application. The invention provides a repairing and soothing composition comprising: panthenol, niacinamide, and grape flower cell extract; the mass ratio of panthenol, niacinamide, and grape flower cell extract is (0.01~5):(0.01~5):(0.00001~0.3). This invention uses panthenol, niacinamide, and grape flower cell extract in combination, with each component working synergistically to enhance the expression of barrier-related proteins and inhibit the expression of inflammatory factors. This achieves the effects of repairing the damaged skin barrier after cosmetic procedures and soothing post-operative inflammatory pigmentation, meeting the urgent repair and soothing needs of modern people after cosmetic procedures.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, and more particularly to a repairing and soothing composition and its application. Background Technology

[0002] Post-medical aesthetic care is a crucial part of skincare, directly impacting the presentation and maintenance of treatment results. However, while laser, radiofrequency, and microneedling treatments improve skin problems, they can also cause varying degrees of temporary damage to the skin barrier, leading to post-operative reactions such as redness, burning, dryness, and peeling. Furthermore, improper post-operative care, environmental irritants, and individual differences can all affect the skin's repair process, even causing complications such as pigmentation and infection. With the rapid development of medical aesthetic technology and consumers' increasing demands for post-operative results, how to scientifically alleviate post-operative discomfort and rebuild a healthy skin barrier has become a common concern for both consumers and practitioners in the field.

[0003] The current market for post-cosmetic surgery skincare products faces significant challenges: while procedures like IPL and microneedling can effectively improve skin texture, they often result in skin barrier damage and inflammatory reactions. Worse still, post-inflammatory hyperpigmentation frequently leads to post-procedure darkening. Traditional post-operative skincare solutions often suffer from slow results (e.g., ordinary moisturizers), high irritation (e.g., high-concentration anti-inflammatory ingredients), or incomplete repair, leading to prolonged recovery periods and potentially causing secondary sensitivity.

[0004] Therefore, it is necessary to provide a product that repairs the skin barrier and soothes discomfort to meet people's complex and urgent skincare needs after cosmetic procedures. Summary of the Invention

[0005] In view of this, the present invention provides a repairing and soothing composition and its application. The present invention uses panthenol, nicotinamide, and grape flower cell extract in combination. These components work synergistically and promote each other, exhibiting a synergistic effect in promoting the expression of barrier-related proteins and inhibiting the expression of inflammatory factors. This achieves the effects of repairing the damaged skin barrier after cosmetic procedures and soothing post-operative inflammatory pigmentation, meeting the urgent repair and soothing needs of modern people after cosmetic procedures.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a repairing and soothing composition comprising: panthenol, nicotinamide, and grape flower cell extract; The mass ratio of the panthenol, the nicotinamide, and the grape flower cell extract is (0.01~5):(0.01~5):(0.00001~0.3).

[0007] In some embodiments of the present invention, the mass ratio of panthenol, nicotinamide and grape flower cell extract in the above-mentioned repairing and soothing composition is (0.01~0.5):(0.01~5):(0.00001~0.3).

[0008] In some embodiments of the present invention, the mass ratio of panthenol, nicotinamide and grape flower cell extract in the above-mentioned repairing and soothing composition is (0.03~3):(0.03~3):(0.001~0.1).

[0009] In some embodiments of the present invention, the mass ratio of panthenol, nicotinamide and grape flower cell extract in the above-mentioned repairing and soothing composition is 0.3:3:0.1.

[0010] The present invention also provides the use of the above-described repairing and soothing composition in the preparation of anti-inflammatory, repairing and / or soothing products.

[0011] In some embodiments of the present invention, the above applications are used after medical aesthetic procedures, including but not limited to one or more of phototherapy procedures such as photofacial rejuvenation, non-ablative laser, and radiofrequency.

[0012] In some embodiments of the present invention, the above-described applications involve the product being used after cosmetic surgery.

[0013] The present invention also provides the use of the above-described repair and soothing composition in the preparation of products that upregulate the expression of the IVL gene.

[0014] The present invention also provides the use of the above-described repairing and soothing composition in the preparation of products that inhibit and / or reduce the expression of inflammatory factors.

[0015] In some embodiments of the present invention, the inflammatory factor described above includes IL-6.

[0016] The present invention also provides the use of the above-described repair and soothing composition in the preparation of products that upregulate the expression of FLG and / or LOR genes.

[0017] The present invention also provides a product comprising: the above-described repairing and soothing composition.

[0018] In some embodiments of the present invention, the above-mentioned products include: personal care products.

[0019] In some embodiments of the present invention, the above-mentioned products include: post-medical aesthetic surgery repair and soothing products.

[0020] In some embodiments of the present invention, the personal care product described above includes: a basic care product, which includes, but is not limited to: Cleansing milk and / or facial wash: Uses a mild surfactant system, optionally combined with amino acid surfactants or betaine surfactants, to gently remove facial dirt, excess oil and a small amount of makeup residue during the vulnerable period after cosmetic procedures, minimizing further damage to the skin's lipid barrier; Cleansing oils and / or cleansing waters and / or cleansing balms: specially designed to thoroughly remove makeup and physical sunscreens, especially suitable for removing waterproof cosmetics; preferably using low-irritant, low-allergenic oil-phase or micellar formulas to reduce post-treatment skin friction and irritation; Toner and / or lotion and / or softening lotion: Used after cleansing, it can further remove trace residues on the skin surface, while replenishing the skin with moisture and helping to restore the pH balance that has been disrupted by treatments such as phototherapy and chemical peels, creating a favorable environment for the penetration and absorption of subsequent repair ingredients. Serum: Contains high concentrations of active ingredients, such as peptides, stylosin, and vitamin C derivatives, which can provide deep nourishment, soothing and repair for common problems after cosmetic procedures, such as redness, dryness, fine lines, and dullness. Eye cream and / or eye serum: specially designed for the thin and sensitive skin around the eyes, it has the effects of reducing fine lines and wrinkles, relieving dark circles and puffiness around the eyes, and firming the eye contour. The texture is usually light, easy to spread and easy to absorb, avoiding postoperative pulling and burden. Daytime lotion and / or cream: Used during the day, it has moisturizing, barrier repair, and antioxidant functions. It can form a breathable protective film on the skin surface, reduce the stimulation of external pollutants, dust and microorganisms on the postoperative wound, and help lock in moisture. Nighttime lotion and / or cream: Used at night, focusing on nighttime repair and cell renewal. It can be combined with barrier repair ingredients such as ceramides, squalane, and phytosterols to promote epidermal cell regeneration and barrier function reconstruction, and improve postoperative dryness, peeling and discomfort. Sunscreen and / or physical sunscreens: Used for postoperative recovery and daily UV protection. Highly stable, safe, and low-allergenic physical sunscreens (such as nano- or non-nano-sized titanium dioxide and zinc oxide) are preferred to prevent post-inflammatory hyperpigmentation and photoaging and reduce the risk of pigmentation. Facial masks and / or sheet masks and / or cream masks and / or freeze-dried powder masks: provide concentrated, short-term intensive care for the skin, such as hydration, soothing and calming, pore cleansing or brightening, especially suitable for short-term calming and hydration after surgery. Formulas that do not contain alcohol, fragrance and strong preservatives can be selected. Ampoules and / or single-use serums: These use sterile, small-dose, single-use packaging and are rich in high-purity, highly active repairing ingredients, such as asiaticoside, ectoine, hyaluronic acid and its cross-linked polymers, for intensive repair during the acute or sensitive period after surgery, reducing contamination and secondary irritation. Repair spray and / or soothing spray: Based on mineral water, hot spring water, sea spring water or water containing soothing ingredients (such as panthenol, allantoin, dipotassium glycyrrhizate), it can be sprayed multiple times after the operation to provide immediate hydration, cooling and soothing of stinging and burning, while reducing the risk of infection from touching the face with hands. Serums and / or single-ingredient essences: These are based on a single high-purity active ingredient (such as hyaluronic acid serum, niacinamide serum, arbutin serum, etc.) and can be used in combination according to the post-operative skin condition to specifically address dryness, dehydration, dullness, or pigmentation. Pre-serum and / or pre-serum: Used before serum, these products enhance the transdermal absorption efficiency of subsequent repair ingredients through small molecule penetration enhancers or penetration-enhancing technologies, thereby improving the overall repair effect of the skincare routine. Massage cream and / or massage gel: After the skin barrier has basically recovered, use gentle techniques to massage the face to promote local microcirculation and lymphatic drainage, which helps to reduce postoperative swelling and soften facial contours. Low-irritant and easy-to-wash bases are usually selected. Sleeping masks and / or overnight gel masks: Used as the last step in nighttime skincare, they form a breathable film that continuously and slowly releases moisturizing and repairing ingredients, extending the nighttime repair time. Suitable for use during the recovery period after surgery when dryness and tightness are obvious. Lip scrub and / or lip mask: To address the dryness and peeling of lips that may occur after the procedure, it improves roughness and chapping of lips through gentle exfoliation or occlusive repair, and enhances the softness and comfort of lips. Neck care cream and / or neck mask: Specifically designed for the thin and easily sagging skin of the neck, providing moisturizing, firming and wrinkle-reducing care to prevent and improve dry lines and sagging of the neck caused by improper care after medical aesthetic treatments (such as radiofrequency and ultrasound procedures); Hand care cream and / or hand mask: Used for moisturizing and barrier repair of postoperative hand skin, especially in situations where disinfection and hand washing are frequently required, to reduce postoperative hand dryness, cracking and sensitivity; Body lotion and / or body cream and / or body serum: for daily moisturizing and repair of the skin below the neck, especially suitable for large-area skin care after laser hair removal and body tightening treatments, relieving dryness and itching; Feminine hygiene wash and / or feminine moisturizing gel: Gentle cleansing and moisturizing for sensitive areas such as the perineum and groin. It uses a weakly acidic, soap-free, and low-irritant formula to reduce postoperative discomfort and irritation risk. Repairing essential oils and / or skincare oils: Based on plant oils (such as jojoba oil, rosehip oil, sunflower seed oil) or synthetic esters, they are compounded with fat-soluble repairing ingredients (such as vitamin E and phytosterols) to provide oil-sealing repair when the skin barrier is severely damaged, reducing transepidermal moisture loss; Lyophilized powder and / or solvent powder formulations: Highly active and unstable repair ingredients (such as growth factors, peptides, probiotic fermentation products, etc.) are preserved in lyophilized form and mixed with a special solvent before use to ensure activity and safety for use in the acute postoperative period. Medical dressings and / or cold compresses and / or collagen patches: These are products that fall under the category of medical devices or are between cosmetics and medical devices. They are produced using sterile or low-bacterial processes and their main components are collagen, hyaluronic acid, carbomer, etc. They are used for postoperative wound covering, cold compress cooling, moisturizing and isolation, and promoting healing. Sunscreen and / or makeup primer (post-operative type): Use after your doctor has given permission to resume makeup application. It is formulated with mineral-based, hypoallergenic, and non-comedogenic ingredients to isolate makeup from external environmental irritants on delicate skin, while also providing some color correction and evening out skin tone. Setting spray (gentle): Used for setting makeup during the post-operative recovery makeup stage. It is formulated with alcohol-free and low-irritant ingredients to reduce friction and irritation to the skin caused by frequent makeup touch-ups. Scalp care essence and / or hairline care solution: Targeting the post-operative care needs of hair transplantation, scalp microneedling, etc., it provides scalp soothing, moisturizing and hair follicle nutrition support, reducing post-operative scalp dryness, itching and discomfort; Scar removal gels and / or silicone gels: used for scar management after invasive cosmetic procedures (such as surgical excision, fractional CO2 laser, etc.), preventing and reducing hypertrophic scars and pigmentation changes by sealing hydration and regulating collagen deposition; Soothing repair sticks and / or topical gels: for precise care of postoperative local redness, swelling, papules or small wounds, easy to carry and reapply anytime, minimizing impact on surrounding normal skin; Repairing cotton pads and / or wipes (non-woven fabric products): Made of pure cotton or non-woven fabric, soaked in a mild, low-irritant cleansing or soothing solution, used for gentle wiping and temporary care when it is inconvenient to clean with water after surgery.

[0021] The beneficial effects of this invention include: (1) The composition of the present invention is precisely targeted at the post-medical aesthetic treatment scenario: After medical aesthetic treatment, the skin is in a state of high sensitivity or high demand. Ordinary skin care products or ingredients may cause irritation, and existing drug preparations often over-treat. The composition of the present invention is designed specifically for this scenario. The ingredients are simple and the target is clear. It can meet the needs of barrier repair and anti-inflammatory soothing without increasing the burden on the skin.

[0022] (2) The components of the composition described in this invention have a clear synergistic effect mechanism: for the first time, nicotinamide, panthenol, and grape flower cell extract are combined. Panthenol can be converted into pantothenic acid, promoting fibroblast proliferation and epithelialization; nicotinamide promotes ceramide synthesis and enhances skin barrier function; grape flower extract enhances epidermal moisture retention and antimicrobial peptide expression, forming a multi-pathway synergistic system of physical barrier repair, moisturizing and healing, and chemical barrier enhancement. Therefore, the above components can achieve barrier repair in multiple ways, exert synergistic effects, and at the same time, relieve discomfort by inhibiting inflammatory factor pathways, breaking through the single-effect limitations of traditional products and meeting the complex and urgent repair needs of people after medical aesthetic procedures.

[0023] (3) The present invention provides a preferred ratio of panthenol, nicotinamide and active ingredients of grape flower cell extract: 0.03~0.3:0.03~3:0.001~0.1. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0025] Figure 1 This indicates the relative expression level of the IVL gene; Figure 2 This indicates the relative expression levels of LOR genes; Figure 3 This indicates the relative expression level of the FLG gene; Figure 4 The relative expression level of the IL-6 gene is shown. Detailed Implementation

[0026] This invention discloses a repairing and soothing composition and its application.

[0027] It should be understood that the expression “one or more of…” individually includes each of the objects described after the expression, as well as various different combinations of two or more of the described objects, unless otherwise understood from the context and usage. The expression “and / or” combined with three or more described objects should be understood to have the same meaning, unless otherwise understood from the context.

[0028] The terms “including,” “having,” or “containing,” including the use of their grammatical synonyms, should generally be understood as open-ended and non-restrictive, for example, not excluding other unstated elements or steps, unless otherwise specifically stated or understood from the context.

[0029] It should be understood that the order of the steps or the order in which certain actions are performed is not important as long as the invention remains operational. Furthermore, two or more steps or actions can be performed simultaneously.

[0030] The use of any and all instances or exemplary language such as “e.g.” or “including” in this document is merely intended to better illustrate the invention and is not intended to limit the scope of the invention unless the claims are made. No language in this specification should be construed as indicating that any unclaimed element is essential to the practice of the invention.

[0031] Furthermore, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Therefore, unless explicitly stated otherwise, it should be understood that all ranges, quantities, values, and percentages used in this disclosure are modified with the word "approximately". Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range.

[0032] In a first aspect, the present invention provides a composition for post-medical aesthetic repair and soothing, the composition comprising: panthenol, nicotinamide and grape flower cell extract.

[0033] In this invention, the effects of each component are as follows: Panthenol, also known as panthenol or pro-vitamin B5, is a precursor to coenzyme A. It can be converted into pantothenic acid to participate in energy metabolism, providing energy for the repair of damaged skin. After penetrating the stratum corneum, panthenol stimulates hyaluronic acid synthesis, significantly improving skin hydration and relieving dryness. In the early stages of inflammation, panthenol can rapidly inhibit inflammatory signaling pathways, reduce the release of inflammatory factors such as prostaglandin E2, alleviate skin redness and stinging symptoms, accelerate the synthesis of lipids such as ceramides and cholesterol in the skin barrier, strengthen the skin's "brick wall structure," and enhance the skin's ability to resist external stimuli.

[0034] Niacinamide, also known as nicotinamide, is an amide compound of nicotinic acid (VB3). It directly strengthens the skin barrier lipid structure by inhibiting keratinocyte-degrading enzymes, reducing the breakdown of lipids such as ceramides at the source. Niacinamide has the ability to block the NF-κB signaling pathway, effectively inhibiting the release of inflammatory factors such as IL-6 and TNF-α, and reducing inflammatory responses. Furthermore, niacinamide can regulate melanosome transport, inhibit tyrosinase activity, reduce post-inflammatory hyperpigmentation, and improve uneven skin tone, thus providing both anti-inflammatory benefits and skin barrier repair.

[0035] Grape flower cell extracts are derived from the Merlot and Cabernet Sauvignon grape varieties, which are among the richest in polyphenols (tannins, anthocyanins) in grapes. This gives the extracts strong antioxidant properties, effectively scavenging free radicals in the body and achieving anti-aging effects. Regarding barrier repair, current research shows that grape flower cell extracts can not only increase epidermal moisture retention and reduce transepidermal water loss, improving the skin's physical barrier composed of moisture balance, but also enhance the chemical barrier composed of antimicrobial peptides that control skin bacterial growth by regulating the levels of antimicrobial peptides on the skin surface, promoting barrier repair from different levels and targets.

[0036] This invention utilizes a combination of panthenol, niacinamide, and grape flower cell extract. These components work synergistically to upregulate the expression of barrier-related genes IVL, LOR, and FLG, thereby enhancing skin barrier function and promoting barrier repair. Simultaneously, this composition inhibits the expression of the inflammatory factor IL-6, thus suppressing inflammation, soothing discomfort, and improving inflammatory pigmentation after cosmetic procedures. Finally, this composition was added to a sunscreen formulation at a preferred ratio of 0.3:3:0.1. Human clinical trials have confirmed that, 15 minutes after application, it effectively improves the moisture content of the stratum corneum, transepidermal water loss (TEWL), and the area and degree of redness, providing real-world evidence of the composition's repairing and soothing effects.

[0037] In a second aspect, the present invention provides the application of the composition for post-medical aesthetic repair and soothing according to the first aspect in cosmetics.

[0038] Preferably, the cosmetic includes toner, lotion, cream, mask, serum, or sunscreen.

[0039] Thirdly, the present invention applies the composition to a sunscreen formulation to provide a sunscreen suitable for use after cosmetic procedures, the sunscreen comprising a sunscreen agent, a skin conditioning agent, a pH adjuster, and water; the skin conditioning agent comprising the composition described in the first aspect for repair and soothing after cosmetic procedures.

[0040] The panthenol used in this invention was purchased from Shanghai Qiuyan Trading Co., Ltd.; the nicotinamide was purchased from Shanghai Qiuyan Trading Co., Ltd.; and the grape flower cell extract was purchased from Guangzhou Xincheng Meike Biotechnology Co., Ltd.

[0041] In Examples 1 to 4, Comparative Examples 1 to 7, and Experimental Examples 1 to 4 of the present invention, the raw materials and reagents used can all be purchased from the market.

[0042] The present invention will be further illustrated below with reference to the embodiments: Examples 1 to 4 and Comparative Examples 1 to 7 Table 1. Formulas provided in the embodiments and comparative examples of this application.

[0043]

[0044] Experiment 1: Promoting IVL gene expression

[0045] Test method: 1. Cell Culture Human keratinocytes (Hacat, passage 16) were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) at 37°C in a 5% CO2 incubator. Cells were harvested from the monolayer culture and seeded into 12-well plates at 1.5 × 10⁶ cells per well. 5 Cells were cultured for 6 hours until they adhered to the culture vessel, and then pretreated with serum-free medium for 18 hours.

[0046] 2. Test Grouping

[0047] Table 2. Barrier Repair In Vitro Test Grouping

[0048]

[0049] 3. Administration

[0050] According to the test groups, when the cell seeding rate in the 12-well plates reached 30%–50%, the drugs were administered to the groups, with 3 replicates per group. 1 mL of DMEM medium was added to each well for the blank control and negative control, 1 mL of DMEM medium containing 100 ng / mL LGF-β was added to each well for the positive control, and 1 mL of DMEM medium containing the corresponding test sample was added to each well for the sample group. Except for the blank control, all other groups underwent LPS stimulation. After drug administration, the 12-well plates were incubated for 24 hours.

[0051] 4. RNA extraction and qPCR

[0052] RNA extraction: Total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the instructions. The concentration and purity of RNA were measured using a spectrophotometer to ensure that the OD260 / OD280 ratio was between 1.8 and 2.0.

[0053] Reverse transcription: Take 1 μg of total RNA and use TOYOBO's ReverTra Ace™ qPCR RT Master Mix for reverse transcription to synthesize cDNA.

[0054] qPCR: Using cDNA as a template, qPCR reactions were performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer, 2 μL cDNA template, and 6.4 μL ddH2O. The reaction conditions were: 95℃ pre-denaturation for 30 seconds; 95℃ denaturation for 5 seconds; 60℃ annealing for 30 seconds, for a total of 40 cycles. GAPDH was used as an internal reference gene, and the relative expression level of the IVL gene was calculated using the 2^-ΔΔCt method.

[0055] Table 3 Primers for the target gene

[0056]

[0057] 5. Calculation of Improvement Rate: The improvement rate is calculated using the following formula.

[0058] 6. Statistical Analysis of Results: When comparing groups using the t-test method, the significance level was determined by comparing the NC group with the t-test method. * This means that a p-value < 0.05 indicates that... * A p-value < 0.01 indicates that... ** A p-value < 0.001 indicates that... *** A p-value < 0.0001 indicates that... **** .

[0059] The results are shown in Table 4 and Figure 1 Table 4 shows the relative expression levels and enhancement rates of the target genes provided in Example 1. Figure 1 A bar chart showing the expression levels of the target gene provided in Experiment Example 1.

[0060] Table 4. Expression level and enhancement rate of IVL gene

[0061]

[0062] Table 4 corresponds to Figure 1 The data.

[0063] 7. IVL gene results analysis: In the construction of the stratum corneum barrier, inner lamina protein (IVL) forms an insoluble keratin sheath with laminarin and filaggrin through a transglutaminase-mediated cross-linking reaction, becoming the core functional unit of the physical barrier. This protein complex effectively protects against mechanical irritation, chemical corrosion, and enzymatic damage, and together with the lipid bilayer, maintains the integrity of the stratum corneum barrier. (See Table 4 and...) Figure 1 It can be seen that, compared with the BC group, the IVL expression in the NC group was significantly reduced, indicating that the stimulation conditions of this test were effective; compared with the NC group, the IVL expression in the PC group was significantly increased, indicating that the positive control of this test was effective; compared with the NC group, the IVL expression levels of Comparative Examples 1, 3, 4, 6, and Examples 1, 2, 3, and 4 were significantly increased, with increase rates of 180%, 116%, 85%, 59% and 240%, 145%, 107%, 68%, respectively.

[0064] Furthermore, according to the King's method for calculating synergistic effects:

[0065] Compared to Comparative Examples 1 and 2, the q-value of Example 1 was 1.60 (>1.15). Therefore, the composition provided in Example 1 showed a significant synergistic effect in promoting IVL gene expression levels compared to a single agent. Compared to Comparative Examples 2 and 3, the q-value of Example 2 was 1.32 (>1.15). Therefore, the composition provided in Example 2 showed a significant synergistic effect in promoting IVL gene expression levels compared to a single agent. Compared to Comparative Examples 4 and 5, the q-value of Example 3 was 1.18 (>1.15). Therefore, the composition provided in Example 3 showed a significant synergistic effect in promoting IVL gene expression levels compared to a single agent. In summary, the compositions of the examples can exert a potential barrier repair function by significantly upregulating IVL, an important gene for barrier repair, and are superior to the combination of panthenol and nicotinamide, as well as grape flower cell extract, in terms of the repair mechanism related to this gene level.

[0066] Experimental Example 2: Promoting the Expression of the Barrier Repair Gene LOR

[0067] Test method: 1. Cell Culture Human keratinocytes (Hacat, passage 16) were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) at 37°C in a 5% CO2 incubator. Cells were harvested from the monolayer culture and seeded into 12-well plates at 1.5 × 10⁶ cells per well. 5 Cells were cultured for 6 hours until they adhered to the culture vessel, and then pretreated with serum-free medium for 18 hours.

[0068] 2. Test Grouping

[0069] Table 5. Barrier Repair In Vitro Test Grouping

[0070]

[0071] 3. Administration

[0072] According to the test groups, when the cell seeding rate in the 12-well plates reached 30%–50%, the drugs were administered to the groups, with 3 replicates per group. 1 mL of DMEM medium was added to each well for the blank control and negative control, 1 mL of DMEM medium containing 100 ng / mL LGF-β was added to each well for the positive control, and 1 mL of DMEM medium containing the corresponding test sample was added to each well for the sample group. Except for the blank control, all other groups underwent LPS stimulation. After drug administration, the 12-well plates were incubated for 24 hours.

[0073] 4. RNA extraction and qPCR

[0074] RNA extraction: Total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the instructions. The concentration and purity of RNA were measured using a spectrophotometer to ensure that the OD260 / OD280 ratio was between 1.8 and 2.0.

[0075] Reverse transcription: Take 1 μg of total RNA and use TOYOBO's ReverTra Ace™ qPCR RT Master Mix for reverse transcription to synthesize cDNA.

[0076] qPCR: Using cDNA as a template, qPCR reactions were performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer, 2 μL cDNA template, and 6.4 μL ddH2O. The reaction conditions were: 95℃ pre-denaturation for 30 seconds; 95℃ denaturation for 5 seconds; 60℃ annealing for 30 seconds, for a total of 40 cycles. GAPDH was used as an internal reference gene, and the relative expression level of the LOR gene was calculated using the 2^-ΔΔCt method.

[0077] Table 6 Primers for the target gene

[0078]

[0079] 5. Calculation of Improvement Rate: The improvement rate is calculated using the following formula.

[0080] 6. Statistical Analysis of Results: When comparing groups using the t-test method, the significance level was determined by comparing the NC group with the t-test method. * This means that a p-value < 0.05 indicates that... * A p-value < 0.01 indicates that... ** A p-value < 0.001 indicates that... *** A p-value < 0.0001 indicates that... **** .

[0081] LOR gene results analysis: The results are shown in Table 7 and Figure 2 Table 7 shows the relative expression levels and enhancement rates of the target genes provided in Experiment Example 2. Figure 2 A bar chart showing the expression levels of the target gene provided for Experiment Example 2.

[0082] Table 7 Expression levels and enhancement rates of the LOR gene

[0083]

[0084] Table 7 corresponds to Figure 2 The data.

[0085] LOR (Leydigin of Oleoma), a key structural component of the keratinized capsule, forms an insoluble, tough scaffold around keratinocytes through a transglutaminase-mediated cross-linking reaction. This significantly enhances the mechanical strength and tensile strength of the epidermis, further compensating for the inadequacy of relying solely on moisturizing factors to maintain barrier function. (See Table 7 and...) Figure 2 It can be seen that, compared with the BC group, the LOR expression in the NC group was significantly reduced, indicating that the stimulation conditions of this test were effective; compared with the NC group, the LOR expression in the PC group was significantly increased, indicating that the positive control of this test was effective; compared with the NC group, the LOR expression levels of Comparative Examples 1, 3, and 4, and Examples 1, 2, 3, and 4 were significantly increased, with increase rates of 134%, 85%, and 82% and 187%, 97%, 95%, and 67%, respectively.

[0086] Furthermore, according to the King's method for calculating synergistic effects:

[0087] Compared to Comparative Examples 1 and 2, the q-value of Example 1 was 1.49 (>1.15). Therefore, the composition provided in Example 1 achieved a significant synergistic effect in promoting LOR gene expression levels compared to a single agent. Compared to Comparative Examples 6 and 7, the q-value of Example 4 was 1.27 (>1.15). Therefore, the composition provided in Example 4 achieved a significant synergistic effect in promoting LOR gene expression levels compared to a single agent. In summary, the compositions of the examples can exert a potential barrier repair function by significantly upregulating the barrier repair gene LOR, and are superior to the combination of panthenol and nicotinamide, as well as grape flower cell extract, in the repair mechanism related to this gene level.

[0088] Experiment 3: Promoting the expression of the barrier repair gene FLG

[0089] Test method: 1. Cell Culture Human keratinocytes (Hacat, passage 16) were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) at 37°C in a 5% CO2 incubator. Cells were harvested from the monolayer culture and seeded into 12-well plates at 1.5 × 10⁶ cells per well. 5 Cells were cultured for 6 hours until they adhered to the culture vessel, and then pretreated with serum-free medium for 18 hours.

[0090] 2. Test Grouping

[0091] Table 8. Barrier Repair In Vitro Test Groups

[0092]

[0093] 3. Administration

[0094] According to the test groups, when the cell seeding rate in the 12-well plates reached 30%–50%, the drugs were administered to the groups, with 3 replicates per group. 1 mL of DMEM medium was added to each well for the blank control and negative control, 1 mL of DMEM medium containing 100 ng / mL LGF-β was added to each well for the positive control, and 1 mL of DMEM medium containing the corresponding test sample was added to each well for the sample group. Except for the blank control, all other groups underwent LPS stimulation. After drug administration, the 12-well plates were incubated for 24 hours.

[0095] 4. RNA extraction and qPCR

[0096] RNA extraction: Total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the instructions. The concentration and purity of RNA were measured using a spectrophotometer to ensure that the OD260 / OD280 ratio was between 1.8 and 2.0.

[0097] Reverse transcription: Take 1 μg of total RNA and use TOYOBO's ReverTra Ace™ qPCR RT Master Mix for reverse transcription to synthesize cDNA.

[0098] qPCR: Using cDNA as a template, qPCR reactions were performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer, 2 μL cDNA template, and 6.4 μL ddH2O. The reaction conditions were: 95℃ pre-denaturation for 30 seconds; 95℃ denaturation for 5 seconds; 60℃ annealing for 30 seconds, for a total of 40 cycles. GAPDH was used as an internal reference gene, and the relative expression level of the FLG gene was calculated using the 2^-ΔΔCt method.

[0099] Table 9 Primers for the target gene

[0100]

[0101] 5. Calculation of Improvement Rate: The improvement rate is calculated using the following formula.

[0102] 6. Statistical Analysis of Results: When comparing groups using the t-test method, the significance level was determined by comparing the NC group with the t-test method. * This means that a p-value < 0.05 indicates that... * A p-value < 0.01 indicates that... ** A p-value < 0.001 indicates that... *** A p-value < 0.0001 indicates that... **** .

[0103] FLG gene results analysis: The results are shown in Table 10 and... Figure 3 Table 10 shows the relative expression levels and enhancement rates of the target genes provided in Experiment Example 3. Figure 3 A bar chart showing the expression levels of the target gene provided for Experiment Example 3.

[0104] Table 10 Expression level and enhancement rate of FLG gene

[0105]

[0106] Table 10 corresponds to Figure 3 The data.

[0107] In skin barrier repair, filaggrin (FLG) plays a crucial role by encoding and expressing profilaggrin and its proteolytic products: the natural moisturizing factor (NMF) generated from its degradation maintains stratum corneum hydration and pH balance, while promoting the tight arrangement of keratinocytes, thereby systematically repairing damaged physical and chemical barriers. (See Table 10 and...) Figure 3 It can be seen that, compared with the BC group, the FLG expression in the NC group was significantly reduced, indicating that the stimulation conditions of this test were effective; compared with the NC group, the FLG expression in the PC group was significantly increased, indicating that the positive control of this test was effective; compared with the NC group, the FLG expression levels of Comparative Examples 1, 2, 3, 4, and 6, and Examples 1, 2, 3, and 4 were significantly increased, with increase rates of 109%, 34%, 70%, 56%, 44% and 134%, 78%, 61%, 49%, respectively.

[0108] Furthermore, according to the King's method for calculating synergistic effects:

[0109] Compared to Comparative Examples 1 and 2, the q-value of Example 1 was 1.26 (>1.15). Therefore, the composition provided in Example 1 achieved a significant synergistic effect in promoting FLG gene expression levels compared to single agents. In summary, the composition of the examples can exert its potential barrier repair function by significantly upregulating the barrier repair gene FLG, and it is superior to the combination of panthenol and nicotinamide, as well as grape flower cell extract, in terms of the repair mechanism related to this gene level.

[0110] Therefore, the combination of panthenol, niacinamide, and grape flower cell extract helps to enhance the expression of barrier repair-related genes IVL, LOR, and FLG, and has a synergistic effect. IVL, LOR, and FLG are all key structural proteins that constitute the skin's physical barrier, and their expression levels are often used as important indicators of the integrity of skin barrier function. In the barrier reconstruction stage after cosmetic procedures, the promotion of these barrier genes plays a crucial role in the recovery of barrier function and the reconstruction of barrier integrity. Therefore, the combination of panthenol, niacinamide, and grape flower cell extract exhibits significant repair efficacy.

[0111] Experiment 4: Inhibition of inflammatory factor expression

[0112] Test method: 1. Cell Culture Human keratinocytes (Hacat, passage 16) were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) at 37°C in a 5% CO2 incubator. Cells were harvested from the monolayer culture and seeded into 12-well plates at 1.5 × 10⁶ cells per well.5 Cells were cultured for 6 hours until they adhered to the culture vessel, and then pretreated with serum-free medium for 18 hours.

[0113] 2. Test Grouping

[0114] Table 11 Grouping of in vitro anti-inflammatory test

[0115]

[0116] 3. Administration

[0117] According to the test groups, when the cell seeding rate in the 12-well plates reached 30%–50%, the drugs were administered to the groups, with 3 replicates per group. 1 mL of DMEM medium was added to each well for the blank control and negative control, 1 mL of DMEM medium containing 0.01% asiaticoside was added to each well for the positive control, and 1 mL of DMEM medium containing the corresponding test sample was added to each well for the sample group. Except for the blank group, all other groups underwent LPS stimulation. After drug administration, the 12-well plates were incubated for 24 hours.

[0118] 4. RNA extraction and qPCR

[0119] RNA extraction: Total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the instructions. The concentration and purity of RNA were measured using a spectrophotometer to ensure that the OD260 / OD280 ratio was between 1.8 and 2.0.

[0120] Reverse transcription: Take 1 μg of total RNA and use TOYOBO's ReverTra Ace™ qPCR RT Master Mix for reverse transcription to synthesize cDNA.

[0121] qPCR: Using cDNA as a template, qPCR reactions were performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer, 2 μL cDNA template, and 6.4 μL ddH2O. The reaction conditions were: 95℃ pre-denaturation for 30 seconds; 95℃ denaturation for 5 seconds; 60℃ annealing for 30 seconds, for a total of 40 cycles. GAPDH was used as an internal reference gene, and the relative expression level of the IL-6 gene was calculated using the 2^-ΔΔCt method.

[0122] Table 12 Primers for the target gene

[0123]

[0124] 5. Inhibition rate calculation: The inhibition rate is calculated according to the following formula:

[0125] 6. Statistical Analysis of Results: When comparing groups using the t-test method, the significance level was determined by comparing the NC group with the t-test method. * This means that a p-value < 0.05 indicates that... * A p-value < 0.01 indicates that... ** A p-value < 0.001 indicates that... *** A p-value < 0.0001 indicates that... **** .

[0126] 7. IL-6 gene result analysis

[0127] The results are shown in Table 13 and Figure 4 Table 13 shows the relative expression levels and inhibition rates of the target genes provided in Experiment Example 4. Figure 4 A bar chart showing the expression levels of the target gene provided for Experiment Example 4.

[0128] Table 13 Expression level and inhibition rate of IL-6 gene

[0129]

[0130] Table 13 corresponds to Figure 4 The data.

[0131] Interleukin-6 (IL-6), a key pro-inflammatory cytokine mediating the skin inflammatory cascade, often triggers local immune responses due to its high expression, leading to skin barrier damage and sensitivity phenomena such as redness, swelling, and stinging. Therefore, by specifically inhibiting the expression or release of IL-6, downstream inflammatory signaling pathways can be effectively blocked, thereby achieving anti-inflammatory and soothing effects such as reducing skin erythema and relieving irritation; it also helps to reduce post-inflammatory hyperpigmentation and avoid the risk of post-cosmetic procedures with hyperpigmentation. (See Table 13 and...) Figure 4 It was found that, compared with the BC group, the IL-6 gene expression in the NC group was significantly increased, indicating that the stimulation conditions in this test were effective; compared with the NC group, the IL-6 expression in the PC group was significantly decreased, indicating that the positive control in this test was effective; compared with the NC group, the IL-6 gene expression in Examples 1, 2, 3, and 4 was significantly decreased, with inhibition rates of 69%, 54%, 50%, and 44%, respectively. Therefore, the combination of panthenol, nicotinamide, and grape flower cell extract helps to inhibit the gene expression of the pro-inflammatory factor IL-6 in vitro, thereby exerting a significant anti-inflammatory and soothing effect after cosmetic procedures, and showing strong potential value in relieving postoperative discomfort.

[0132] Experiment 5: Human clinical trials verify repair and soothing effects

[0133] Evaluation sample: A sunscreen containing a composition of panthenol, niacinamide, and grape flower cell extract. Among them: The composition was added to the sunscreen formulation in the proportions of Example 1, with panthenol: niacinamide: grape flower cell extract = 0.3:3.0:0.1.

[0134] Objective: To assess the repair efficacy of an adult female subject with sensitive skin who had experienced tape peeling damage, the test sample was applied once using the prescribed method. Skin moisture and transepidermal water loss were measured in the test sample area and a control area using an instrument probe. Additionally, under normal conditions, the test sample was applied once using the prescribed method. Skin color and redness were assessed on the test sample and control sides using facial image analysis. Furthermore, efficacy was evaluated by a dermatologist to assess the soothing effect of the test sample.

[0135] Evaluation type: randomized, single-blind, controlled.

[0136] Inclusion criteria for participants: female, 24-45 years old, Asian (China), residence in Shanghai, in good health, with no obvious damage or hair on the arm test site that could affect the interpretation of test results, and redness on the skin of the left and right cheeks.

[0137] How to use: 1. Medial forearm: Based on a random table, a marked area of ​​2×2cm was selected on the medial side of one forearm as the evaluation area. 2 Take (2.0±0.1) mg / cm³ 2 Apply the product in a single application, using a latex finger cot for even application; the marked area on the inner side of the other forearm is a blank control area and is not treated in any way.

[0138] 2. Face: According to the randomization table, one side of the face was randomly selected as the evaluation sample side. 0.5g of the product was applied once, and the product was spread evenly with a latex finger cot and gently massaged until absorbed by the skin. The other side of the face was the blank control side and was not treated in any way.

[0139] Evaluation time points: before sample application, immediately after lesion removal (forearm only), and 15 minutes after sample application.

[0140] Evaluation items and instruments: 1. Skin Moisture: Instrument: Corneometer® CM825; Parameter: Skin stratum corneum moisture content (unit: CU); Site: Marked areas on the inner sides of the left and right forearms, and the left and right cheeks; Time points: Before sample application, immediately after detachment of the lesion (forearm only), and 15 minutes after sample application; 2. Transepidermal water loss (TEWL) of the skin: Instrument: Tewameter® TM Hex; Parameter: Transepidermal water loss (TEWL) value (unit: g / h / m³) 2 Location: Marked areas on the inner sides of the left and right forearms, and the left and right cheeks; Time points: before sample application, immediately after lesion removal (forearms only), and 15 minutes after sample application; 3. High-definition digital imaging - skin redness: Instrument: VISIA 7; Location: Left cheek, right cheek; Light source: Standard light, polarized light; Image: VISIA 7 polarized light RBX red side image; Parameters: Image-Pro Plus software analysis of skin redness area; Time points: Before sample application, 15 minutes after sample application; 4. Physician assessment: The dermatologist will assess the redness of the subject's skin; scoring criteria: 0 to 9 rating scale; parameters: rating scale; location: left and right cheeks; time points: before application of sample and 15 minutes after application of sample; Evaluation Results: Thirty-two Asian adult female subjects aged 24 to 45 years underwent a single application of the evaluation sample after tape peeling damage. The stratum corneum moisture content and transepidermal water loss (TEWL) values ​​of the skin in the evaluation sample area and the control area were measured using an instrument probe. The results showed that the evaluation sample had a repairing effect 15 minutes after application. Thirty-two Asian adult female subjects aged 24 to 45 years also underwent a single application of the evaluation sample under normal conditions. The stratum corneum moisture content and transepidermal water loss (TEWL) values ​​of the skin in the evaluation sample area and the control area were measured using an instrument probe. The results showed that after 15 minutes of application, the evaluation sample improved stratum corneum moisture content, TEWL values, and the area and degree of redness, demonstrating a significant immediate effect of barrier repair and redness relief.

[0141] Table 14 Intergroup comparison of forearm test sites and immediate post-ablation injury

[0142]

[0143] Table 15 Comparison between groups after product application on left and right cheek areas

[0144]

[0145] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A repairing and soothing composition, characterized in that, include: Panthenol, nicotinamide, and grape flower cell extract; The mass ratio of the panthenol, the nicotinamide, and the grape flower cell extract is (0.01~5):(0.01~5):(0.00001~0.3).

2. The repairing and soothing composition as described in claim 1, characterized in that, The mass ratio of the panthenol, the nicotinamide, and the grape flower cell extract is (0.01~0.5):(0.01~5):(0.00001~0.3).

3. The repairing and soothing composition as described in claim 2, characterized in that, The mass ratio of the panthenol, the nicotinamide, and the grape flower cell extract is (0.03~0.3):(0.03~3):(0.001~0.1).

4. The repairing and soothing composition as described in claim 3, characterized in that, The mass ratio of the panthenol, the nicotinamide, and the grape flower cell extract is 0.3:3:0.

1.

5. The use of the repairing and soothing composition according to any one of claims 1 to 4 in the preparation of anti-inflammatory, repairing and / or soothing products.

6. The use of the repairing and soothing composition according to any one of claims 1 to 4 in the preparation of a product that upregulates the expression of the IVL gene.

7. The use of the repairing and soothing composition according to any one of claims 1 to 4 in the preparation of products that inhibit and / or reduce the expression of inflammatory factors.

8. The use of the repairing and soothing composition according to any one of claims 1 to 4 in the preparation of products that upregulate the expression of the FLG gene and / or the LOR gene.

9. A product characterized in that, include: The repairing and soothing composition as described in any one of claims 1 to 4.

10. The product as described in claim 9, characterized in that, The products include: personal care products.

Images