Detachable high-flux skin chip and construction method of inflammatory skin model thereof

Abstract

The invention discloses a detachable high-flux skin chip and a construction method of an inflammation skin model thereof. The detachable high-flux skin chip sequentially comprises a top sealing layer, an upper-layer chip, a porous membrane, a lower-layer chip and a bottom sealing layer from top to bottom, the upper-layer chip and the lower-layer chip are respectively provided with three groups of * 3-chamber high-throughput independent culture arrays, and each group of high-throughput independent culture array comprises three cell culture chambers which are connected in series and supports synchronous perfusion of culture media containing different concentrations of drugs or different types of drugs; and the upper and lower layers of high-flux independent culture arrays are respectively connected in series through the upper-layer tree-shaped channel and the lower-layer tree-shaped channel. The microfluidic technology and tissue engineering are combined, a 3 * 3 high-flux independent culture array is integrated through modular detachable design, single-time nine-model parallel screening is achieved, the bottleneck of the prior art is broken through cooperatively, the microenvironment is simulated dynamically, atopic dermatitis pathological characteristics are reproduced, and the authenticity, flux and flexibility of skin inflammation research are improved.

Description

Technical Field

[0001] This invention relates to the fields of microfluidics and tissue engineering, specifically to a detachable high-throughput skin chip and a method for constructing an inflamed skin model thereon. Background Technology

[0002] As the largest physiological barrier organ in the human body, the pathological mechanisms of inflammatory diseases of the skin and the development of new drugs have long relied on traditional animal models. However, animal models have problems such as differences in species immune responses, heterogeneity in skin structure, and differences in drug metabolism pathways, resulting in low efficiency in the clinical translation of research results.

[0003] Atopic dermatitis (AD) is one of the most common chronic inflammatory skin diseases, with a heavy global disease burden. However, its pathogenesis has not been fully elucidated, and the development of therapeutic drugs is limited by the lack of highly biomimetic models.

[0004] While existing skin chips have made improvements in nutrient supply, they still fall short in simulating the dynamic inflammatory microenvironment required for atopic dermatitis and supporting high-throughput drug screening. Traditional Transwell chamber or single-chamber chips can usually only perform single-point experiments, making it difficult to test multiple combinations of inflammatory factors or drug concentration gradients simultaneously. Furthermore, the number of samples obtained in a single experiment is limited, resulting in a relatively long drug screening cycle and high costs.

[0005] Therefore, developing a skin chip that can simulate the dynamic physiological microenvironment of the skin and support high-throughput parallel detection has become a key technological requirement to overcome the limitations of static culture systems and improve the efficiency of research on skin inflammation mechanisms such as Alzheimer's disease and drug screening. Summary of the Invention

[0006] The purpose of this invention is to address the problems of insufficient biomimicry of skin models, lack of dynamic inflammatory microenvironment simulation, low experimental throughput and poor repeatability, and insufficient application flexibility in the prior art. This invention proposes a detachable high-throughput skin chip and a method for constructing an inflammatory skin model thereon.

[0007] To achieve the above objectives, the technical solution specifically adopted by the present invention is as follows: A detachable high-throughput skin chip comprises, from top to bottom, a top sealing layer, an upper chip, a porous membrane, a lower chip, and a bottom sealing layer; wherein: The upper and lower chips are fabricated using PDMS molding through 3D printing molds. Three sets of three-chamber high-throughput independent culture arrays are designed. Each set of high-throughput independent culture arrays contains three cell culture chambers connected in series. The three sets of high-throughput independent culture arrays are independent of each other and support the simultaneous perfusion of culture medium containing different concentrations or different types of drugs. The upper and lower high-throughput independent culture arrays are connected in series through upper and lower dendritic channels, respectively. The porous membrane is one of PS membrane, PC membrane, PET membrane, and PVDF membrane, and is located between the upper chip and the lower chip, separating the cell culture chambers of the upper chip and the lower chip, thereby forming an upper culture chamber and a lower culture chamber; The top sealing layer is provided with an upper channel inlet, an upper channel outlet, and an upper chip fixing hole corresponding to the upper chip; the bottom sealing layer is provided with a lower channel inlet, a lower channel outlet, and a lower chip fixing hole corresponding to the lower chip.

[0008] In the detachable high-throughput skin chip of this invention, both the upper and lower dendritic channels are three-level dendritic channels. The ends of each three-level dendritic channel consist of four branch channels, and the gas flow direction and velocity of all channels are parallel. The upper and lower dendritic channels are arranged in a 90° perpendicular cross pattern to simulate the skin tissue-blood vessel interaction. By independently controlling the perfusion parameters of the lower channels in the three arrays, test environments for the same drug at the same concentration, different concentration gradients of the same drug, and different concentrations of three different drugs can be simultaneously constructed.

[0009] In the detachable high-throughput skin chip of this invention, the channel depths of both the upper and lower tree-like channels are less than the thicknesses of the upper and lower chips. The top and bottom sealing layers are both made of transparent material, selected from PMMA, PDMS, COC / COP, PC, and PS.

[0010] The detachable high-throughput skin chip of the present invention can be used to construct an inflamed skin model, comprising the following steps: S1. Add the biological scaffold material containing fibroblasts to the cell culture chamber, and generate a dermal model after the biological scaffold material solidifies. S2. Keratinocytes are seeded onto the surface of the dermal model, and epidermal cells are differentiated and matured through gas-liquid culture to obtain a full-thickness skin model. S3. Introduce inflammatory factors into the full-thickness skin model to induce the construction of an in vitro model of atopic dermatitis; S4. Introduce the drug into the aforementioned in vitro model of atopic dermatitis to verify the anti-inflammatory effect of the drug.

[0011] Furthermore, the bioscaffold material is selected from at least one of collagen matrix, Matrigel matrix gel, gelatin, GelMA hydrogel, sodium alginate, and hyaluronic acid.

[0012] Further, in step S2, the gas-liquid culture involves perfusing sterile air into the upper chip dendritic channel and perfusing epidermal differentiation culture medium into the lower chip dendritic channel. The epidermal differentiation culture medium is selected from at least one of DMEM culture medium, DMEM / F12 culture medium, insulin, transferrin, CaCl2 solution, adenine, EGF, fetal bovine serum, vitamin C, and hydrocortisone.

[0013] Furthermore, the inflammatory factor is selected from at least one of TNF-α, IL-4, IL-13, IL-25, and LPS.

[0014] Furthermore, the drug is selected from at least one of curcumin, dexamethasone, hydrocortisone, and dupilumab.

[0015] The detachable high-throughput skin chip described in this invention can also be applied to in vitro research on the pathological mechanism of atopic dermatitis, drug testing, drug screening, and toxicity assessment.

[0016] This invention has the following characteristics and beneficial effects:

[0017] 1) Adopting a detachable five-layer modular architecture reduces experimental costs. The chip adopts a modular design, consisting of a top sealing layer, an upper chip, a porous membrane, a lower chip, and a bottom sealing layer. Each layer is secured with screws and mounting holes, allowing for repeated disassembly and sterilization. Simultaneously, the transparent material enables real-time observation of cell morphology using an inverted microscope, simplifying experimental monitoring procedures. Low-cost materials and standardized preparation processes further reduce research and development and application costs.

[0018] 2) Employing a 3×3 high-throughput culture array improves experimental efficiency. Both the upper and lower chips are designed with three independent culture units, each containing three interconnected culture chambers. Within each unit, the units are connected via recessed three-tiered tree-like channels, while maintaining physical isolation between units. This chip structure supports multiple parallel experiments (such as different drug concentrations or combinations of inflammatory factors), and can simultaneously construct nine skin models in a single experiment. Furthermore, the standardized microchannel design ensures consistency within each unit, improving the efficiency and reliability of drug screening and evaluation.

[0019] 3) Dynamically simulate skin structure to enhance biomimetic performance. This invention uses a multi-layer chip structure to simulate the epidermis, dermis, and microchannel vascular network. The tree-like microchannel network ensures uniform fluid distribution, and combined with gas-liquid interface culture, it enables the expression of epidermal differentiation markers (keratin 14 and filaggrin), thus realizing the biomimetic construction of skin tissue structure and function and effectively improving the physiological relevance of the model.

[0020] 4) Targeted construction of an in vitro model of atopic dermatitis In the gas-liquid interface culture stage, this invention induces abnormal differentiation and inflammatory response of keratinocytes within the chip by perfusing culture medium containing inflammatory factors through the lower dendritic channel, successfully reproducing the key pathological features of atopic dermatitis. At the same time, it supports real-time monitoring of inflammatory factor secretion, providing a reliable platform for AD mechanism research and drug evaluation.

[0021] In summary, this invention combines microfluidic technology with tissue engineering, integrating a 3×3 high-throughput independent culture array through a modular and detachable design to achieve parallel screening of 9 models in a single run. This collaboratively overcomes existing technological bottlenecks, dynamically reproducing AD pathological features in a biomimetic microenvironment. It supports testing of multiple drugs, drug concentrations, and inflammatory factors; and provides a complete workflow from inflammation induction to efficacy verification, significantly improving the authenticity, throughput, and flexibility of skin inflammation research. It provides an effective platform for the study of atopic dermatitis mechanisms and high-throughput initial screening of anti-inflammatory drugs. Attached Figure Description

[0022] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings: Figure 1 This is an exploded view of a detachable high-throughput skin chip according to an embodiment of the present invention; Figure 2 This is a top view schematic diagram of a detachable high-throughput skin chip according to an embodiment of the present invention; Figure 3 This is a three-dimensional schematic diagram of a detachable high-throughput skin chip according to an embodiment of the present invention; Figure 4 This is a flowchart illustrating the construction process of an in vitro skin model based on a microfluidic chip, according to an embodiment of the present invention. Figure 5 This is an H&E staining morphology diagram of an in vitro skin model based on a microfluidic chip, according to an embodiment of the present invention. Figure 6 This is an image showing the immunofluorescence staining results of specific marker expression in an in vitro skin model based on a microfluidic chip, according to an embodiment of the present invention. Figure 7 This image shows the qPCR analysis results of related gene expression in a model of atopic dermatitis and drug treatment constructed using a detachable high-throughput skin chip according to an embodiment of the present invention. Detailed Implementation

[0023] The present invention will now be described in detail with reference to specific embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0024] Example 1 A method for fabricating a detachable high-throughput skin chip includes the following steps: (1) Chip structure design: such as Figure 1 and Figure 2 As shown, the chip consists of five layers from top to bottom: a top sealing layer 1, an upper chip 2, a porous film 3, a lower chip 4, and a bottom sealing layer 5. The porous membrane 3 is disposed between the upper chip 2 and the lower chip 4, separating the upper culture chamber 22 of the upper chip 2 and the lower culture chamber 42 of the lower chip 4 to achieve physical separation of different culture chambers; the porous membrane 3 also serves as a common substrate for the dermis and epidermis, and the micropores evenly distributed on the membrane allow nutrients, oxygen and metabolic waste to diffuse in both directions, while preventing cell migration and maintaining the stability of the layered structure.

[0025] To achieve high throughput for drug testing, the upper culture chamber 22 and the lower culture chamber 42 are designed as three sets of three independent high-throughput culture arrays. Each array contains three cell culture chambers connected in series. The three chambers of each cell culture chamber are connected in series through a tree-like channel to achieve sharing of the microenvironment within the group. The three groups are completely independent of each other and support parallel control experiments.

[0026] To ensure sufficient nutrient delivery to cells during culture and to provide a near-in vivo microenvironment for cell proliferation, differentiation, and functional expression, the upper chip 2 and lower chip 4 are connected in series via upper and lower dendritic channels, respectively. Both the upper and lower dendritic channels employ a tertiary dendritic structure, with channel widths ranging from 200-2000 μm and heights from 200-1000 μm. In this embodiment, the primary channel has a width of 1000 μm, the secondary channel has a width of 800 μm, and the tertiary channel has a width of 600 μm and a height of 800 μm. The tertiary dendritic channel terminates in four branch channels, with all channels exhibiting parallel gas flow and equal velocity to simulate the skin tissue-blood vessel interaction.

[0027] To facilitate real-time observation of the cultivation process, both the top sealing layer 1 and the bottom sealing layer 5 are made of transparent material, selected from PMMA, PDMS, COC / COP, PC, and PS.

[0028] In order to achieve continuous perfusion of culture medium and sterile air during the cultivation process, the top sealing layer 1 is designed with an upper channel inlet 11, an upper channel outlet 13 and an upper chip fixing hole 12, and the bottom sealing layer 5 is designed with a lower channel inlet 51, a lower channel outlet 53 and a lower chip fixing hole 52.

[0029] (2) Chip mold preparation: The chip mold is prepared by 3D photopolymerization printing. Specifically, the 3D printing slice parameters are set as follows: layer thickness range is 0.1-0.5 mm, normal exposure time range is 2-10 s, light-off time range is 0.1-1.0 s, Z-axis lifting height range is 1-5 mm, and Z-axis lifting and retraction speed range is 10-30 mm / s. In this embodiment, the layer thickness is 0.1 mm, the normal exposure time is 2 s, the light-off time is 0.5 s, the Z-axis lifting height is 3 mm, and the Z-axis lifting and retraction speed is 20 mm / s.

[0030] Photosensitive resin is selectively cured using ultraviolet light, and the cured resin is deposited layer by layer until a complete 3D mold is formed. The printed mold is then peeled off from the printing substrate, excess resin is washed off with industrial alcohol, and it is left to air dry at room temperature. Finally, the mold is cured under an ultraviolet lamp for 5-20 minutes to obtain a complete chip mold.

[0031] (3) Preparation of upper and lower layer chips: The upper and lower layer chips are prepared by PDMS molding using a mold. Specifically, PDMS prepolymer and curing agent are mixed at a ratio of 10:1 to 20:1, degassed under vacuum, and then poured into the mold. The mixture is cured at 60-80℃ for 0.5-2 h. Finally, the chips are peeled off from the mold and cut to the appropriate size to obtain upper layer chip 2 and lower layer chip 4. In this embodiment, the ratio of PDMS prepolymer to curing agent is 10:1. After vacuum degassing, the mixture is cured at 80℃ for 2 h.

[0032] (4) Porous membrane bonding: The prepared upper chip 2 and porous membrane 3 are treated with a plasma cleaner (100-500 mA, 200-800 V) for 1-5 min. After cleaning, the porous membrane is quickly attached to the upper chip 2 to cover all the upper culture chambers 22. Then the lower chip 4 is treated under the same conditions for 1-5 min. Finally, the upper chip 2 and the lower chip 4 are manually aligned and the surface dust is cleaned with plastic tape for later use. The upper dendritic channel 21 and the lower dendritic channel 41 of the upper chip 2 and the lower chip 4 are arranged in a 90° perpendicular cross to simulate the skin tissue-blood vessel interaction.

[0033] (5) Assembly of microfluidic chip: Fix the dispensing needle to 12 locations on the top sealing layer 1 and bottom sealing layer 5, namely the upper channel inlet 11, the lower channel inlet 51, the upper channel outlet 13, and the lower channel outlet 53. Then assemble the chip in the order of top sealing layer 1, upper chip 2, porous membrane 3, lower chip 4, and bottom sealing layer 5. Fix it with screws. The screws pass through the upper chip fixing hole 12 and the lower chip fixing hole 52 on the top sealing layer 1 and bottom sealing layer 5, for a total of 16 locations. Connect the upper channel inlet 11, the lower channel inlet 51, the upper channel outlet 13, and the lower channel outlet 53 to the silicone tubes respectively.

[0034] The culture system mainly consists of an injection pump, a chip, a three-way valve, and a storage bottle, which are connected in sequence by silicone tubing. The upper channel inlet 11, the lower channel inlet 51, the upper channel outlet 13, and the lower channel outlet 53 are each connected to a three-way valve, which not only facilitates flexible switching between different fluids but also greatly simplifies the cell seeding process.

[0035] To prevent air bubbles from clogging the microchannels during the experiment, the culture medium was perfused using an extraction method. Six silicone tubing tubes, including the upper channel inlet 11 and the lower channel inlet 51, were inserted into the bottom of the centrifuge tube containing the culture medium. Six silicone tubing tubes, including the upper channel outlet 13 and the lower channel outlet 53, were connected to the injection pump to prevent air bubbles in the culture medium from being drawn into the culture system.

[0036] Example 2 A method for constructing an in vitro skin model based on a microfluidic chip includes the following steps: (1) Sterilization of microfluidic chips: Sterilization of microfluidic chips is an important step to ensure experimental accuracy and a sterile environment. Ultraviolet (UV) sterilization was used on the top sealing layer 1, upper chip 2, porous membrane 3, lower chip 4, and bottom sealing layer 5. Surface impurities were removed with tape, and the chips were disinfected by spraying with 75% alcohol and then placed in a clean bench for UV irradiation for at least 2 hours. The chips were then arranged... Figure 3 After assembly, 75% ethanol was perfused into the upper culture chamber 22 and the lower culture chamber 42 through the upper dendritic channel 21 and the lower dendritic channel 41, respectively, for 30-60 min. Then, the perfusion was replaced with PBS buffer solution for another 30-60 min, and finally, the perfusion was replaced with culture medium for another 30-60 min. In this example, the perfusion time was 30 min for all samples.

[0037] (2) Construction of the dermis: such as Figure 4As shown, to simulate the 3D growth environment of fibroblasts in vivo, human primary fibroblasts were embedded in a biological scaffold material for culture. First, the biological scaffold was diluted to a working concentration, and then mixed with at least one of DMEM basal medium, CaCl2 solution, and 10× PBS buffer to prepare the dermal matrix.

[0038] After mixing human primary fibroblasts with dermal matrix, 100-400 μL can be added to the upper chamber 22 of the chip culture and incubated in a cell culture incubator for 30-90 min. In this example, 200 μL is added to the upper chamber 22 of the chip culture and incubated in a cell culture incubator for 60 min.

[0039] After the dermal matrix solidifies, DMEM complete culture medium is poured into both the upper dendritic channel 21 and the lower dendritic channel 41 at a flow rate of 1-10 μL / min. In this example, the flow rate is 1 μL / min.

[0040] (3) Construction of the epidermal layer: Primary human keratinocytes were seeded into the upper dermal layer of the culture chamber 22 through a three-way valve and left to stand for 4-6 h. After the cells adhered to the surface of the dermal matrix, epidermal cell culture medium was perfused into the upper dendritic channel 21 and DMEM complete culture medium was perfused into the lower dendritic channel 41. The flow rate was 1-10 μL / min. In this example, the flow rate was 1 μL / min.

[0041] (4) Gas-liquid interface culture: After the keratinocytes have grown to complete fusion, remove the culture medium in the upper channel 22, introduce sterile air, and replace the medium in the lower dendritic channel 41 with epidermal differentiation culture medium. Culture the cells under gas-liquid interface conditions to induce cell differentiation.

[0042] (5) Model validation i. Histological analysis: The constructed skin model was removed from culture chamber 22, fixed in 4% paraformaldehyde for 20 min, dehydrated with graded alcohol, embedded in paraffin, sectioned (5 μm thick), and stained with hematoxylin and eosin (HE). Results are as follows: Figure 5 The constructed skin model successfully formed skin tissue with a distinct layered structure, including the epidermis and dermis. The epidermis clearly shows the basic layered morphology of the stratum corneum, stratum granulosum, stratum spinosum, and stratum basale. The stratum basale exhibits a regular arrangement of a single layer of columnar epithelial cells, which eventually differentiate into the stratum corneum as they migrate towards the surface. In the dermis, fibroblasts are evenly distributed within the biological scaffold material, exhibiting a typical spindle-shaped morphology, with extracellular matrix filling the intercellular spaces.

[0043] ii. Immunofluorescence analysis: After antigen retrieval, skin model sections were stained with primary antibodies against keratin 14 and filaggrin, and their corresponding secondary antibodies, respectively. The immunofluorescence staining results are shown below. Figure 6 The results showed that keratin 14 was continuously and strongly expressed in the basal layer, proving that the basal cells in the model maintained an undifferentiated state, consistent with the physiological characteristics of normal epidermis. The stratum corneum-specific marker filaggrin showed continuous lamellar enrichment, indicating that the keratinocytes in the model had completed the terminal differentiation process and the stratum corneum structure was mature.

[0044] Example 3 A method for constructing an Alzheimer's disease (AD) inflammation model and validating drugs based on a microfluidic chip includes the following steps: (1) Induction of inflammation model: In order to construct a skin inflammation model and simulate the pathological features of AD, during the last three days of gas-liquid culture, inflammatory factors (10-50 ng / mL, in this example, the amount of inflammatory factors used was 20 ng / mL) were added to the epidermal differentiation medium and perfused into the lower culture chamber 42 through the lower channel 41 to promote the release of inflammatory mediators, thereby inducing the destruction of skin barrier function and inflammatory response.

[0045] (2) qPCR detection of inflammatory factors and barrier gene expression: Skin models were collected from culture chamber 22, and total RNA was extracted using TRIzol reagent, reverse transcribed into cDNA, and then subjected to qPCR. The expression levels of carbonic anhydrase 2 (CA2), filaggrin (FLG), and interleukin 6 (IL6) were analyzed using beta-actin (β-actin) as an internal control. Results are shown below. Figure 7 As shown, the expression levels of CA2 and IL-6 in the inflammatory factor treatment group were upregulated by 10.1-fold and 3.2-fold, respectively, compared with the control group, while the expression level of FLG was reduced by 16.7%, indicating that the inflammatory model successfully simulated the gene expression characteristics of keratinocytes in the inflammatory microenvironment.

[0046] (3) Verification of the anti-inflammatory effect of the drug: During the last three days of gas-liquid culture, differentiation medium containing inflammatory factors (10-50 ng / mL, in this example, the amount of inflammatory factors was 20 ng / mL) and the drug was perfused into the lower culture chamber 42 through the lower channel 41. The skin model in the upper culture chamber 22 was collected, and total RNA was extracted with TRIzol reagent, reverse transcribed into cDNA, and then qPCR was performed. Compared with the AD model group, the drug intervention group reduced IL-6 expression and partially reversed the abnormal expression of epidermal differentiation markers, suggesting that the drug can not only alleviate the inflammatory response, but may also maintain the skin barrier function by regulating epidermal differentiation-related genes. This model successfully simulated the induction and treatment of AD skin tissue inflammation and can be further applied to the screening of irritant compounds or the evaluation of drug treatment effects.

[0047] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely preferred examples and are not intended to limit the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. A detachable high-throughput skin chip, characterized in that, From top to bottom, it includes a top sealing layer, an upper chip, a porous membrane, a lower chip, and a bottom sealing layer; The upper and lower chips are designed with 3 sets of 3×3 high-throughput independent culture arrays. Each set of high-throughput independent culture arrays contains 3 cell culture chambers connected in series. The 3 sets of high-throughput independent culture arrays are independent of each other and support the synchronous perfusion of culture medium containing different concentrations or different types of drugs. The upper and lower high-throughput independent culture arrays are connected in series through the upper dendritic channel and the lower dendritic channel, respectively. The porous membrane is located between the upper chip and the lower chip, separating the cell culture chambers of the upper chip and the lower chip, thereby forming an upper culture chamber and a lower culture chamber; The top sealing layer is provided with an upper channel inlet, an upper channel outlet, and an upper chip fixing hole corresponding to the upper chip; the bottom sealing layer is provided with a lower channel inlet, a lower channel outlet, and a lower chip fixing hole corresponding to the lower chip.

2. The detachable high-throughput skin chip as described in claim 1, characterized in that, Both the upper and lower tree-like channels are three-level tree-like channels. The end of each three-level tree-like channel consists of four branch channels, and the gas flow direction of all channels is parallel and the flow velocity is equal. The upper and lower tree-like channels are arranged in a 90° perpendicular cross pattern to simulate the interaction between skin tissue and blood vessels.

3. The detachable high-throughput skin chip as described in claim 1, characterized in that, The channel depth of both the upper and lower tree-like channels is less than the thickness of the upper and lower chips.

4. The detachable high-throughput skin chip as described in claim 1, characterized in that, The porous membrane is one of PS membrane, PC membrane, PET membrane, and PVDF membrane.

5. The detachable high-throughput skin chip as described in claim 1, characterized in that, Both the top and bottom sealing layers are made of transparent material, selected from PMMA, PDMS, COC / COP, PC, and PS.

6. The application of the detachable high-throughput skin chip as described in any one of claims 1-5, characterized in that, The construction of an inflammatory skin model includes the following steps: S1. Add the biological scaffold material containing fibroblasts to the cell culture chamber, and generate a dermal model after the biological scaffold material solidifies. S2. Keratinocytes are seeded onto the surface of the dermal model, and epidermal cells are differentiated and matured through gas-liquid culture to obtain a full-thickness skin model. S3. Introduce inflammatory factors into the full-thickness skin model to induce the construction of an in vitro model of atopic dermatitis; S4. Introduce the drug into the aforementioned in vitro model of atopic dermatitis to verify the anti-inflammatory effect of the drug.

7. The application of the detachable high-throughput skin chip as described in claim 6, characterized in that, The bioscaffold material is selected from at least one of collagen matrix, Matrigel matrix gel, gelatin, GelMA hydrogel, sodium alginate, and hyaluronic acid.

8. The application of the detachable high-throughput skin chip as described in claim 6, characterized in that, In step S2, the gas-liquid culture involves perfusing sterile air into the upper chip's dendritic channels and perfusing epidermal differentiation culture medium into the lower chip's dendritic channels. The components of the epidermal differentiation culture medium are selected from at least one of DMEM culture medium, DMEM / F12 culture medium, insulin, transferrin, CaCl2 solution, adenine, EGF, fetal bovine serum, vitamin C, and hydrocortisone.

9. The application of the detachable high-throughput skin chip as described in claim 6, characterized in that, The inflammatory factors are selected from at least one of TNF-α, IL-4, IL-13, IL-25, and LPS; the drug is selected from at least one of curcumin, dexamethasone, hydrocortisone, and dupilumab.

10. The application of the detachable high-throughput skin chip as described in any one of claims 1-5, characterized in that, Used for in vitro research on the pathological mechanisms of atopic dermatitis, drug testing, drug screening, and toxicity assessment.

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