Method of building a non-invasive anti-aging skin care system targeting the protection of the dermal collagen network

Abstract

This invention relates to the fields of daily chemical products and skin care, and discloses a method for constructing a non-invasive anti-aging skin care system that targets and protects the dermal collagen network. The method involves obtaining at least three skin layering parameters from dermal echo density, skin elasticity, transepidermal water loss, stratum corneum moisture content, and erythema index, and determining the collagen network risk level accordingly. Based on the collagen network risk level, the method adjusts the outer layer recognition density, response connectivity ratio, and active ingredient loading ratio of dual-gated vesicles in the collagen damage microenvironment. A dual-gated vesicle in the collagen damage microenvironment, comprising a lipid biomimetic membrane layer, a collagen damage recognition layer, a dual-response connectivity layer, and a zoned release core, is prepared. This vesicle is then configured in a stratum corneum hydration flux regulating layer, a collagen affinity response layer, and a surface water-retaining film-forming layer to form a non-invasive anti-aging skin care system. This application improves the matching between the skin care system and the collagen network state through non-invasive grading, selective recognition of damaged collagen, MMP / ROS dual-response release, and layered system configuration.

Description

Technical Field

[0001] This invention relates to the fields of daily chemical products and skin care, specifically to a method for constructing a non-invasive anti-aging skin care system that targets and protects the dermal collagen network. Background Technology

[0002] As skincare needs shift from basic hydration to anti-aging, firming, and barrier maintenance, skincare systems focused on improving dermal collagen have become a key direction in anti-aging product development. Type I and Type III collagen in the dermis constitute the skin's main support network; their arrangement, fiber integrity, and binding to the extracellular matrix affect skin elasticity, plumpness, and the appearance of fine lines. Existing anti-aging skincare products typically attempt to reduce the impact of external oxidative stress, glycation reactions, and matrix metalloproteinase activity on the collagen network, or improve skin aging by adding antioxidants, anti-glycation components, peptides, hyaluronic acid, collagen, retinol derivatives, or plant extracts, or by promoting collagen-related expression.

[0003] In existing technologies, some solutions utilize liposomes, nanoemulsions, exosomes, or other vesicle carriers to encapsulate anti-aging active ingredients, thereby improving the stability of the active ingredients in the formulation and their distribution on the skin surface. These technologies typically focus on the encapsulation efficiency, particle size stability, transdermal permeability, or synergistic effects of active ingredients, thus improving to some extent the problems of easy inactivation, difficulty in uniform dispersion, or unstable user experience in traditional anti-aging compositions. However, in the actual construction of skincare systems, there are individual differences in the aging degree of the dermal collagen network, the state of the stratum corneum barrier, the skin hydration state, and the local oxidative stress state. Fixed-ratio anti-aging formulations are difficult to construct to address different collagen network risk states.

[0004] Furthermore, existing anti-aging skincare systems often focus on enhancing the penetration of active ingredients or increasing the content of collagen-promoting active ingredients, easily overlooking whether these active ingredients can preferentially target areas of loose, denatured, or enzymatically degraded collagen fibers after reaching the skin. Some penetration-enhancing solutions rely on high proportions of alcohols, strong penetration enhancers, or other methods that may affect the continuity of the stratum corneum. While enhancing the entry of active ingredients into the skin, these may also lead to problems such as irritation, barrier fluctuations, or decreased compatibility stability. Existing collagen peptides, polypeptide anti-aging, antioxidant and anti-glycation compositions, and ordinary vesicle carriers typically do not uniformly correlate non-invasive skin layering parameters, selective recognition of damaged collagen, dual-response release of matrix metalloproteinases / reactive oxygen species, and the construction parameters of layered skincare systems. This results in problems such as unclear target sites, unclear release triggering conditions, and insufficient matching of formulation parameters with skin conditions in the direction of collagen network protection. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network, thereby solving the technical problems existing in the prior art.

[0006] The above-mentioned technical objective of the present invention is achieved through the following technical solution: A method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network includes the following steps: S1: Obtain skin layer status parameters, which include at least three of the following: dermal echo density, skin rebound rate, transepidermal water loss, stratum corneum water content, and erythema index, and determine the collagen network risk level based on the skin layer status parameters; S2: Determine the outer layer recognition density, response connectivity ratio, and active ingredient loading ratio of dual-gated vesicles in the collagen damage microenvironment based on the collagen network risk level; S3: Prepare a dual-gated vesicle for collagen damage microenvironment. The dual-gated vesicle for collagen damage microenvironment includes a lipid biomimetic membrane layer, a collagen damage recognition layer, a dual-response connection layer and a partitioned release core. The collagen damage recognition layer is located on the outside of the lipid biomimetic membrane layer, and the dual-response connection layer connects the lipid biomimetic membrane layer and the partitioned release core. S4: The collagen damage recognition layer contains damaged collagen recognition peptides. The amount of collagen damage microenvironment dual-gated vesicles modified with damaged collagen recognition peptides that bind to thermally denatured type I collagen membranes or enzymatically hydrolyzed type I collagen membranes after incubation at 37°C and pH 7.4 for 30 min is 2.0 to 5.0 times the amount that binds to intact type I collagen membranes after incubation for 30 min. S5: The dual-response linker layer contains both matrix metalloproteinase-sensitive linker units and reactive oxygen species-sensitive linker units. The matrix metalloproteinase-sensitive linker units contain peptides that can be cleaved by at least one of MMP-1, MMP-3, and MMP-9. The reactive oxygen species-sensitive linker units contain thioacetyl linker groups. S6: The partitioned release kernel is set as a peripheral release region, an intermediate release region and a kernel release region. The peripheral release region is loaded with oxidative glycation buffer components, the intermediate release region is loaded with network degradation buffer components, and the kernel release region is loaded with collagen arrangement support components. S7: The collagen affinity response layer containing dual-gated vesicles of collagen damage microenvironment, together with the stratum corneum hydration flux regulation layer and the surface water-retaining film layer, are used to construct a non-invasive anti-aging skin care system. The stratum corneum hydration flux regulation layer, collagen affinity response layer and surface water-retaining film layer are arranged in sequence from the skin contact side to the outside.

[0007] Preferably, in step S1, the skin layering state parameters are obtained as follows: dermal echo density is acquired using 20-50MHz high-frequency ultrasound, skin rebound rate is acquired using a skin elasticity tester, transdermal water loss value is acquired using a transdermal water loss tester, stratum corneum water content is acquired using a stratum corneum water content tester, and erythema index is acquired using a skin colorimeter. Before testing, the test skin was equilibrated for 20-30 minutes in an environment with a temperature of 22-25℃ and a relative humidity of 45%-55%. The skin layer state parameters were measured three times consecutively in the same test area and the arithmetic mean was taken.

[0008] Preferably, in step S1, the dermal echo density and skin rebound rate of the test skin detection area are compared with the corresponding parameters of the non-exposed area on the inner side of the forearm of the same test subject. Items with dermal echo density lower than 80% of the non-exposed area of ​​the inner forearm, skin rebound rate lower than 75% of the non-exposed area of ​​the inner forearm, transdermal water loss greater than 15 g / (mh), stratum corneum water content less than 35 a.u., and erythema index greater than 20 are classified as abnormal items. When there are 0 or 1 abnormal items, it is determined to be a Level 1 collagen network risk level; when there are 2 or 3 abnormal items, it is determined to be a Level 2 collagen network risk level; when there are 4 or 5 abnormal items, it is determined to be a Level 3 collagen network risk level. In step S2, when the collagen network risk level is level one, the damaged collagen recognition peptide accounts for 0.5% to 1.5% of the total lipid molar amount in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker unit to reactive oxygen species-sensitive linker unit is 1:1 to 1.5:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 5 to 7:2 to 3:1 to 2. When the collagen network risk level is level 2, the collagen recognition peptide accounts for 1.5% to 3.0% of the total lipid molars in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units is 1.5:1 to 2.5:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 3 to 5:3 to 4:2 to 3. When the collagen network risk level is level three, the damaged collagen recognition peptide accounts for 3.0% to 5.0% of the total lipid molars in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units is 2.5:1 to 4:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 2 to 3:4 to 5:3 to 4.

[0009] Preferably, in step S3, the collagen damage microenvironment dual-gated vesicles are prepared as follows: the lipid biomimetic membrane material is dissolved in ethanol, and the ethanol is removed under reduced pressure to form a lipid film; An aqueous phase containing an oxidative glycation buffer component, a network degradation buffer component, and a collagen arrangement support component was added to a lipid membrane for hydration to obtain a primary emulsion. The primary emulsion was homogenized by microfluidic jetting 3 to 6 times at 60 to 100 MPa, and then granulated using a polycarbonate membrane with a pore size of 100 to 200 nm to obtain basic vesicles. A damaged collagen recognition peptide with a cysteine ​​linker, a matrix metalloproteinase-sensitive linker with a maleimide terminal, and a reactive oxygen species-sensitive linker with a thioketal linker are added to the basic vesicles. This allows the damaged collagen recognition peptide to link to the outer side of the lipid biomimetic membrane, and the matrix metalloproteinase-sensitive linker and the reactive oxygen species-sensitive linker to link between the lipid biomimetic membrane and the partitioned release core.

[0010] Preferably, the lipid biomimetic membrane material comprises, by mass parts, 45-65 parts hydrogenated lecithin, 10-25 parts cholesterol, 1-8 parts ceramide NP, 0.2-3 parts phytosphoin, 0.5-5 parts sodium stearoyl glutamate, and 2-10 parts squalane; The D50 size of the dual-gated vesicles in the collagen damage microenvironment is 50–120 nm, the polydispersity index is no greater than 0.25, and the zeta potential is -5 mV to -30 mV.

[0011] Preferably, in step S4, the damaged collagen recognition peptide is a single-chain collagen mimic peptide, which contains 6 to 10 Gly-Pro-Hyp repeating units arranged in succession. The ends of the single-chain collagen mimic peptide are connected to cysteine ​​linkage groups, and the cysteine ​​linkage groups are connected to the lipid biomimetic membrane layer.

[0012] Preferably, in step S5, the matrix metalloproteinase-sensitive linker unit comprises one of the following: Gly-Pro-Leu-Gly-Val-Arg-Gly peptide, Pro-Leu-Gly-Leu-Ala-Gly peptide, and Gly-Pro-Gln-Gly-Ile-Trp-Gly-Gln peptide. One end of the matrix metalloproteinase-sensitive linker unit is connected to the lipid biomimetic membrane layer, and the other end is connected to the partitioned release core. The reactive oxygen species-sensitive linker includes a thioacetate linker, which is disposed in the outer coating material of the partitioned release core.

[0013] Preferably, in step S6, the oxidative glycation buffer component is composed of ergothioneine, carnosine and ferulic acid glucoside, and the mass ratio of ergothioneine, carnosine and ferulic acid glucoside is 1-3:2-5:1-3. The network degradation buffer component is composed of epigallocatechin gallate, oligomeric proanthocyanidins and low molecular weight sodium hyaluronate, and the mass ratio of epigallocatechin gallate, oligomeric proanthocyanidins and low molecular weight sodium hyaluronate is 3-6:2-4:1-3. The collagen arrangement support component consists of tripeptide-10 citrulline, acetyl tetrapeptide-9, and palmitoyl tripeptide-5, with a mass ratio of 2–4:2–4:1–3.

[0014] Preferably, in step S7, the stratum corneum hydration flux regulating layer comprises, by mass percentage, 0.5%–3.0% betaine, 0.1%–1.0% ectoine, 0.5%–3.0% trehalose, 0.05%–0.5% lactobionic acid, and the balance water, and has a pH of 5.2–5.8 and a viscosity of 500–3000 mPas; The collagen affinity response layer comprises, by mass percentage, 0.05%–2.0% dual-gated vesicles of collagen damage microenvironment, 2.0%–8.0% glycerol, 2.0%–8.0% 1,3-propanediol, and the balance water, with a pH of 5.5–6.2 and a viscosity of 800–5000 mPas; The surface water-retaining film layer comprises, by mass percentage, 0.05%–0.3% low molecular weight sodium hyaluronate, 0.05%–0.5% β-glucan, 0.2%–2.0% pullulan, and the balance water. The dry film thickness after film formation is 2–15 μm, and the film formation time is 3–10 min. The non-invasive anti-aging skincare system is free of azone, dimethyl sulfoxide, and ethanol (greater than 5% by weight).

[0015] Preferably, the cumulative release rate of dual-gated vesicles in the collagen damage microenvironment is no more than 25% after 6 hours in phosphate buffer at pH 7.4; The cumulative release rate was 35%–55% over 6 hours in phosphate buffer containing 50–200 ng / mL matrix metalloproteinases. The cumulative release rate was 55%–80% over 6 hours in phosphate buffer containing 50–200 ng / mL matrix metalloproteinases and 100–500 mol / L hydrogen peroxide. Matrix metalloproteinases are one of MMP-1, MMP-3, and MMP-9.

[0016] In summary, the present invention has the following main beneficial effects: By first acquiring at least three skin layering parameters from dermal echo density, skin rebound rate, transepidermal water loss, stratum corneum moisture content, and erythema index, and comparing the detection area with the non-exposed area of ​​the inner forearm of the same subject, the collagen network risk level is determined. Then, based on the collagen network risk level, the proportions of damaged collagen recognition peptides, matrix metalloproteinase-sensitive linker units (MMPs) and reactive oxygen species (ROS)-sensitive linker units (ROSs), and the loading ratios of the three types of active ingredients are adjusted to achieve a skincare system construction parameter matching the skin collagen network state. Therefore, this application does not use a fixed anti-aging formula to uniformly treat different skin states, but rather transforms non-invasive detection results into a basis for preparing vesicle structures and active ingredient distributions. This gives the system construction process a clear parameter source and adjustment logic, which is beneficial for improving the adaptability and reproducibility of the anti-aging skincare system under different skin states.

[0017] By setting a lipid biomimetic membrane layer, a collagen damage recognition layer, a dual-response linker layer, and a regional release core within dual-gated vesicles in the collagen damage microenvironment, and by ensuring that the damaged collagen recognition peptides in the collagen damage recognition layer bind more to thermally denatured or enzymatically hydrolyzed type I collagen membranes than to intact type I collagen membranes, and by including matrix metalloproteinase-sensitive linker units and reactive oxygen species-sensitive linker units in the dual-response linker layer, selective recognition and responsive release from the collagen damage microenvironment are achieved. Compared to ordinary liposomes, exosome-like carriers, or single-peptide anti-aging combinations, the technical focus of this application is not simply to increase the active ingredient loading or penetration enhancement, but rather to utilize damaged collagen recognition, MMP response, and ROS response to form a continuous vesicle release pathway. This allows the release of active ingredients to correspond to the microenvironmental characteristics such as loose collagen network, enzymatic hydrolysis, and oxidative stress, thereby improving the technical stability of this skincare system, which distinguishes it from conventional anti-aging compound solutions.

[0018] By setting the regional release core as a peripheral release zone, an intermediate release zone, and a core release zone, the oxidative glycation buffer component, the network degradation buffer component, and the collagen arrangement support component are placed in different release positions. Furthermore, dual-gated vesicles representing the collagen damage microenvironment are configured within the stratum corneum hydration flux regulating layer, the collagen affinity response layer, and the surface water-retaining film-forming layer. This achieves the effect of constructing a non-invasive anti-aging skincare system without using microneedles, injections, laser perforation, strong penetration enhancers, or high proportions of ethanol. The stratum corneum hydration flux regulating layer maintains the stratum corneum's hydration state; the collagen affinity response layer carries the dual-gated vesicles; and the surface water-retaining film-forming layer prolongs the surface hydration diffusion time. This allows the three layers to work together in terms of composition, pH, viscosity, film-forming time, and dry film thickness, thus avoiding reliance on strong penetration enhancers or highly irritating active ingredients to increase the intensity of action and improving the system's feasibility in daily skincare scenarios. Attached Figure Description

[0019] Figure 1This is a flowchart of the method of the present invention. Detailed Implementation

[0020] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Example 1 refer to Figure 1 A method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network includes the following steps: S1: Obtain skin layer status parameters, which include at least three of the following: dermal echo density, skin rebound rate, transepidermal water loss, stratum corneum water content, and erythema index, and determine the collagen network risk level based on the skin layer status parameters; S2: Determine the outer layer recognition density, response connectivity ratio, and active ingredient loading ratio of dual-gated vesicles in the collagen damage microenvironment based on the collagen network risk level; S3: Prepare a dual-gated vesicle for collagen damage microenvironment. The dual-gated vesicle for collagen damage microenvironment includes a lipid biomimetic membrane layer, a collagen damage recognition layer, a dual-response connection layer and a partitioned release core. The collagen damage recognition layer is located on the outside of the lipid biomimetic membrane layer, and the dual-response connection layer connects the lipid biomimetic membrane layer and the partitioned release core. S4: The collagen damage recognition layer contains damaged collagen recognition peptides. The amount of collagen damage microenvironment dual-gated vesicles modified with damaged collagen recognition peptides that bind to thermally denatured type I collagen membranes or enzymatically hydrolyzed type I collagen membranes after incubation at 37°C and pH 7.4 for 30 min is 2.0 to 5.0 times the amount that binds to intact type I collagen membranes after incubation for 30 min. S5: The dual-response linker layer contains both matrix metalloproteinase-sensitive linker units and reactive oxygen species-sensitive linker units. The matrix metalloproteinase-sensitive linker units contain peptides that can be cleaved by at least one of MMP-1, MMP-3, and MMP-9. The reactive oxygen species-sensitive linker units contain thioacetyl linker groups. S6: The partitioned release kernel is set as a peripheral release region, an intermediate release region and a kernel release region. The peripheral release region is loaded with oxidative glycation buffer components, the intermediate release region is loaded with network degradation buffer components, and the kernel release region is loaded with collagen arrangement support components. S7: The collagen affinity response layer containing dual-gated vesicles of collagen damage microenvironment, together with the stratum corneum hydration flux regulation layer and the surface water-retaining film layer, are used to construct a non-invasive anti-aging skin care system. The stratum corneum hydration flux regulation layer, collagen affinity response layer and surface water-retaining film layer are arranged in sequence from the skin contact side to the outside.

[0022] This method does not involve the conventional compounding of antioxidants, peptides, hyaluronic acid, or liposomes. Instead, it first determines the risk level of the collagen network through non-invasive detection, and then adjusts the outer layer recognition density, response connectivity ratio, and active ingredient loading ratio of dual-gated vesicles in the collagen damage microenvironment according to the collagen network risk level. Subsequently, the dual-gated vesicles in the collagen damage microenvironment are configured in the stratum corneum hydration flux regulation layer, the collagen affinity response layer, and the surface water-retaining film-forming layer, so that a continuous formulation construction logic is formed between detection grading, vesicle structure, active ingredient distribution, and the configuration of the three-layer system.

[0023] In this embodiment, "non-invasive" refers to the absence of microneedling, injection, laser perforation, radiofrequency ablation, dermabrasion, or other treatments that disrupt the continuity of the stratum corneum. Collagen damage microrings refer to a state in the skin where at least one of the following exists: thermal denaturation of collagen fibers, enzymatic loosening of collagen fibers, increased activity of matrix metalloproteinases, or increased levels of reactive oxygen species. The collagen damage recognition layer refers to a surface layer located on the outer side of the lipid biomimetic membrane layer and containing damaged collagen recognition peptides. The dual-response connection layer refers to a connection structure containing both matrix metalloproteinase-sensitive connection units and reactive oxygen species-sensitive connection units. The partitioned release core refers to a peripheral release zone, an intermediate release zone, and a core release zone formed within the same collagen damage microenvironment's dual-gated vesicles, based on the order of addition of different active ingredients, their complexation state, differences in hydrophilicity, and the position of the coating material.

[0024] During implementation, skin layering parameters are first obtained. The test area can be the cheek, forehead, outer periorbital area, or other skin areas requiring an anti-aging skincare system; the control area is the unexposed inner forearm of the same subject. Using the unexposed inner forearm of the same subject as a control is to reduce the influence of differences in age, skin type, genetic background, and lifestyle on the test results. This control method is only used for the process grading of the skincare system and is not used as a diagnostic criterion for diseases.

[0025] Before testing, the test skin was equilibrated for 20-30 minutes in an environment with a temperature of 22-25℃ and a relative humidity of 45%-55%. No skincare products were applied to the test area within 2 hours prior to testing. Dermal echo density was acquired using 20-50MHz high-frequency ultrasound; skin elasticity was measured using a skin elasticity tester; transdermal water loss was measured using a transdermal water loss tester; stratum corneum moisture content was measured using a stratum corneum moisture content tester; and erythema index was measured using a skin colorimeter. Each skin layer condition parameter was measured three times consecutively in the same test area, and the arithmetic mean was taken. When the relative standard deviation of the three measurements exceeded 10%, the skin was allowed to reequilibrate for 5 minutes before measurement.

[0026] The risk level of collagen networks is determined as follows: ; ; ; in, This represents the normalized value of the dermal echo density; This indicates the dermal echo density of the tested skin area; This refers to the dermal echo density of the non-exposed area on the inner forearm of the same subject. This represents the normalized value of skin elasticity. This indicates the skin rebound rate of the tested skin area; This indicates the skin rebound rate of the non-exposed area on the inner forearm of the same subject; This represents the transepidermal water loss value, expressed in g / (mh). This indicates the water content of the stratum corneum, measured in au. Indicates the erythema index; This is an indicator function that takes the value 1 when the condition inside the parentheses is true and 0 when the condition inside the parentheses is false. This indicates the number of abnormal items, with a value ranging from 0 to 5.

[0027] When the number of abnormal items When there are 0 or 1 abnormal items, it is determined to be a Level 1 collagen network risk level; when the number of abnormal items is 0... When there are 2 or 3 abnormal items, it is determined to be a level 2 collagen network risk level; when the number of abnormal items is... When there are 4 or 5 items, it is determined to be a level 3 collagen network risk level. The above level output results are used to select the proportion of damaged collagen recognition peptides in the total lipid molar amount, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units, and the mass ratio of oxidative glycation buffer components, network degradation buffer components, and collagen arrangement support components.

[0028] The above thresholds are formulation grading thresholds in the skincare system construction process and are not medical diagnostic thresholds. A dermal echo density lower than 80% of the non-exposed area on the inner forearm of the same subject and a skin rebound rate lower than 75% of the non-exposed area on the inner forearm of the same subject indicate that the tested area has shown a stably distinguishable decrease in dermal echo and mechanical recovery ability relative to its own non-exposed area. A transdermal water loss greater than 15 g / (mh), a stratum corneum water content less than 35 a.u., and an erythema index greater than 20 are used to supplement and limit the collagen network risk level based on barrier status, hydration status, and skin surface reaction status. These five parameters collectively serve as the basis for selecting subsequent vesicle construction parameters, avoiding formulation determination solely based on subjective wrinkle observation.

[0029] The outer layer recognition density, response linkage ratio, and active ingredient loading ratio of dual-gated vesicles in the collagen damage microenvironment were determined based on the collagen network risk level. When the collagen network risk level was Level 1, the damaged collagen recognition peptide accounted for 0.5%–1.5% of the total molar amount of lipids in the biomimetic lipid membrane layer; the molar ratio of matrix metalloproteinase-sensitive linkers to reactive oxygen species-sensitive linkers was 1:1–1.5:1; and the mass ratio of oxidative glycation buffer components, network degradation buffer components, and collagen alignment support components was 5–7:2–3:1–2. When the collagen network risk level was Level 2, the damaged collagen recognition peptide accounted for 1.5%–3.0% of the total molar amount of lipids in the biomimetic lipid membrane layer; the molar ratio of matrix metalloproteinase-sensitive linkers to reactive oxygen species-sensitive linkers was 1.5:1–2.5:1; and the mass ratio of oxidative glycation buffer components, network degradation buffer components, and collagen alignment support components was 3–5:3–4:2–3. When the collagen network risk level is level three, the damaged collagen recognition peptide accounts for 3.0% to 5.0% of the total lipid molars in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units is 2.5:1 to 4:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 2 to 3:4 to 5:3 to 4.

[0030] The above-mentioned ratio ranges were determined based on vesicle dispersion stability, the amount of damaged collagen bound, and the dual-response release rate. When the proportion of damaged collagen recognition peptides is too low, the selective binding of vesicles to thermally denatured or enzymatically hydrolyzed type I collagen membranes is insufficient; when the proportion is too high, vesicle surface interactions are enhanced, easily leading to increased particle size and aggregation. The ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units is used to regulate the relationship between enzyme-responsive release and reactive oxygen species-responsive release. The ratios of the three types of active substances are gradually adjusted from primarily oxidative glycation buffering to a balance between network degradation buffering and collagen alignment support, based on the collagen network risk level.

[0031] When preparing dual-gated vesicles for collagen damage microenvironments, the lipid biomimetic membrane material comprises, by weight, 45–65 parts hydrogenated lecithin, 10–25 parts cholesterol, 1–8 parts ceramide NP, 0.2–3 parts phytosphoprotein, 0.5–5 parts sodium stearoyl glutamate, and 2–10 parts squalane. To provide linkage sites for collagen recognition peptides in damaged collagen, 0.1–2 parts of a phospholipid derivative with maleimide groups can be added to the lipid biomimetic membrane material. This phospholipid derivative serves as a linkage aid and does not alter the main composition of the lipid biomimetic membrane.

[0032] The damaged collagen recognition peptide is a single-chain collagen mimic peptide containing 6–10 consecutively arranged Gly-Pro-Hyp repeating units, each terminally linked with a cysteine ​​linker. The cysteine ​​linker reacts with maleimide groups on the surface of the lipid biomimetic membrane via a thiol-maleimide addition reaction, attaching the damaged collagen recognition peptide to the outer side of the membrane. The number of Gly-Pro-Hyp repeating units, set at 6–10, is a range determined by both collagen-related structure recognition and vesicle stability after peptide grafting. With fewer than 6 repeating units, the binding amount to the damaged collagen membrane is difficult to consistently reach more than 2.0 times that of the intact type I collagen membrane; with more than 10 repeating units, the peptide chain length increases, easily leading to vesicle surface entanglement and increased particle size.

[0033] The matrix metalloproteinase-sensitive linker comprises one of the following peptides: Gly-Pro-Leu-Gly-Val-Arg-Gly, Pro-Leu-Gly-Leu-Ala-Gly, and Gly-Pro-Gln-Gly-Ile-Trp-Gly-Gln. One end of the matrix metalloproteinase-sensitive linker is connected to a lipid biomimetic membrane layer, and the other end is connected to a partitioned release core. In practice, one end of the matrix metalloproteinase-sensitive linker can be connected to a cysteine ​​or amino terminal group for reaction with maleimide or carboxyl-activated terminal groups on the lipid biomimetic membrane layer; the other end can be connected to a hydrophobic anchoring group or an amino-modified hyaluronic acid fragment for binding to the outer coating material of the partitioned release core.

[0034] After preparation, the matrix metalloproteinase-sensitive linker can be verified by cleavage using phosphate buffer containing MMP-1, MMP-3, or MMP-9. The linker product containing the matrix metalloproteinase-sensitive linker is incubated with 50–200 ng / mL of the corresponding matrix metalloproteinase at 37°C for 2–6 h. Peptide fragmentation products are detected by high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC-MS). When the corresponding fragmentation product is detected and no identical fragmentation peak appears in the blank phosphate buffer, the linker is confirmed to meet the requirement of being cleaved by at least one of MMP-1, MMP-3, and MMP-9.

[0035] The reactive oxygen species (ROS)-sensitive linker contains a thioketal linker. In implementation, an amphiphilic molecule containing a thioketal linker can be used as part of the outer coating material of the partitioned release core, with the thioketal linker positioned within the outer coating material of the partitioned release core. The amphiphilic molecule containing the thioketal linker can be prepared by amidation reaction of thioketal dicarboxylic acid with aminated lipids or aminated polyethylene glycol lipids, or by using commercially available raw materials with confirmed purity or commissioned synthesis. The thioketal linker undergoes breakage or polarity change in an aqueous environment containing ROS, reducing the compactness of the outer coating material of the partitioned release core. The matrix metalloproteinase-sensitive linker and the ROS-sensitive linker regulate vesicle release from the enzymatic cleavage channel and oxidative stress channel, respectively, and are not simply a mixture of two response materials.

[0036] In the specific preparation process, the lipid biomimetic membrane material is dissolved in ethanol to obtain a lipid phase. The amount of ethanol used is 5 to 20 times the total mass of the lipid biomimetic membrane material, ensuring complete dissolution. The lipid phase is placed in a rotary evaporator and the ethanol is removed under reduced pressure at 35–45°C to form a uniform lipid film on the inner wall of the container. Vacuuming continues for 30–60 minutes to reduce the residual ethanol content.

[0037] A core phase, an intermediate phase, and a peripheral phase were prepared separately. The core phase contained a collagen alignment support component and 0.02%–0.20% low-molecular-weight sodium hyaluronate. The collagen alignment support component consisted of tripeptide-10 citrulline, acetyl tetrapeptide-9, and palmitoyl tripeptide-5, with a mass ratio of 2–4:2–4:1–3. The intermediate phase contained a network degradation buffer component, which consisted of epigallocatechin gallate, oligomeric proanthocyanidins, and low-molecular-weight sodium hyaluronate, with a mass ratio of 3–6:2–4:1–3. The peripheral phase contains an oxidative glycation buffer component; the oxidative glycation buffer component consists of ergothioneine, carnosine and ferulic acid glucoside, with a mass ratio of ergothioneine, carnosine and ferulic acid glucoside of 1–3:2–5:1–3.

[0038] The core phase was added to a lipid membrane and hydrated at 35–45 °C for 20–40 min to obtain a preliminary hydrated solution. An intermediate phase was then added, and hydration continued for 10–20 min to distribute the complexed dispersion structure formed by epigallocatechin gallate, oligomeric proanthocyanidins, and low-molecular-weight sodium hyaluronate on the outer side of the core phase. The resulting system was subjected to microfluidic homogenization at 60–100 MPa 3–6 times, followed by granulation using a polycarbonate membrane with a pore size of 100–200 nm to obtain basic vesicles. After cooling the basic vesicles to 20–25 °C, the peripheral phase was added, and the mixture was stirred at 100–300 rpm for 20–40 min under light-protected conditions to distribute the oxidative glycation buffer components in the peripheral hydration layer of the basic vesicles.

[0039] Subsequently, a collagen-recognizing peptide with a cysteine ​​linker, a matrix metalloproteinase-sensitive linker with a maleimide terminal, and a reactive oxygen species-sensitive linker with a thioacetyl linker were added to the basic vesicles. The reaction was carried out at pH 6.8–7.2 for 1–4 h, which allowed the collagen-recognizing peptide to link to the outer side of the lipid biomimetic membrane, and the matrix metalloproteinase-sensitive linker and the reactive oxygen species-sensitive linker to link between the lipid biomimetic membrane and the partitioned release core, resulting in a dual-gated vesicle with a collagen damage microenvironment.

[0040] The prepared collagen-damaged microenvironment dual-gated vesicles were analyzed by dynamic light scattering. The D50 particle size ranged from 50 to 120 nm, the polydispersity index (PDI) was no greater than 0.25, and the Zeta potential ranged from -5 mV to -30 mV. The D50 particle size range of 50–120 nm was determined based on the dispersion stability of the vesicles after membrane granulation and the spreadability in the collagen affinity response layer. When the D50 particle size is less than 50 nm, higher requirements are placed on homogenization and membrane granulation conditions, and the encapsulation efficiency and batch-to-batch stability are prone to fluctuations. When the D50 particle size is greater than 120 nm, the dispersion stability of the vesicles in the collagen affinity response layer decreases. The PDI of no more than 0.25 is used to limit the uniformity of vesicle particle size distribution. The Zeta potential of -5 mV to -30 mV is used to limit the dispersion state of the vesicles in the aqueous system, avoiding aggregation due to excessively low absolute potential values ​​and avoiding the impact of excessively high absolute potential values ​​on the compatibility stability of the three-layer system.

[0041] To confirm the selective binding capacity of the damaged collagen recognition layer, binding tests were conducted using intact type I collagen membranes, heat-denatured type I collagen membranes, and enzymatically hydrolyzed type I collagen membranes. Intact type I collagen membranes were obtained by forming a film from a type I collagen solution under neutral conditions and then drying it. Heat-denatured type I collagen membranes were obtained by treating intact type I collagen membranes at 55–65°C for 10–30 min. Enzymatically hydrolyzed type I collagen membranes were obtained by treating intact type I collagen membranes with a low concentration of collagenase or MMP followed by thorough washing. Fluorescently labeled dual-gated vesicles of the damaged collagen microenvironment were incubated with the above collagen membranes at 37°C and pH 7.4 for 30 min. After washing away unbound vesicles, the fluorescence intensity or residual vesicle amount on the collagen membrane surface was measured.

[0042] The selective binding factor of damaged collagen is calculated using the following formula: ; in, Indicates the selective binding factor of damaged collagen; This indicates the amount of binding between dual-gated vesicles in the collagen damage microenvironment and thermally denatured type I collagen membranes or enzymatically hydrolyzed type I collagen membranes after incubation for 30 min. This indicates the amount of binding between dual-gated vesicles in the collagen-damaged microenvironment and the intact type I collagen membrane after 30 minutes of incubation. The binding amount can be expressed as fluorescence intensity, residual vesicles per unit area, or a calibrated fluorescence signal conversion value; the same measurement method is used in the same batch of tests.

[0043] when When the value is 2.0–5.0, the dual-gated vesicles in the collagen damage microenvironment are considered to meet the requirements for selective recognition of damaged collagen. This range represents the process balance between selective recognition of damaged collagen and vesicle dispersion stability. When the value is less than 2.0, the collagen damage recognition layer lacks selectivity for thermally denatured type I collagen membranes or enzymatically hydrolyzed type I collagen membranes, making it difficult to distinguish it from ordinary collagen affinity carriers; when When the value is greater than 5.0, it is usually necessary to increase the grafting density of the damaged collagen recognition peptide. However, excessively high grafting density can enhance vesicle surface interactions, increasing the risk of particle size enlargement and aggregation during storage. Therefore, this embodiment will... The control range is 2.0 to 5.0.

[0044] To confirm the release characteristics of the dual-response connecting layer, three release media were used to detect the 6-hour cumulative release rate of dual-gated vesicles in the collagen-damaged microenvironment. The first release medium was a phosphate-buffered saline (PBS) at pH 7.4; the second release medium was a PBS at pH 7.4 containing 50–200 ng / mL matrix metalloproteinases (MMP-1, MMP-3, and MMP-9); and the third release medium was a PBS at pH 7.4 containing both 50–200 ng / mL MMP and 100–500 mol / L hydrogen peroxide. The dual-gated vesicles in the collagen-damaged microenvironment were placed in dialysis bags and released by shaking at 37°C. Samples were taken at 0.5 h, 1 h, 2 h, 4 h, and 6 h to detect the content of marker active substances in the release solution. Ergothionein in the peripheral release region was used as the peripheral release marker, epigallocatechin gallate in the intermediate release region was used as the intermediate release marker, and tripeptide-10 citrulline in the core release region was used as the core release marker.

[0045] The 6-hour cumulative release rate was calculated as the percentage of the cumulative release over 6 hours relative to the initial load. The prepared collagen-damaged microenvironment dual-gated vesicles met the following release conditions: a 6-hour cumulative release rate of no more than 25% in phosphate buffer at pH 7.4; a 6-hour cumulative release rate of 35%–55% in phosphate buffer containing 50–200 ng / mL matrix metalloproteinases; and a 6-hour cumulative release rate of 55%–80% in phosphate buffer containing both 50–200 ng / mL matrix metalloproteinases and 100–500 mol / L hydrogen peroxide. These release rate ranges are used to characterize the gated release state of the dual-response connective layer and are not used as indicators for human efficacy evaluation. A cumulative release rate of no more than 25% over 6 hours in ordinary phosphate buffer indicates that rapid leakage of the vesicles did not occur. A cumulative release rate of 35%–55% over 6 hours under conditions containing matrix metalloproteinases (MMPs) indicates that the release channel partially opens after the MMP-sensitive linker unit is cleaved or loosened. A cumulative release rate of 55%–80% over 6 hours under conditions containing both MMPs and hydrogen peroxide indicates that the permeability of the outer coating material of the nucleus is further improved when the MMP-sensitive linker unit and the reactive oxygen species (ROS)-sensitive linker unit work together. These ranges were determined through control release tests on ordinary liposomes, single MMP-responsive vesicles, single ROS-responsive vesicles, and dual-gated vesicles in the collagen-damaged microenvironment of this embodiment, and were used as quality control standards after vesicle preparation.

[0046] After completing the preparation of dual-gated vesicles in the collagen-damaged microenvironment, a non-invasive anti-aging skincare system was constructed. This system comprises a stratum corneum hydration flux regulating layer, a collagen affinity response layer, and a surface water-retaining film-forming layer, arranged sequentially from the skin contact side to the outer side. This sequential arrangement can be achieved through three dosage forms used in stages, or by storing them in a single packaging system with separate compartments and applying them sequentially during use. The three-layer system is preferably stored separately to prevent the vesicles in the collagen affinity response layer from being continuously immersed in the high-viscosity system of the surface water-retaining film-forming layer, which could affect particle size stability.

[0047] The stratum corneum hydration flux regulating layer comprises, by mass percentage, 0.5%–3.0% betaine, 0.1%–1.0% ectoine, 0.5%–3.0% trehalose, 0.05%–0.5% lactobionic acid, and the balance water, with a pH of 5.2–5.8 and a viscosity of 500–3000 mPas. The amount of lactobionic acid in this layer is controlled at 0.05%–0.5%, and it is not used as an acidic active agent for exfoliating the stratum corneum, but rather works in conjunction with betaine, ectoine, and trehalose to regulate the hydration state of the stratum corneum.

[0048] The collagen affinity-responsive layer comprises, by mass percentage, 0.05%–2.0% dual-gated vesicles representing collagen damage microenvironments, 2.0%–8.0% glycerol, 2.0%–8.0% 1,3-propanediol, and the balance water, with a pH of 5.5–6.2 and a viscosity of 800–5000 mPas. In preparation, glycerol, 1,3-propanediol, and water are first mixed thoroughly. Then, the dual-gated vesicles representing collagen damage microenvironments are slowly added at 20–25°C, and the mixture is stirred at low speed until homogeneous. The preferred stirring speed is 100–300 rpm to minimize vesicle rupture or particle size increase.

[0049] The surface water-retaining film layer comprises, by mass percentage, 0.05%–0.3% low molecular weight sodium hyaluronate, 0.05%–0.5% β-glucan, 0.2%–2.0% pullulan, and the balance water. The dry film thickness after film formation is 2–15 μm, and the film formation time is 3–10 min. This layer forms a continuous hydration film outside the collagen affinity-responsive layer, avoiding the use of excessively sealing waxy thick films.

[0050] The described non-invasive anti-aging skincare system is free of azone, dimethyl sulfoxide, and ethanol (greater than 5% by mass). This limitation is a system composition limitation, used to distinguish it from systems that rely on strong penetration enhancers or high proportions of volatile alcohols to improve penetration. Ethanol content can be confirmed by gas chromatography, or by combining formulation dosage and residual solvent detection. When ethanol is used only as a volatile solvent in the lipid film preparation process, the ethanol mass percentage in the final non-invasive anti-aging skincare system is controlled to be no greater than 5% after ethanol removal under reduced pressure.

[0051] Viscosity was measured using a rotational viscometer at 25°C. Before testing, the sample was allowed to stand for 24 hours to remove air bubbles. The rotor model and speed were selected based on the sample's viscosity range, ensuring the instrument reading was within 20%–80% of its range. Film formation time was determined by uniformly coating a surface water-retaining film layer at a rate of 2 mg / cm² onto an inert glass plate or artificial skin membrane surface, and recording the time it took for the surface to transition from a wet state to a continuous, non-flowing film state in an environment with a temperature of 22–25°C and a relative humidity of 45%–55%. Dry film thickness was calculated by converting the wet film coating amount and solid content, and confirmed by sampling with a film thickness gauge. The dry film thickness, 2–15 μm, refers to the average thickness after the surface water-retaining film layer has fully formed.

[0052] In one specific embodiment, a non-invasive anti-aging skincare system was constructed based on the secondary collagen network risk level. Test results showed that the dermal echo density of the tested skin area relative to the non-exposed area of ​​the inner forearm was less than 80%, the skin rebound rate was less than 75%, the stratum corneum water content was less than 35 a.u., and the transepidermal water dispersion value and erythema index did not meet the abnormal standards. Therefore, the number of abnormal items was 3, and the system was determined to be at the secondary collagen network risk level. Based on the secondary collagen network risk level, the proportion of damaged collagen recognition peptides in the total lipid molar amount in the lipid biomimetic membrane was set to 2.2%, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units was set to 2:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component was set to 4:3.5:2.5.

[0053] In this specific embodiment, the lipid biomimetic membrane material comprises, by mass, 55 parts hydrogenated lecithin, 18 parts cholesterol, 4 parts ceramide NP, 1 part phytosphingosine, 2 parts sodium stearoyl glutamate, and 6 parts squalane, with 0.8 parts of a phospholipid derivative containing maleimide groups added. The above lipid biomimetic membrane material is dissolved in ethanol and subjected to reduced pressure to form a lipid film. A core phase containing tripeptide-10 citrulline, acetyl tetrapeptide-9, palmitoyl tripeptide-5, and low-molecular-weight sodium hyaluronate is added to the lipid film for hydration. An intermediate phase containing epigallocatechin gallate, oligomeric proanthocyanidins, and low-molecular-weight sodium hyaluronate is then added for further hydration. The mixture is homogenized four times using 80 MPa microfluidic jets and granulated using a 100 nm polycarbonate membrane to obtain basic vesicles. Subsequently, a peripheral phase containing ergothioneine, carnosine, and ferulic acid glucoside was added for post-loading. Then, a collagen-recognizing peptide with a cysteine ​​linker, a matrix metalloproteinase-sensitive linker with a maleimide terminal group, and a reactive oxygen species-sensitive linker with a thioketal linker were added. The reaction was carried out at pH 7.0 for 2 hours to obtain dual-gated vesicles in a collagen-damaged microenvironment. The D50 particle size of these dual-gated vesicles in the collagen-damaged microenvironment was controlled within the range of 50–120 nm, the polydispersity index was not greater than 0.25, and the Zeta potential was -5 mV to -30 mV.

[0054] In this specific embodiment, the stratum corneum hydration flux regulating layer comprises, by mass percentage, 1.5% betaine, 0.5% ectoine, 1.5% trehalose, 0.2% lactobionic acid, and the balance water, with the pH adjusted to 5.5 and the viscosity controlled between 500 and 3000 mPas. The collagen affinity responsive layer comprises, by mass percentage, 0.8% dual-gated vesicles of collagen damage microenvironment, 5.0% glycerol, 5.0% 1,3-propanediol, and the balance water, with the pH adjusted to 5.8 and the viscosity controlled between 800 and 5000 mPas. The surface water-retaining film-forming layer comprises, by mass percentage, 0.15% low molecular weight sodium hyaluronate, 0.2% β-glucan, 1.0% pullulan, and the balance water, with the film-forming time controlled between 3 and 10 minutes, and the dry film thickness after film formation between 2 and 15 μm. The three-layer system was prepared and stored separately. When used, the stratum corneum hydration flux regulating layer, collagen affinity response layer and surface water-retaining film layer were applied in sequence.

[0055] To illustrate the structural and process differences of this embodiment, the following comparison methods can be set up. Control 1 is a conventional liposome containing only a lipid biomimetic membrane layer and the same active ingredient, but without damaged collagen recognition peptides, matrix metalloproteinase-sensitive linker units, or reactive oxygen species (ROS)-sensitive linker units. Control 2 is a non-damaged collagen recognition peptide vesicle containing a lipid biomimetic membrane layer, a dual-response linker layer, and a partitioned release core, but without a collagen damage recognition layer. Control 3 is a single MMP-responsive vesicle containing damaged collagen recognition peptides and MMP-sensitive linker units, but without ROS-sensitive linker units. Control 4 is a single ROS-responsive vesicle containing damaged collagen recognition peptides and ROS-sensitive linker units, but without MMP-sensitive linker units. Through the above comparisons, it can be verified that the difference in this embodiment lies not in the encapsulation of anti-aging active ingredients by conventional liposomes, nor in the presence of a single collagen peptide, a single MMP response, or a single ROS response, but in the synergistic construction relationship between non-invasive detection grading, selective recognition of damaged collagen, MMP / ROS dual-response linker, and partitioned release core.

[0056] Compared to conventional exosome or exosome-like anti-aging systems, this embodiment does not use exosome morphology as the core technology. Instead, it controls the binding amount of vesicles to thermally denatured or enzymatically hydrolyzed type I collagen membranes relative to intact type I collagen membranes to be 2.0–5.0 times, and defines the release pathway through an MMP / ROS dual-response connecting layer. Compared to conventional collagen expression-promoting compositions, this embodiment does not use peptides, polyphenols, hyaluronic acid, or antioxidants as fixed compound ingredients, but rather configures them separately in the peripheral release zone, intermediate release zone, and core release zone, and adjusts the proportion of the three types of active ingredients according to the risk level of the collagen network. Compared to single MMP-responsive or single ROS-responsive delivery materials, this embodiment is limited to use in non-invasive skin care systems, and is configured sequentially through a stratum corneum hydration flux regulating layer, a collagen affinity response layer, and a surface water-retaining film-forming layer, avoiding reliance on microneedles, injections, or strong penetration enhancers for delivery.

[0057] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network, characterized in that, Includes the following steps: S1: Obtain skin layer status parameters, which include at least three of the following: dermal echo density, skin rebound rate, transepidermal water loss, stratum corneum water content, and erythema index, and determine the collagen network risk level based on the skin layer status parameters; S2: Determine the outer layer recognition density, response connectivity ratio, and active ingredient loading ratio of dual-gated vesicles in the collagen damage microenvironment based on the collagen network risk level; S3: Prepare a dual-gated vesicle for collagen damage microenvironment. The dual-gated vesicle for collagen damage microenvironment includes a lipid biomimetic membrane layer, a collagen damage recognition layer, a dual-response connection layer and a partitioned release core. The collagen damage recognition layer is located on the outside of the lipid biomimetic membrane layer, and the dual-response connection layer connects the lipid biomimetic membrane layer and the partitioned release core. S4: The collagen damage recognition layer contains damaged collagen recognition peptides. The amount of collagen damage microenvironment dual-gated vesicles modified with damaged collagen recognition peptides that bind to thermally denatured type I collagen membranes or enzymatically hydrolyzed type I collagen membranes after incubation at 37°C and pH 7.4 for 30 min is 2.0 to 5.0 times the amount that binds to intact type I collagen membranes after incubation for 30 min. S5: The dual-response linker layer contains both matrix metalloproteinase-sensitive linker units and reactive oxygen species-sensitive linker units. The matrix metalloproteinase-sensitive linker units contain peptides that can be cleaved by at least one of MMP-1, MMP-3, and MMP-9. The reactive oxygen species-sensitive linker units contain thioacetyl linker groups. S6: The partitioned release kernel is set as a peripheral release region, an intermediate release region and a kernel release region. The peripheral release region is loaded with oxidative glycation buffer components, the intermediate release region is loaded with network degradation buffer components, and the kernel release region is loaded with collagen arrangement support components. S7: The collagen affinity response layer containing dual-gated vesicles of collagen damage microenvironment, together with the stratum corneum hydration flux regulation layer and the surface water-retaining film layer, are used to construct a non-invasive anti-aging skin care system. The stratum corneum hydration flux regulation layer, collagen affinity response layer and surface water-retaining film layer are arranged in sequence from the skin contact side to the outside.

2. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 1, characterized in that, In step S1, the skin layering parameters are obtained as follows: dermal echo density is acquired using 20-50MHz high-frequency ultrasound, skin rebound rate is acquired using a skin elasticity tester, transdermal water loss is acquired using a transdermal water loss tester, stratum corneum water content is acquired using a stratum corneum water content tester, and erythema index is acquired using a skin colorimeter. Before testing, the test skin was equilibrated for 20-30 minutes in an environment with a temperature of 22-25℃ and a relative humidity of 45%-55%. The skin layer state parameters were measured three times consecutively in the same test area and the arithmetic mean was taken.

3. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 2, characterized in that, In step S1, the dermal echo density and skin rebound rate of the test skin detection area are compared with the corresponding parameters of the non-exposed area on the inner forearm of the same test subject. Items with dermal echo density lower than 80% of the non-exposed area of ​​the inner forearm, skin rebound rate lower than 75% of the non-exposed area of ​​the inner forearm, transdermal water loss greater than 15 g / (mh), stratum corneum water content less than 35 a.u., and erythema index greater than 20 are classified as abnormal items. When there are 0 or 1 abnormal items, it is determined to be a Level 1 collagen network risk level; when there are 2 or 3 abnormal items, it is determined to be a Level 2 collagen network risk level; when there are 4 or 5 abnormal items, it is determined to be a Level 3 collagen network risk level. In step S2, when the collagen network risk level is level one, the damaged collagen recognition peptide accounts for 0.5% to 1.5% of the total lipid molar amount in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker unit to reactive oxygen species-sensitive linker unit is 1:1 to 1.5:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 5 to 7:2 to 3:1 to 2. When the collagen network risk level is level 2, the collagen recognition peptide accounts for 1.5% to 3.0% of the total lipid molars in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units is 1.5:1 to 2.5:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 3 to 5:3 to 4:2 to 3. When the collagen network risk level is level three, the damaged collagen recognition peptide accounts for 3.0% to 5.0% of the total lipid molars in the lipid biomimetic membrane layer, the molar ratio of matrix metalloproteinase-sensitive linker units to reactive oxygen species-sensitive linker units is 2.5:1 to 4:1, and the mass ratio of oxidative glycation buffer component, network degradation buffer component, and collagen arrangement support component is 2 to 3:4 to 5:3 to 4.

4. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 3, characterized in that, In step S3, the collagen damage microenvironment dual-gated vesicles are prepared as follows: the lipid biomimetic membrane material is dissolved in ethanol, and the ethanol is removed under reduced pressure to form a lipid film; An aqueous phase containing an oxidative glycation buffer component, a network degradation buffer component, and a collagen arrangement support component was added to a lipid membrane for hydration to obtain a primary emulsion. The primary emulsion was homogenized by microfluidic jetting 3 to 6 times at 60 to 100 MPa, and then granulated using a polycarbonate membrane with a pore size of 100 to 200 nm to obtain basic vesicles. A damaged collagen recognition peptide with a cysteine ​​linker, a matrix metalloproteinase-sensitive linker with a maleimide terminal, and a reactive oxygen species-sensitive linker with a thioketal linker are added to the basic vesicles. This allows the damaged collagen recognition peptide to link to the outer side of the lipid biomimetic membrane, and the matrix metalloproteinase-sensitive linker and the reactive oxygen species-sensitive linker to link between the lipid biomimetic membrane and the partitioned release core.

5. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 4, characterized in that, The raw materials for the lipid biomimetic membrane layer include, by mass, 45-65 parts hydrogenated lecithin, 10-25 parts cholesterol, 1-8 parts ceramide NP, 0.2-3 parts phytosphoin, 0.5-5 parts sodium stearoyl glutamate, and 2-10 parts squalane; The D50 size of the dual-gated vesicles in the collagen damage microenvironment is 50–120 nm, the polydispersity index is no greater than 0.25, and the zeta potential is -5 mV to -30 mV.

6. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 5, characterized in that, In step S4, the damaged collagen recognition peptide is a single-chain collagen mimic peptide, which contains 6 to 10 Gly-Pro-Hyp repeating units arranged in succession. The ends of the single-chain collagen mimic peptide are connected to cysteine ​​linkage groups, which are connected to the lipid biomimetic membrane layer.

7. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 6, characterized in that, In step S5, the matrix metalloproteinase-sensitive linker includes one of the following: Gly-Pro-Leu-Gly-Val-Arg-Gly peptide, Pro-Leu-Gly-Leu-Ala-Gly peptide, and Gly-Pro-Gln-Gly-Ile-Trp-Gly-Gln peptide. One end of the matrix metalloproteinase-sensitive linker is connected to the lipid biomimetic membrane layer, and the other end is connected to the partitioned release core. The reactive oxygen species-sensitive linker includes a thioacetate linker, which is disposed in the outer coating material of the partitioned release core.

8. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 7, characterized in that, In step S6, the oxidative glycation buffer component is composed of ergothioneine, carnosine and ferulic acid glucoside, and the mass ratio of ergothioneine, carnosine and ferulic acid glucoside is 1-3:2-5:1-3. The network degradation buffer component is composed of epigallocatechin gallate, oligomeric proanthocyanidins and low molecular weight sodium hyaluronate, and the mass ratio of epigallocatechin gallate, oligomeric proanthocyanidins and low molecular weight sodium hyaluronate is 3-6:2-4:1-3. The collagen arrangement support component consists of tripeptide-10 citrulline, acetyl tetrapeptide-9, and palmitoyl tripeptide-5, with a mass ratio of 2–4:2–4:1–3.

9. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 8, characterized in that, In step S7, the stratum corneum hydration flux regulating layer comprises, by mass percentage, 0.5%–3.0% betaine, 0.1%–1.0% ectoine, 0.5%–3.0% trehalose, 0.05%–0.5% lactobionic acid, and the balance water, with a pH of 5.2–5.8 and a viscosity of 500–3000 mPas; The collagen affinity response layer comprises, by mass percentage, 0.05%–2.0% dual-gated vesicles of collagen damage microenvironment, 2.0%–8.0% glycerol, 2.0%–8.0% 1,3-propanediol, and the balance water, with a pH of 5.5–6.2 and a viscosity of 800–5000 mPas; The surface water-retaining film layer comprises, by mass percentage, 0.05%–0.3% low molecular weight sodium hyaluronate, 0.05%–0.5% β-glucan, 0.2%–2.0% pullulan, and the balance water. The dry film thickness after film formation is 2–15 μm, and the film formation time is 3–10 min. The non-invasive anti-aging skincare system is free of azone, dimethyl sulfoxide, and ethanol (greater than 5% by weight).

10. The method for constructing a non-invasive anti-aging skincare system that targets and protects the dermal collagen network according to claim 9, characterized in that, The cumulative release rate of dual-gated vesicles in the collagen damage microenvironment was no more than 25% after 6 hours in phosphate buffer at pH 7.

4. The cumulative release rate in phosphate buffer containing 50–200 ng / mL matrix metalloproteinases was 35%–55% over 6 hours. The cumulative release rate was 55%–80% over 6 hours in phosphate buffer containing 50–200 ng / mL matrix metalloproteinases and 100–500 mol / L hydrogen peroxide. Matrix metalloproteinases are one of MMP-1, MMP-3, and MMP-9.

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