Microfluidic blood vessel chip, system, preparation method and application for intracranial aneurysm

Abstract

The application relates to an intracranial aneurysm microfluidic blood vessel chip, system, preparation method and application, and belongs to the medical technical field.The intracranial aneurysm microfluidic blood vessel chip comprises a Y-shaped microfluidic channel, a negative pressure suction cavity and an elastic film, the negative pressure suction cavity is used for being connected with a negative pressure suction device to vacuumize the negative pressure suction cavity, the Y-shaped microfluidic channel meets at a crossing, the crossing is provided with an opening, the elastic film is arranged at the opening and separates the negative pressure suction cavity from the Y-shaped microfluidic channel, the elastic film is a treated polyisoprene film, and the treatment comprises plasma treatment; when the negative pressure suction cavity generates negative pressure, the elastic film can bulge towards the negative pressure suction cavity, and has different bulging degrees under different sizes of negative pressure. The application has good biocompatibility, can simulate the continuous change of a tumor, and has consistency with the intracranial aneurysm in the body in terms of hemodynamic characteristics.

Description

Technical Field

[0001] This application relates to the field of medical technology, and more specifically, to a microfluidic vascular chip, system, preparation method and application for intracranial aneurysms. Background Technology

[0002] Intracranial aneurysms (IAs) are localized aneurysmal protrusions of the arterial wall caused by abnormal dilation of intracranial arteries. Vascular factors influencing IAs include the composition of the vessel wall, the size and shape of the aneurysm, the presence of a cyst, the location of the aneurysm, and hemodynamics. IAs commonly form at the bifurcation and sharp bends of the Circle of Willis arteries. The bifurcation of the MCA and the anterior communicating artery are the most common sites of IA rupture, possibly related to changes in hemodynamic magnitude at these locations.

[0003] Preventing vascular infarction (IA) rupture remains a clinical challenge. Previous studies have primarily used experimental animals to construct in vivo models. However, the formation and rupture of vascular-regulated IAs typically involve multiple complex and dynamically interacting factors. Using an entire experimental organism, the mixture of factors often hinders the exploration of related mechanisms, and interspecies differences between humans and animals can also impede research progress. In vitro models, on the other hand, offer advantages over in vivo models because they allow for precise control of conditions and the direct culture of human cells for experiments, enabling more effective control.

[0004] Current in vitro models mainly include 3D printing or microfluidic chips. 3D printing can directly and conveniently simulate the geometry of real aneurysms, and its flow measurement and CFD numerical simulation are very consistent with the actual situation in vivo, which can be used to study complex hemodynamic phenomena occurring within aneurysms. However, the materials used in this method still have limitations in terms of cell biocompatibility, and their mechanical properties differ significantly from those of living blood vessels. Although microfluidic chips have solved the above problems, existing microfluidic chips are still limited to completely fixed tumor size, unable to realize continuous changes in tumor size, and cannot meet the needs of in-depth research on pathological mechanisms and drug screening. Summary of the Invention

[0005] This application provides a microfluidic vascular chip, system, preparation method and application for intracranial aneurysms, which has good biocompatibility and can simulate the continuous changes of tumors. Its hemodynamic characteristics are consistent with those of intracranial aneurysms in vivo.

[0006] This application is implemented as follows:

[0007] In a first aspect, this application provides a microfluidic vascular chip for intracranial aneurysms, comprising: a Y-shaped microfluidic channel, a negative pressure suction cavity, and an elastic membrane. The negative pressure suction cavity is used to connect to a negative pressure suction device for evacuating the negative pressure suction cavity. The Y-shaped microfluidic channels intersect at the intersection, and an opening is provided at the intersection. The elastic membrane is disposed at the opening and separates the negative pressure suction cavity from the Y-shaped microfluidic channels. The elastic membrane is a treated polyisoprene film, and the treatment includes plasma treatment.

[0008] When a negative pressure is generated in the negative pressure suction cavity, the elastic membrane can bulge out into the negative pressure suction cavity, and the degree of bulging varies under different levels of negative pressure.

[0009] In one possible implementation, the treatment includes sequential plasma treatment and fibronectin cell incubation treatment.

[0010] In one possible implementation, the Y-shaped microfluidic channel includes a main channel and two branch channels, the central axes of the two branch channels being symmetrical about the central axis of the main channel and forming an angle of 110°.

[0011] In one possible implementation, the main channel and the two branch channels are both cylindrical channels with a diameter of 2 mm and circular openings of 2 mm.

[0012] In one possible implementation, the intracranial aneurysm microfluidic vascular chip is generally disc-shaped, including a first module with a Y-shaped microfluidic channel and a second module with a negative pressure suction cavity, the first module and the second module being connected to form a disc shape.

[0013] Secondly, this application provides a method for fabricating a microfluidic vascular chip for intracranial aneurysms, comprising the following steps:

[0014] The first module with a Y-shaped microfluidic channel was prepared by using a mold.

[0015] An opening is made at the intersection of the Y-shaped microfluidic channels;

[0016] The elastic membrane is placed over the opening; the preparation steps of the elastic membrane include: cleaning and sterilizing the polyisoprene film, followed by plasma treatment.

[0017] A second module with a negative pressure suction cavity is prepared, wherein the negative pressure suction cavity has a first connection port communicating with the negative pressure suction cavity and a second connection port for connecting with a negative pressure suction device.

[0018] The first connection port of the second module is connected to the opening of the first module, thereby connecting the second module to the first module, and the elastic membrane covers the opening and the first connection port.

[0019] In one possible implementation, the plasma treatment is oxygen plasma treatment, and the treatment time is 1 to 3 minutes.

[0020] In one possible implementation, the polyisoprene film is subjected to plasma treatment, followed by cell culture incubation using fibronectin.

[0021] Thirdly, this application provides an application of an intracranial aneurysm microfluidic vascular chip prepared by the method of the first aspect or the method of the second aspect in the screening of intracranial aneurysm drugs.

[0022] Fourthly, this application provides an intracranial aneurysm microfluidic vascular chip system, including an intracranial aneurysm microfluidic vascular chip, a circulation pump, a reservoir, and a negative pressure suction device. The negative pressure suction device is connected to a negative pressure suction cavity for evacuating the cavity. The reservoir is connected to the main channel of a Y-shaped microfluidic channel via a first connecting tube, and the reservoir is connected to two branch channels of the Y-shaped microfluidic channel via a second connecting tube. The circulation pump is connected to the first connecting tube and / or the second connecting tube.

[0023] The embodiments of this application have at least the following beneficial effects:

[0024] The intracranial aneurysm microfluidic vascular chip of this application features a Y-shaped microfluidic channel that resembles the bifurcated blood vessels in the brain, closely mimicking the hemodynamic environment of intracranial blood vessels. A negative pressure suction cavity is connected to a negative pressure suction device for evacuating the cavity. When negative pressure is generated in the cavity, an elastic membrane bulges outwards, exhibiting different degrees of bulging under varying negative pressures, achieving controllable and continuous cystic bulging. This simulates the dynamic process of intracranial aneurysm volume growth and outward bulging. Results demonstrate that the hemodynamic characteristics of the embodiments described in this application are consistent with those of in vivo intracranial aneurysms.

[0025] The method for preparing the intracranial aneurysm microfluidic vascular chip of this application involves assembling a first module with a Y-shaped microfluidic channel, a second module with a negative pressure suction cavity, and an elastic membrane to form an intracranial aneurysm microfluidic vascular chip. The assembly method of this preparation method is simple.

[0026] The intracranial aneurysm microfluidic vascular chip of this application can simulate the dynamic process of intracranial aneurysm volume growth and outward bulging, and can be used for effective screening of intracranial aneurysm drugs. Attached Figure Description

[0027] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0028] Figure 1 This is a schematic diagram of the structure of the intracranial aneurysm microfluidic vascular chip according to an embodiment of this application;

[0029] Figure 2 This is a physical image of the intracranial aneurysm microfluidic vascular chip according to an embodiment of this application;

[0030] Figure 3 This is a schematic diagram of the intracranial aneurysm microfluidic vascular chip system according to an embodiment of this application;

[0031] Figure 4 The stress-strain curves of the three membrane materials in Experiment Example 1 are shown.

[0032] Figure 5 The figure shows the contact angle test results from Experiment Example 2;

[0033] Figure 6 This is a diagram showing the immunofluorescence staining results from Experiment Example 3;

[0034] Figure 7 This is a diagram showing the degree of bulging of the elastic membrane in Example 1 under different negative pressure intensities;

[0035] Figure 8 The results are shown in the figure of the changes in flow field and streamlines in Experiment Example 5;

[0036] Figure 9 The results of the smooth muscle cell skeleton test in the implementation method;

[0037] Figure 10 The results are immunofluorescence staining results of the smooth muscle cell phenotype in the implementation method.

[0038] Icons: 10-Intracranial aneurysm microfluidic vascular chip; 11-Y-shaped microfluidic channel; 111-Main channel; 112-Bifurcation channel; 12-Negative pressure suction cavity; 13-Elastic membrane; 20-Intracranial aneurysm microfluidic vascular chip system; 22-Reservoir bottle; 23-Circulation pump; 24-Negative pressure suction device. Detailed Implementation

[0039] The embodiments of this application will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of this application. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0040] The following provides a detailed description of the intracranial aneurysm microfluidic vascular chip 10, its system, fabrication method, and applications according to embodiments of this application:

[0041] In a first aspect, embodiments of this application provide a microfluidic vascular chip 10 for intracranial aneurysms, comprising: a Y-shaped microfluidic channel 11, a negative pressure suction cavity 12, and an elastic membrane 13. The negative pressure suction cavity 12 is connected to a negative pressure suction device 24 for evacuating the negative pressure suction cavity 12. The Y-shaped microfluidic channels 11 intersect at an intersection with an opening. The elastic membrane 13 is disposed at the opening and separates the negative pressure suction cavity 12 from the Y-shaped microfluidic channels 11 (see reference). Figure 1 and Figure 2 When the negative pressure is generated in the negative pressure suction cavity 12, the elastic membrane 13 can bulge out into the negative pressure suction cavity 12, and the degree of bulging is different under different negative pressures.

[0042] Among them, the elastic membrane 13 is a polyisoprene film that has been treated, including plasma treatment.

[0043] The inventors of this application discovered in their research that the ease with which an elastic membrane material can be stretched determines its ability to form cystic bulges during a negative pressure suction process. The inventors of this application measured the tensile properties of three membrane materials: polyisoprene film, silicone rubber modified atmosphere film, and polydimethylsiloxane film. Figure 4 The results showed that polydimethylsiloxane film had a low elongation at break, while polyisoprene film had a low Young's modulus and a high elongation at break. Furthermore, the film material itself possessed good optical transparency, which is beneficial for observation and detection using optical methods such as fluorescence microscopy. Therefore, polyisoprene film was preliminarily selected as an elastic film material.

[0044] However, the inventors of this application discovered that polyisoprene films are hydrophobic, which is detrimental to cell adhesion and the wetting of the film surface by hydrogel precursor solutions. Therefore, plasma treatment of the polyisoprene film surface was chosen to improve its hydrophilicity. Exemplarily, oxygen plasma treatment was used for a treatment time of 1–3 minutes. Experimental results show that plasma treatment of the polyisoprene film surface significantly improved its hydrophilicity.

[0045] Furthermore, the polyisoprene film was subjected to plasma treatment followed by fibronectin cell incubation. Experimental results showed that plasma treatment followed by fibronectin cell incubation further enhanced the hydrophilicity of the elastic membrane. Moreover, the polyisoprene film treated with only oxygen plasma showed a small number of cells adhering, but the number was limited, and immunofluorescence staining of the cytoskeleton revealed poor cell spreading. In contrast, the polyisoprene film treated with plasma followed by fibronectin cell incubation exhibited significantly improved cell adhesion, a marked increase in the number of adhering cells, and immunofluorescence staining of the cytoskeleton showed that the adhering cells were well-spread, demonstrating good biocompatibility.

[0046] The intracranial aneurysm microfluidic vascular chip 10 of this application features a Y-shaped microfluidic channel 11 that resembles a bifurcated intracranial blood vessel, nearly physiologically mimicking the hemodynamic environment of intracranial blood vessels. A negative pressure suction cavity 12 is connected to a negative pressure suction device 24 for evacuating the cavity. When negative pressure is generated in the cavity 12, an elastic membrane 13 bulges outwards, exhibiting different degrees of bulging under varying negative pressures, achieving controllable and continuous cystic bulging and simulating the dynamic process of intracranial aneurysm volume growth and outward bulging. Results demonstrate that the hemodynamic characteristics of this embodiment are consistent with those of in vivo intracranial aneurysms.

[0047] In one embodiment, the intracranial aneurysm microfluidic vascular chip 10 is generally disc-shaped, comprising a first module with a Y-shaped microfluidic channel 11 and a second module with a negative pressure suction cavity 12. The first and second modules are connected to form a disc shape. The overall structure of the intracranial aneurysm microfluidic vascular chip 10 is stable, and the first and second modules are not easily separated or detached during experiments.

[0048] The Y-shaped microfluidic channel 11 includes a main channel 111 and two branch channels 112. The central axes of the two branch channels 112 are symmetrical about the central axis of the main channel 111 and form an angle of 110°. Optionally, the main channel 111 and the two branch channels 112 are both cylindrical channels with a diameter of 2 mm and a circular opening of 2 mm.

[0049] For example, the first module is made of polydimethylsiloxane, which is an organosilicon elastomer with stable chemical properties, good optical transparency and biocompatibility.

[0050] Secondly, this application provides a method for preparing an intracranial aneurysm microfluidic vascular chip 10 according to the first aspect, comprising the following steps:

[0051] (1) A first module with a Y-shaped microfluidic channel was prepared by means of a mold.

[0052] A mold for an intracranial aneurysm was fabricated using 3D printing to simulate the bifurcation of a cerebral artery. Specifically, a Y-shaped semi-cylindrical mold with a diameter of 2 mm and a bifurcation angle of 110° was designed.

[0053] Polydimethylsiloxane (PDMS) and curing agent are mixed in a ratio of 10:1 (w / w), degassed to remove air bubbles introduced during the mixing process, and then poured into a semi-cylindrical mold.

[0054] The mold was heated and cured at 60°C for 4 hours. After removing and cleaning the PDMS module, its surface was treated with oxygen plasma for 90 seconds. Two PDMS modules were then bonded together to form the first module with a Y-shaped microfluidic channel 11, followed by autoclaving. Plasma treatment significantly improved the surface adhesion of the PDMS modules. Furthermore, when accurately bonding two identical PDMS modules facing each other, baking at 60°C was used to solidify the bond. This process achieved leak-proof and seepage-proof microfluidic chip liquids while preserving the high transparency of PDMS.

[0055] (2) An opening is made at the intersection of the Y-shaped microfluidic channels. And each of the three top ends of the Y-shaped microfluidic channel has a connection hole.

[0056] (3) Cover the opening with the elastic membrane. It should be noted that the selection and treatment of the elastic membrane have been described in detail above, and therefore will not be repeated here.

[0057] (4) A second module with a negative pressure suction cavity is prepared, wherein the negative pressure suction cavity has a first connection port communicating with the negative pressure suction cavity and a second connection port for connecting with the negative pressure suction device.

[0058] For example, based on the disc-shaped configuration formed by the connection of the first and second modules, and considering the fan-shaped space reserved during the design of the first module, a 110° fan-shaped negative pressure suction cavity (transparent cavity) is designed at the bifurcation point. According to the design, a 1mm thick transparent acrylic sheet is cut using a laser engraving machine, and the joints of the assembled sheet are coated with PDMS to ensure airtightness. A 1mm radius circular first connection port is designed at the bottom of the negative pressure suction cavity, and a 3.5mm radius circular second connection port is designed at the top of the negative pressure suction cavity. A latex tube can be inserted into the second connection port to connect an external negative pressure suction device 24.

[0059] (5) Connect the first connection port of the second module to the opening of the first module, so that the second module is connected to the first module, and the elastic membrane covers the opening and the first connection port. After the second module is connected to the first module, the whole is in the shape of a disc, and the connection is covered with PDMS to ensure airtightness.

[0060] Alternatively, the maximum volume that the elastic membrane 13 can bulge in the intracranial aneurysm microfluidic vascular chip 10 can be defined as 100%, and the formula for calculating the spherical cap can be used: V=π / 3×(3×RH)·H 2 (V: volume of the spherical cap; R: radius of the sphere cut off from the spherical cap; H: height of the spherical cap), calculate the indentation height of the elastic membrane 13 when the bulging volume is 0%, 5%, 10%, 20%, 30%, 35%, 50%, and 100%, and set the negative pressure magnitude and pressure increase rate of the negative pressure suction device 24 so that the elastic membrane 13 can be stretched uniformly to the corresponding volume and kept stable:

[0061] 1) The experiment was divided into groups: control group: 0 bulging volume; 5% bulging group: negative pressure set at 5 kPa; 10% bulging group: negative pressure set at 10 kPa; 20% bulging group: negative pressure set at 20 kPa; 30% bulging group: negative pressure set at 30 kPa; 35% bulging group: negative pressure set at 35 kPa; 50% bulging group: negative pressure set at 45 kPa; 100% bulging group: negative pressure set at 55 kPa.

[0062] 2) The stretching speed for each group was 0.1 kPa / min, and the total duration of the experiment was 2 hours. The bulging elastic membrane 13 was gently removed for cytoskeleton determination and immunofluorescence staining for smooth muscle phenotype.

[0063] Among them, the experimental results corresponding to the smooth muscle cytoskeleton are as follows: Figure 9 As shown, the immunofluorescence staining results for the smooth muscle phenotype are as follows: Figure 10 As shown. From Figure 9 and Figure 10 The results showed that, with increasing stretching, smooth muscle cells tended to transform from a contractile phenotype to a synthetic phenotype (Str 10%), and from a synthetic phenotype to a pro-inflammatory senescent phenotype (Str 30%).

[0064] Thirdly, this application provides the application of an intracranial aneurysm microfluidic vascular chip prepared by the method of the first aspect or the method of the second aspect in the screening of intracranial aneurysm drugs.

[0065] The intracranial aneurysm microfluidic vascular chip of this application can simulate the dynamic process of intracranial aneurysm volume growth and outward bulging, and can be used for effective screening of intracranial aneurysm drugs.

[0066] Fourthly, this application provides an intracranial aneurysm microfluidic vascular chip system 20 (refer to...) Figure 3The first aspect includes an intracranial aneurysm microfluidic vascular chip 10, a circulation pump 23, a reservoir 22, and a negative pressure suction device 24. The negative pressure suction device 24 is connected to a negative pressure suction cavity 12 for evacuating the negative pressure suction cavity 12. The reservoir 22 is connected to the main channel 111 of the Y-shaped microfluidic channel 11 through a first connecting tube. The reservoir 22 is connected to the two branch channels 112 of the Y-shaped microfluidic channel 11 through a second connecting tube. The circulation pump 23 is connected to the first connecting tube and / or the second connecting tube.

[0067] The liquid in the storage bottle 22 is circulated through the Y-shaped microfluidic channel 11 by the circulation pump 23. The liquid first enters the main channel 111, then enters the two branch channels 112, and then returns to the storage bottle 22, thus forming a circulation. The negative pressure suction device 24 creates a vacuum in the negative pressure suction cavity 12, causing the elastic membrane 13 to bulge into the negative pressure suction cavity 12. The degree of bulging varies under different levels of negative pressure, achieving controllable and continuous cystic bulging. The hemodynamic characteristics are consistent with those of intracranial aneurysms in vivo.

[0068] The following detailed description of the intracranial aneurysm microfluidic vascular chip, system, preparation method and application of this application is provided in conjunction with the embodiments.

[0069] Example 1

[0070] This embodiment provides a microfluidic vascular chip 10 for intracranial aneurysms, which is generally disc-shaped. It consists of a first module with a Y-shaped microfluidic channel 11, an elastic membrane 13, and a second module with a negative pressure suction cavity 12. The Y-shaped microfluidic channel 11 includes a main channel 111 and two branch channels 112. The central axes of the two branch channels 112 are symmetrical about the central axis of the main channel 111, forming an angle of 110°. Both the main channel 111 and the two branch channels 112 are cylindrical channels with a diameter of 2 mm, and a circular opening with a diameter of 2 mm is provided at the intersection.

[0071] The elastic membrane 13 is located at the opening and separates the negative pressure suction cavity 12 from the Y-shaped microfluidic channel 11. When the negative pressure suction cavity 12 generates negative pressure, the elastic membrane 13 can bulge into the negative pressure suction cavity 12, and the degree of bulging is different under different negative pressures.

[0072] The fabrication steps of the above-mentioned intracranial aneurysm microfluidic vascular chip include:

[0073] (1) A first module with a Y-shaped microfluidic channel was prepared by means of a mold.

[0074] A Y-shaped semi-cylindrical mold with a diameter of 2 mm and a bifurcation angle of 110° was prepared using 3D printing.

[0075] Polydimethylsiloxane and curing agent are mixed in a ratio of 10:1 (w / w), degassed to remove air bubbles introduced during the mixing process, and then poured into a semi-cylindrical mold.

[0076] The mold is heated and cured at 60°C for 4 hours. After the PDMS module is removed and cleaned, the surface is treated with oxygen plasma for 90 seconds. The two PDMS modules are then bonded together and baked at 60°C to solidify the bond, forming the first module with Y-shaped microfluidic channels. Finally, the module is sterilized by high-pressure steam.

[0077] (2) An opening is made at the intersection of the Y-shaped microfluidic channels. And each of the three top ends of the Y-shaped microfluidic channel has a connection hole.

[0078] (3) Cover the opening with an elastic membrane. The preparation steps of the elastic membrane include: soaking the polyisoprene film in 75% alcohol for 10 min, and then irradiating it with ultraviolet light for 1 h for surface sterilization. Then, place it in a plasma cleaner and treat it with oxygen plasma for 90 s. Next, place it in a clean 3.5 cm culture dish and add 100 μg / mL of fibronectin (FN). Place the dish in a cell culture incubator and incubate at 37°C and 5% CO2 for 2 h.

[0079] (4) Based on the disc-shaped structure formed by connecting the first and second modules, and considering the fan-shaped space reserved during the design of the first module, a 110° fan-shaped negative pressure suction cavity (transparent cavity) is designed at the bifurcation point. According to the design, a 1mm thick transparent acrylic sheet is cut using a laser engraving machine, and the joints of the assembled sheet are coated with PDMS to ensure airtightness. A 1mm radius circular first connection port is designed at the bottom of the negative pressure suction cavity, and a 3.5mm radius circular second connection port is designed at the top of the negative pressure suction cavity, thus obtaining the second module.

[0080] (5) Connect the first connection port of the second module to the opening of the first module, so that the second module is connected to the first module, and the elastic membrane covers the opening and the connection port. After the second module is connected to the first module, the whole is in the shape of a disc, and the connection is covered with PDMS to ensure airtightness.

[0081] This embodiment also provides an intracranial aneurysm microfluidic vascular chip system 20, including the above-mentioned intracranial aneurysm microfluidic vascular chip 10, a circulation pump 23, a storage bottle 22, and a negative pressure suction device 24. The negative pressure suction device 24 is connected to the negative pressure suction cavity 12 through a second connecting hole for evacuating the negative pressure suction cavity 12. The storage bottle 22 is connected to the main channel 111 of the Y-shaped microfluidic channel 11 through a first connecting tube. The storage bottle 22 is connected to the two branch channels 112 of the Y-shaped microfluidic channel 11 through a second connecting tube. The circulation pump 23 is connected to the first connecting tube.

[0082] Example 2

[0083] This embodiment provides a microfluidic vascular chip for intracranial aneurysms, and its preparation method and system. The only difference from Embodiment 1 is that the elastic membrane in Embodiment 2 was not subjected to fibronectin cell culture incubation treatment.

[0084] Comparative Example 1

[0085] This comparative example provides a microfluidic vascular chip for intracranial aneurysms, as well as its preparation method and system. The only difference from Example 1 is that the elastic membrane of Comparative Example 1 was not subjected to plasma treatment and fibronectin cell culture incubation.

[0086] Experimental Example 1

[0087] Tensile properties of three film materials—polyisoprene film, silicone rubber modified atmosphere film, and polydimethylsiloxane film—were determined, and their stress-strain curves are shown below. Figure 4 As shown.

[0088] from Figure 4 As can be seen, polyisoprene film has a lower Young's modulus and a higher elongation at break, while polydimethylsiloxane film has a lower elongation at break.

[0089] Experimental Example 2

[0090] The elastic films prepared in Examples 1, 2, and 1 (Comparative Example 1) were placed at room temperature for 24 hours before contact angle testing was performed. The measurement results are as follows: Figure 5 As shown.

[0091] from Figure 5 It can be seen that the water contact angle of the untreated polyisoprene film is greater than 110°, indicating that the film material is hydrophobic; the water contact angle of the elastic film in Example 2 is 66°, indicating that plasma treatment can significantly improve the hydrophilicity of the polyisoprene film; the water contact angle of the elastic film in Example 1 is 37°, indicating that after oxygen plasma surface treatment and then incubation with fibronectin, the hydrophilicity of the polyisoprene film is further improved.

[0092] Experimental Example 3

[0093] The elastic membranes prepared in Examples 1, 2, and Comparative Example 1 were seeded with vascular endothelial cells. After 24 hours, the cytoskeleton (actin microfilaments, F-actin) was subjected to immunofluorescence staining. The results are as follows: Figure 6 As shown.

[0094] from Figure 6As can be seen, untreated polyisoprene films showed almost no cell adhesion; polyisoprene films treated with oxygen plasma alone showed a small number of cells attached, but the number was small, and immunofluorescence staining of the cytoskeleton showed poor spreading of the attached cells; in contrast, polyisoprene films treated with oxygen plasma and incubated with fibronectin showed significantly improved cell adhesion properties, with a significantly increased number of attached cells, and immunofluorescence staining of the cytoskeleton showed that the attached cells were in a better spreading state.

[0095] Test Example 4

[0096] Using the intracranial aneurysm microfluidic vascular chip system of Example 1, by controlling the negative pressure suction device to generate different negative pressure intensities, the elastic membrane exhibits different degrees of bulging, such as... Figure 7 As shown, this embodiment of the intracranial aneurysm microfluidic vascular chip can achieve controllable and continuous cystic bulging, simulating the dynamic process of IA volume growth and outward bulging.

[0097] Experimental Example 5

[0098] Using the intracranial aneurysm microfluidic vascular chip system of Example 1, different negative pressure intensities were generated by controlling the negative pressure suction device, resulting in different degrees of bulging of the elastic membrane. Based on the different set aneurysm bulging degrees, the changes in the flow field and streamlines within the aneurysm cavity were analyzed using ANSYS Fluent software. The results are as follows: Figure 8 As shown.

[0099] according to Figure 8 The results show that as the volume of the saccular aneurysm at the bifurcation increases, the wall shear stress (WSS) and intra-aneurysmal flow velocity significantly decrease, and the flow type changes from laminar flow under physiological conditions to turbulent flow. Based on CFD simulation analysis of clinical imaging data, it can be found that the hemodynamics of the intracranial aneurysm (IA) in vivo also undergoes several processes: laminar flow with a high WSS at the bifurcation of normal physiological cerebral blood vessels; in the early stages of IA formation, the IA is small in size, and some blood flows in to form turbulent flow, but the WSS remains at a high level; as the IA further expands, the blood flow velocity within the aneurysm decreases, forming turbulent flow with a low WSS. Therefore, it can be preliminarily demonstrated that the intracranial aneurysm microfluidic vascular chip of this embodiment, in simulating the occurrence and development of IA and the continuous increase in the bulging volume, exhibits changes in hemodynamic "mechanical intensity" from high WSS to low WSS and "flow type" from laminar flow to turbulent flow, which is consistent with the hemodynamic characteristics of the IA formation process in vivo.

[0100] The above are merely specific embodiments of this application and are not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A microfluidic vascular chip for intracranial aneurysms, characterized in that, It includes: The system comprises a Y-shaped microfluidic channel, a negative pressure suction cavity, and an elastic membrane. The negative pressure suction cavity is connected to a negative pressure suction device for evacuating the cavity. The Y-shaped microfluidic channels intersect at an opening. The elastic membrane is located at the opening and separates the negative pressure suction cavity from the Y-shaped microfluidic channels. The elastic membrane is a treated polyisoprene film, and the treatment includes plasma treatment. When the negative pressure suction cavity generates negative pressure, the elastic membrane can bulge into the negative pressure suction cavity, and the degree of bulging varies under different levels of negative pressure.

2. The intracranial aneurysm microfluidic vascular chip according to claim 1, characterized in that, The treatment includes sequential plasma treatment and fibronectin cell incubation.

3. The intracranial aneurysm microfluidic vascular chip according to claim 1 or 2, characterized in that, The Y-shaped microfluidic channel includes a main channel and two branch channels. The central axes of the two branch channels are symmetrical about the central axis of the main channel and form an angle of 110°.

4. The intracranial aneurysm microfluidic vascular chip according to claim 3, characterized in that, The main channel and the two branch channels are all cylindrical channels with a diameter of 2mm, and the openings are circular openings with a diameter of 2mm.

5. The intracranial aneurysm microfluidic vascular chip according to claim 1 or 2, characterized in that, The intracranial aneurysm microfluidic vascular chip is generally disc-shaped, including a first module with the Y-shaped microfluidic channel and a second module with the negative pressure suction cavity. The first module and the second module are connected to form a disc shape.

6. A method for fabricating an intracranial aneurysm microfluidic vascular chip as described in claim 1, characterized in that, Includes the following steps: A first module with the Y-shaped microfluidic channel was prepared using a mold; The opening is formed at the intersection of the Y-shaped microfluidic channel; Cover the opening with the elastic membrane; The preparation steps of the elastic membrane include: cleaning and sterilizing the polyisoprene film, followed by plasma treatment; A second module having the negative pressure suction cavity is prepared, wherein the negative pressure suction cavity has a first connection port communicating with the negative pressure suction cavity and a second connection port for connecting to a negative pressure suction device; The first connection port of the second module is connected to the opening of the first module, thereby connecting the second module to the first module, and the elastic membrane covers the opening and the first connection port.

7. The method for preparing the intracranial aneurysm microfluidic vascular chip according to claim 6, characterized in that, The plasma treatment is oxygen plasma treatment, and the treatment time is 1 to 3 minutes.

8. The method for preparing the intracranial aneurysm microfluidic vascular chip according to claim 7, characterized in that, After the polyisoprene film is subjected to plasma treatment, it is incubated for cell culture using fibronectin.

9. The application of an intracranial aneurysm microfluidic vascular chip as described in any one of claims 1 to 5, or an intracranial aneurysm microfluidic vascular chip prepared by the preparation method described in any one of claims 6 to 8, in intracranial aneurysm drug screening.

10. A microfluidic vascular chip system for intracranial aneurysms, characterized in that, The invention includes the intracranial aneurysm microfluidic vascular chip, circulation pump, reservoir, and negative pressure suction device as described in any one of claims 1 to 5. The negative pressure suction device is connected to the negative pressure suction cavity for evacuating the negative pressure suction cavity. The reservoir is connected to the main channel of the Y-shaped microfluidic channel through a first connecting tube. The reservoir is connected to the two branch channels of the Y-shaped microfluidic channel through a second connecting tube. The circulation pump is connected to the first connecting tube and / or the second connecting tube.

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