Multi-material bionic liver tissue organ chip capable of perfusion culture and preparation method thereof

Abstract

The present invention provides a multi-material bionic liver tissue organ chip capable of perfusion culture and a preparation method thereof. The multi-material bionic liver tissue organ chip comprises an upper vascular network module, a bionic liver tissue module, and a bottom vascular network trough module, which are sequentially connected. The upper vascular network module is provided with a perfusion inlet and a first bionic vascular network, and the bottom vascular network trough module is provided with a second bionic vascular network and a perfusion outlet. The bionic liver tissue module comprises an outer shell, within which are disposed N hepatic lobules, where N is greater than or equal to 3. Each hepatic lobule is provided with hepatic lobular connective tissue on the outer side, and a central hepatic lobule vein is provided running through the middle of each hepatic lobule from top to bottom. The first and second bionic vascular networks are provided with bionic central veins and bionic vascular connective tissue corresponding to the central hepatic lobule vein and hepatic lobular connective tissue of each hepatic lobule. The technical solution of the present invention has excellent biocompatibility and meets the requirements of perfusion culture.

Description

Technical Field

[0001] The present invention relates to the technical field of bioengineering, and in particular to a multi-material bionic liver tissue organ chip capable of perfusion culture and a preparation method thereof. Background Art

[0002] Currently, the fabrication and application of organ tissue chips primarily relies on PDMS materials and microfluidics. Traditional PDMS can absorb hydrophobic small molecules, such as biomolecules and drugs, from solutions. Many researchers have noted the adsorption of proteins on PDMS surfaces, which has become a major problem in molecular biology. For experiments investigating cell signaling and drug dose-response, the use of PDMS can significantly bias the final results.

[0003] Common materials used in biomimetic liver tissue 3D printing include biocompatible polymers such as PEGDA, GelMA, and HAMA. Printing techniques include extrusion, inkjet, and digital light processing. However, liver tissue has a complex physiological structure, and traditional single materials cannot accurately simulate the anisotropic physical properties of human liver lobule tissue. Numerous studies have demonstrated that differences in mechanical properties can significantly affect cell culture and the expression of various factors.

[0004] Furthermore, a reliable perfusion support vascular network is crucial for the survival of tissues and organs cultured in vitro. As the complexity of in vitro liver tissue increases, simple channels are no longer sufficient for the cultivation of complex tissues, necessitating the design of more complex perfusion channels. Summary of the Invention

[0005] In response to the above technical problems, the present invention discloses a multi-material bionic liver tissue organ chip capable of perfusion culture and a preparation method thereof.

[0006] To this end, the technical solution adopted in the present invention is:

[0007] A multi-material bionic liver tissue organ chip capable of perfusion culture comprises an upper vascular network module, a bionic liver tissue module, and a bottom vascular network tank module connected in sequence. The bottom vascular network tank module comprises a receiving tank for receiving the bionic liver tissue module and a bottom vascular network, wherein the bottom vascular network is located below the receiving tank.

[0008] The upper vascular network module is provided with a perfusion inlet and a first bionic vascular network, and the bottom vascular network tank module is provided with a second bionic vascular network and a perfusion outlet; the first bionic vascular network and the second bionic vascular network have the same structure;

[0009] The bionic liver tissue module includes a housing, wherein N hepatic lobules are disposed within the housing. The hepatic lobules are polygonal prisms and are interconnected along a circumferential direction to form a honeycomb shape, where N is greater than or equal to 3. Hepatic lobular connective tissue is disposed on the outside of each hepatic lobule, and a central hepatic lobular vein is disposed vertically through the middle of each hepatic lobule.

[0010] The first bionic vascular network and the second bionic vascular network are provided with a bionic vascular central vein and a bionic vascular connective tissue corresponding to the central vein of each hepatic lobule and the connective tissue of the hepatic lobule;

[0011] The outer shell, upper vascular network module, and lower vascular network tank module are molded using PEGDA bio-ink; the hepatic lobule connective tissue surrounding the hepatic lobule is molded using GelMA bio-ink, and the hepatic lobule is molded using side branch-modified GelMA bio-ink. The side branch-modified GelMA bio-ink is a bio-ink based on GelMA modification, and the modulus of the GelMA bio-ink and the side branch-modified GelMA bio-ink is substantially consistent with that of human connective tissue and hepatic lobule. The side branch-modified GelMA bio-ink can be made using existing technology.

[0012] The bottom of the upper vascular network module is provided with a connecting component, and the top of the bottom vascular network slot module is provided with a matching component. The connection between the upper vascular network module and the bottom vascular network slot module is achieved by connecting the connecting component and the matching component.

[0013] Using this technical solution, the outer shell, upper vascular network module, and bottom vascular network tank module are molded using PEGDA bio-ink. This material not only meets strength requirements but also provides clearance for cell growth and migration, and exhibits a certain degree of biocompatibility, providing strength support and improving structural stability for the organ-on-a-chip. The connective tissue and central veins are molded using GelMA bio-ink and collateral-modified GelMA bio-ink, which exhibit excellent biocompatibility. This allows the entire organ-on-a-chip to achieve enhanced biomimetic effects while meeting perfusion culture requirements.

[0014] If N is 3 to 6, the hepatic lobules are arranged circumferentially. If N>6, at least one hepatic lobule is located in the center, and the other hepatic lobules are arranged circumferentially around the center.

[0015] As a further improvement of the present invention, the PEGDA bio-ink comprises 15% w / v PEGDA, 0.25% w / v LAP, and 0.5% w / v sodium 2,2′-dihydroxy-4,4′-dimethoxybenzophenone 5,5′-disulfonate (R1800). Wherein, 1% w / v is 1 g / 100 ml, the same below.

[0016] As a further improvement of the present invention, the components of the GelMA bio-ink include GelMA 10% w / v, LAP 0.25% w / v, and R1800 0.5% w / v.

[0017] As a further improvement of the present invention, the components of the side-branch modified GelMA biological ink include side-branch modified GelMA 10% w / v, LAP 0.25% w / v, and R1800 0.5% w / v.

[0018] As a further improvement of the present invention, HUVEC cells are added to the GelMA bio-ink, and HepG2 cells are added to the side branch modified GelMA bio-ink. This technical solution has a better biomimetic effect.

[0019] As a further improvement of the present invention, the bionic liver tissue module is honeycomb-shaped; the diameter of the inscribed circle of the cross section of the liver lobule is 2-4 mm; and the thickness of the liver lobule is 1-3.5 mm. This technical solution is closer to human liver tissue.

[0020] As a further improvement of the present invention, the diameter of the central vein of the hepatic lobule is 0.5-1.1 mm, and the diameter of the capillaries of the connective tissue of the hepatic lobule is 0.15-0.4 mm.

[0021] As a further improvement of the present invention, the first bionic vascular network and the second bionic vascular network each comprise, from top to bottom, an inlet equalizing flow channel, an upstream bionic vascular network, a tissue flow channel, a downstream bionic vascular network, and an outlet collecting channel, which are sequentially connected; the upstream bionic vascular network and the downstream bionic vascular network have the same structure, and both correspond to N circumferentially arranged vascular network basic units of the liver lobule, and the N vascular network basic units are sequentially connected to form a network unit ring;

[0022] The basic unit of the vascular network includes a main channel, a first main branch, and two second main branches, wherein the first main branch and the two second main branches extend outward, the first main branch is located between the two second main branches, the upstream of the first main branch and the two second main branches are connected to one end of the main channel, the downstream of the two second main branches are respectively connected to the upstream of the first side branch and the second side branch, and the other end of the main channel is connected to the upstream of two third side branches, wherein the two third side branches are respectively located on both sides of the main channel;

[0023] At least one second side branch of the vascular network basic unit is connected to the second side branch of an adjacent vascular network basic unit, and a third side branch of the vascular network basic unit is connected to the third side branch of an adjacent vascular network basic unit; or the downstream of a third side branch of the vascular network basic unit is connected to the downstream of a second side branch of a vascular network basic unit on its right side, and the downstream of another third side branch of the vascular network basic unit is connected to the downstream of a third side branch of a vascular network basic unit on its right side;

[0024] The tissue flow-through channel includes a central vein and connective tissue flowing through the channel, the first main branch of the upstream bionic vascular network is connected to the first main branch of the downstream bionic vascular network via the central vein flowing through the channel, or the main channel and the first main branch of the upstream bionic vascular network are connected to the main channel and the first main branch of the downstream bionic vascular network respectively via the central vein flowing through the channel, and the first side branch / the second side branch / the third side branch of the upstream bionic vascular network are connected to the downstream bionic vascular network respectively via the connective tissue flowing through the channel;

[0025] The inlet equalizing channel is connected to the main channel of the upstream bionic blood vessel network, and the outlet converging channel is connected to the main channel of the downstream bionic blood vessel network.

[0026] This technical solution creates a hierarchical, bifurcated biomimetic vascular network that fully accounts for the complexity of tissues and organs, matching the structural and blood flow requirements of biomimetic liver tissue and simulating the blood flow distribution of central veins and connective tissue. 3D streamline simulations show that blood flow rates closely match those of human tissue, and the 3D flow path streamline distribution meets the requirements, ensuring that the fluid distribution is sufficient to supply the biomimetic liver tissue.

[0027] As a further improvement of the present invention, the first side branch, the second side branch and the second main branch are arranged in a Y shape; the two second main branches and the main channel are arranged in a Y shape;

[0028] The inlet equal flow channel and the outlet collecting channel both include a total channel and an equal flow channel, the total channel is connected to the equal flow channel, and the equal flow channel is connected to the main channel of each vascular network basic unit; the width of the total channel is greater than the width of the equal flow channel; the equal flow channel includes a circular channel and a circular central channel, the circular central channel is located in the middle of the circular channel, and its two ends are respectively connected to the circular channel; the equal flow channel is connected to the main channel of each vascular network basic unit through the inflow channel.

[0029] As a further improvement of the present invention, the inner diameter of the total channel is 0.8-1.5 mm, and the inner diameters of the flow-equalizing channel and the inflow channel are 0.35-0.55 times the inner diameter of the total channel; further, the inner diameters of the flow-equalizing channel and the inflow channel are 0.3-0.8 mm;

[0030] The inner diameter of the main channel is 0.5-0.9 mm, and the ratio of the thickness of the main channel to the inner diameter is 0.7-1.0;

[0031] The inner diameter of the first main branch is 0.7-0.88 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the first main branch is 0.85-1.2;

[0032] The inner diameter of the second main branch is 0.5-0.55 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the second main branch is 0.9-1.2;

[0033] The inner diameters of the first side branch, the second side branch, and the third side branch are 0.3-0.44 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the first side branch, the second side branch, and the third side branch is 0.9-1.6;

[0034] The angle between the third side branch and the main channel connected to it is 30°-60°, the angle between the first main branch and the main channel connected to it is 0°-10°, the angle between the first main branch and the second main branch connected to it is 20°-40°; the angle between the second main branch and the main channel connected to it is 130°-155°; the angle between the first side branch and the second main branch connected to it is 130°-170°, the angle between the second side branch and the first side branch connected to it is 65°-80°, and the angle between the second side branch and the second main branch connected to it is 125°-140°.

[0035] The inner diameter of the central vein flowing through the channel is 0.45-1.2 mm, and the inner diameter of the capillaries flowing through the connective tissue of the channel is 0.1-0.45 mm.

[0036] The vascular network required by the liver lobule can be adapted by changing the upstream bionic vascular network, tissue flow channels, downstream bionic vascular network array, and main and side branch parameters.

[0037] As a further improvement of the present invention, the upstream bionic vascular network and the downstream bionic vascular network each include a vertical portion of the vascular network connected to the tissue flow channel, the vertical portion of the vascular network includes a central vein of the vascular network and a connective tissue of the vascular network, the central vein of the vascular network is connected to the central vein of the flow channel, and the connective tissue of the vascular network is connected to the connective tissue of the flow channel;

[0038] The inner diameter of the central vein of the vascular network is 0.5-1.1 mm, and the inner diameter of the capillaries of the connective tissue of the vascular network is 0.15-0.4 mm; the inner diameter of the central vein of the channel is 0.85-1.15 times that of the central vein of the vascular network, and the inner diameter of the capillaries of the connective tissue of the channel is 0.9-1.1 times that of the capillaries of the connective tissue of the vascular network;

[0039] The inner diameter of the first main branch is 0.33-0.8 mm; the inner diameter of the second main branch is 0.28-0.5 mm; the inner diameters of the first side branch, the second side branch, and the third side branch are 0.15-0.4 mm.

[0040] As a further improvement of the present invention, when N>3, the vascular network basic unit further includes a third main branch, the main channel is connected to the upstream of the third main branch, the third side branches are located on both sides of the third main branch, the downstream of the third main branch of each vascular network basic unit is connected to the middle of the network unit ring, and the third main branch of the upstream bionic vascular network is connected to the downstream of the third main branch of the downstream bionic vascular network via a central vein flowing through the channel;

[0041] The inner diameter of the third main branch is 0.5-0.55 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the third main branch is 0.9-1.2;

[0042] The angle between the third main branch and the main channel connected thereto is 0°-10°, and the angle between the third side branch and the third main branch connected thereto is 120°-135°;

[0043] When N=3, the two third side branches of the basic unit of the vascular network are on a straight line.

[0044] The present invention also discloses a method for preparing the multi-material bionic liver tissue organ chip capable of perfusion culture as described above, comprising:

[0045] Step S1, using DLP printing to form an upper vascular network module, a bionic liver tissue module, and a bottom vascular network tank module;

[0046] Step S2: Place the bionic liver tissue module into the receiving slot of the bottom vascular network slot module, add PEGDA ink into the mating component, place the upper vascular network module on it, and connect the connecting component with the mating component, perform UV irradiation and post-curing to achieve connection and sealing.

[0047] Compared with the prior art, the present invention has the following beneficial effects:

[0048] The bionic liver tissue organ chip using the technical solution of the present invention has complex channels and has good mechanical properties while also having good biocompatibility, meeting the requirements of perfusion culture, better simulating the various properties of human liver tissue organs, and reducing the impact of cell culture and the expression of various factors. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1This is a schematic structural diagram of a multi-material bionic liver tissue organ chip capable of perfusion culture according to Example 1 of the present invention.

[0050] Figure 2 It is a structural diagram of the upper vascular network module of Example 1 of the present invention.

[0051] Figure 3 Schematic diagram of the structure of the bionic liver tissue module of Example 1 of the present invention.

[0052] Figure 4 It is a structural diagram of the bottom vascular network slot module of Example 1 of the present invention.

[0053] Figure 5 This is a perfusion image of the bionic liver tissue module according to Example 1 of the present invention.

[0054] Figure 6 Schematic diagram of the structure of the bionic vascular network of Example 2 of the present invention.

[0055] Figure 7 This is a top view of the bionic vascular network of Example 2 of the present invention.

[0056] Figure 8 This is a cross-sectional view of the bionic vascular network of Example 2 of the present invention.

[0057] Figure 9 3 is a streamline flow velocity diagram of the COMSOL simulation results of the bionic vascular network of Example 2 of the present invention.

[0058] Figure 10 This is a pressure distribution diagram of the COMSOL simulation results of the hierarchical bifurcated bionic vascular network for tissue perfusion culture according to Example 2 of the present invention.

[0059] Figure 11 This is a three-dimensional streamline flow velocity diagram of a three-dimensional network COMSOL simulation of a hierarchical bifurcated bionic vascular network for tissue perfusion culture according to Example 2 of the present invention.

[0060] Figure 12 This is an image of a hydrogel vascular network and corresponding perfusion channel printed using PEGDA according to Example 2 of the present invention, wherein (a) is the hydrogel vascular network and (b) is the perfusion channel image.

[0061] Figure 13 Schematic diagram of the structure of the bionic vascular network of Example 3 of the present invention.

[0062] Figure 14 This is a side view of the bionic vascular network of Example 3 of the present invention.

[0063] Figure 15This is a schematic structural diagram of a multi-material bionic liver tissue organ chip capable of perfusion culture according to Example 4 of the present invention.

[0064] Figure 16 4 is a top view of the bionic liver tissue organ chip according to Example 4 of the present invention.

[0065] Figure 17 Schematic diagram of the structure of the bionic vascular network of Example 4 of the present invention.

[0066] Figure 18 This is a top view of the bionic vascular network of Example 4 of the present invention.

[0067] Figure 19 3 is a streamline flow velocity diagram of the COMSOL simulation results of the bionic vascular network of Example 4 of the present invention.

[0068] Figure 20 This is a pressure distribution diagram of the COMSOL simulation results of the bionic vascular network of Example 4 of the present invention.

[0069] Figure 21 This is a three-dimensional streamline flow velocity diagram of the three-dimensional network COMSOL simulation of the bionic vascular network of Example 4 of the present invention.

[0070] Figure 22 Schematic diagram of the structure of the bionic vascular network of Example 5 of the present invention.

[0071] Figure 23 This is a side view of the bionic vascular network of Example 5 of the present invention.

[0072] Reference numerals include:

[0073] 1-upper vascular network module, 2-bionic liver tissue module, 3-bottom vascular network tank module;

[0074] 11- perfusion inlet, 12- first bionic vascular network, 13- connecting component;

[0075] 21-shell, 22-lobule, 23-lobular connective tissue, 24-central vein of the lobule;

[0076] 31 - perfusion outlet, 32 - second bionic vascular network, 33 - matching components;

[0077] 100-inlet equalizing channel, 200-upstream bionic vascular network, 300-tissue flow channel, 400-downstream bionic vascular network, 500-outlet collecting channel;

[0078] 110- total channel, 120- equal flow channel, 121- annular channel, 122- annular central channel, 123- inflow channel;

[0079] 210-basic unit of vascular network, 220-main channel, 230-first main branch, 240-second main branch, 250-first side branch, 260-second side branch, 270-third side branch, 280-third main branch;

[0080] 211-vertical part of the vascular network, 212-central vein of the vascular network, 213-connective tissue of the vascular network;

[0081] 310-flows through the central vein of the channel, 320-flows through the connective tissue of the channel. DETAILED DESCRIPTION

[0082] The preferred embodiments of the present invention are described in further detail below.

[0083] Example 1

[0084] like Figures 1 to 4 As shown, a multi-material bionic liver tissue organ chip capable of perfusion culture comprises an upper vascular network module 1, a bionic liver tissue module 2, and a bottom vascular network tank module 3 connected in sequence. The bottom vascular network tank module 3 comprises a receiving tank for receiving the bionic liver tissue module 2 and a bottom vascular network, wherein the bottom vascular network is located below the receiving tank.

[0085] The upper vascular network module 1 is provided with a perfusion inlet 11 and a first bionic vascular network 12, and the bottom vascular network tank module 3 is provided with a second bionic vascular network 32 and a perfusion outlet 31; the first bionic vascular network 12 and the second bionic vascular network 32 have the same structure;

[0086] The bionic liver tissue module 2 includes a housing 21, within which are disposed N hepatic lobules 22. Each hepatic lobule 22 is a polygonal prism and interconnected circumferentially. This embodiment illustrates seven hepatic lobules 22. Specifically, the seven hepatic lobules 22 are structured as follows: one hepatic lobule 22 is at the center, with six hepatic lobules 22 connected to the periphery, forming a honeycomb shape. Each hepatic lobule 22 is surrounded by hepatic connective tissue 23, and a central hepatic vein 24 runs through the center of each hepatic lobule 22.

[0087] The first bionic vascular network 12 and the second bionic vascular network 32 are provided with bionic vascular central veins and bionic vascular connective tissues corresponding to the hepatic lobule central vein 24 and the hepatic lobule connective tissue 23 of each hepatic lobule 22;

[0088] The bottom of the upper vascular network module 1 is provided with a connecting component 13, and the top of the bottom vascular network tank module 3 is provided with a mating component 33. The connection between the connecting component 13 and the mating component 33 is achieved by connecting the upper vascular network module 1 and the bottom vascular network tank module 3. Furthermore, the connecting component 13 is located around the bottom of the upper vascular network module 1, and the mating component 33 is located around the top of the bottom vascular network tank module 3, corresponding to the position of the connecting component 13.

[0089] The housing 21, upper vascular network module 1, and bottom vascular network tank module 3 are molded using PEGDA bio-ink; the hepatic lobule connective tissue surrounding the hepatic lobules is molded using GelMA bio-ink, and the hepatic lobules are molded using side branch-modified GelMA bio-ink. Specifically, the PEGDA bio-ink comprises 15% w / v PEGDA, 0.25% w / v LAP, and 0.5% w / v sodium 2,2′-dihydroxy-4,4′-dimethoxybenzophenone 5,5′-disulfonate (R1800). The GelMA bio-ink comprises 10% w / v GelMA, 0.25% w / v LAP, and 0.5% w / v R1800. The side branch-modified GelMA bio-ink comprises 10% w / v side branch-modified GelMA, 0.25% w / v LAP, and 0.5% w / v R1800. HUVEC cells are added to the GelMA bio-ink, and HepG2 cells are added to the side-branch modified GelMA bio-ink. The side-branch modified GelMA uses materials from the prior art.

[0090] The bionic liver tissue module 2 is honeycomb-shaped. Furthermore, the diameter of the inscribed circle of the cross section of the hepatic lobule 22 is 2-4 mm, and the thickness of the hepatic lobule 22 is 1-3.5 mm. The diameter of the central vein 24 of the hepatic lobule is 0.5-1.1 mm, and the diameter of the capillaries of the hepatic lobule connective tissue 23 is 0.15-0.4 mm.

[0091] The method for preparing the multi-material bionic liver tissue 2 organ chip capable of perfusion culture comprises:

[0092] Step S1, using DLP to print and form the upper vascular network module 1, the bionic liver tissue module 2, and the bottom vascular network slot module 3 respectively;

[0093] Among them, the bionic liver tissue module 2 is prepared by multi-material printing. The ink configuration has been described above and is prepared using a DLP multi-material printing device. The three materials are printed layer by layer. Different materials are switched by moving the liquid pool platform horizontally. When switching different materials, heated deionized water is used for cleaning as needed, that is, the printing platform is lifted vertically from the liquid pool, and the liquid pool platform is moved horizontally to the top of the cleaning nozzle (fixed in the center of the waste liquid pool). Deionized water is sprayed for flushing, and then hot air is blown to promote the evaporation of deionized water. Finally, the liquid pool platform moves back to the top of the liquid pool of the corresponding material for the next solidification. The vertical direction controls the thickness of the single-layer solidification by controlling the distance between the printing platform and the bottom of the liquid pool. The bionic liver tissue module 2 under the electron microscope is shown Figure 5 shown.

[0094] In step S2, the bionic liver tissue module 2 is loaded into the receiving slot of the bottom vascular network slot module 3, PEGDA ink is added to the matching component 33, the upper vascular network module 1 is placed thereon, and the connecting component 13 is matched and connected with the matching component 33, and then cured by UV irradiation to achieve connection and sealing.

[0095] After the cell-laden bio-ink is prepared and the entire structure is printed and assembled, it is immersed in culture medium. Simultaneously, a peristaltic pump is used to perfuse a 1:1 mixture of HepG2 and HUVEC culture medium through the inlet for incubation, and the perfused liquid is withdrawn through the outlet. To form a vascular channel through the perfusion channel, a fibrin solution is perfused after assembly. The inlet and outlet are sealed and allowed to stand for a period of time before the solution is withdrawn. The HUVEC solution is then perfused. The inlet and outlet are sealed and allowed to stand for a period of time to allow HUVECs to adhere to the wall and grow. Finally, the perfusion culture is continued in culture medium.

[0096] Example 2

[0097] like Figures 6 to 12 As shown, based on Example 1, in this embodiment, the first bionic vascular network 12 and the second bionic vascular network 32 have the same structure, and both include an inlet equalizing channel 100, an upstream bionic vascular network 200, a tissue flow channel 300, a downstream bionic vascular network 400 and an outlet collecting channel 500 connected in sequence from top to bottom. The upstream bionic vascular network 200 and the downstream bionic vascular network 400 have the same structure, and both correspond to 6 vascular network basic units 210 symmetrically arranged along the circumference, corresponding to the six hepatic lobules 22 around the bionic liver tissue module 2. The 6 vascular network basic units 210 are sequentially connected in the circumferential direction to form a network unit ring.

[0098] The vascular network basic unit 210 includes a main channel 220, a first main branch 230, two second main branches 240, and a third main branch 280. The first main branch 230 and the two second main branches 240 extend radially outward, with the first main branch 230 located between the two second main branches 240. The upstream ends of the first main branch 230 and the two second main branches 240 are connected to one end of the main channel 220, and the downstream ends of the two second main branches 240 are connected to the upstream ends of the first side branch 250 and the second side branch 260, respectively. The other end of the main channel 220 is connected to the upstream ends of the third main branch 280 and the upstream ends of the two third side branches 270. The two third side branches 270 are located on either side of the main channel 220 and the third main branch 280, respectively. The downstream of the third main branch 280 of each vascular network basic unit 210 is connected to the middle of the network unit ring, and the downstream of the third main branch 280 of the upstream bionic vascular network 200 is connected to the downstream of the third main branch 280 of the downstream bionic vascular network 400 through the central vein 310 flowing through the channel.

[0099] A second side branch 260 of the vascular network basic unit 210 is connected to the second side branch 260 of the adjacent vascular network basic unit 210. The downstream of the third side branch 270 of the vascular network basic unit 210 is connected to the downstream of the third side branch 270 of the adjacent vascular network basic unit 210, forming a hexagonal star.

[0100] The tissue flow-through channel 300 includes a flow-through channel central vein 310 and a flow-through channel connective tissue 320. The upstream bionic vascular network 200 is connected to the first main branch 230 of the downstream bionic vascular network 400 through the flow-through channel central vein, or the main channel 220 and the first main branch 230 of the upstream bionic vascular network 200 are connected to the main channel 220 and the first main branch 230 of the downstream bionic vascular network 400 through the flow-through channel central vein 310, and the first side branch 250 / the second side branch 260 / the third side branch 270 of the upstream bionic vascular network 200 and the downstream bionic vascular network 400 are connected respectively through the flow-through channel connective tissue 320.

[0101] The inlet equalizing channel 100 is connected to the main channel 220 of the upstream bionic vascular network 200 , and the outlet converging channel 500 is connected to the main channel 220 of the downstream bionic vascular network 400 .

[0102] The first side branch 250 , the second side branch 260 and the second main branch 240 are arranged in a Y shape. The two second main branches 240 and the main channel 220 are arranged in a Y shape. The connection between the main channel 220 and the third side branch 270 corresponds to the annular channel 121 .

[0103] The inlet equalizing flow channel 100 and the outlet converging channel 500 both include a main channel 110 and an equalizing flow channel 120. The main channel 110 is in communication with the equalizing flow channel 120, which is connected to the main channel 220 of each vascular network basic unit 210. The width of the main channel 110 is greater than that of the equalizing flow channel 120. The equalizing flow channel 120 includes an annular channel 121 and an annular central channel 122. The annular central channel is located in the middle of the annular channel 121, with both ends connected to the annular channel 121. The equalizing flow channel 120 is connected to the main channel 220 of each vascular network basic unit 210 via an inflow channel 123.

[0104] Furthermore, as shown in Table 1, the inner diameter of the total channel 110 is 0.8-1.5 mm, the inner diameters of the flow-equalizing channel 120 and the inflow channel 123 are 0.35-0.55 times the inner diameter of the total channel 110; and the inner diameters of the flow-equalizing channel 120 and the inflow channel 123 are 0.3-0.8 mm.

[0105] Table 1

[0106]

[0107] Furthermore, as shown in Table 2, the inner diameter of the main channel 220 is 0.5-0.9 mm, and the ratio of the thickness of the main channel 220 to the inner diameter is 0.7-1.0; the inner diameter of the first main branch 230 is 0.7-0.88 times the inner diameter of the main channel 220, and the ratio of the thickness of the first main branch 230 to the inner diameter is 0.85-1.2; the inner diameter of the second main branch 240 is 0.5-0.55 times the inner diameter of the main channel 220, and the ratio of the thickness of the second main branch 240 to the inner diameter is 0.9-1.2; the inner diameter of the third main branch 280 is 0.5-0.55 times the inner diameter of the main channel 220, and the ratio of the thickness of the third main branch 280 to the inner diameter is 0.9-1.2.

[0108] The inner diameters of the first, second, and third side branches 250, 260, and 270 are 0.3-0.44 times the inner diameter of the main channel 220, and the ratio of thickness to inner diameter of the first, second, and third side branches 250, 260, and 270 is 0.9-1.6. The inner diameters of the first main branch 230 are 0.33-0.8 mm; the inner diameters of the second main branch 240 are 0.28-0.5 mm; and the inner diameters of the first, second, and third side branches 250, 260, and 270 are 0.15-0.4 mm. The angle between the third side branch 270 and the main channel 220 connected thereto is 30°-60°. The angle between the first main branch 230 and the main channel 220 connected thereto is 0°-10°. The angle between the first main branch 230 and the second main branch 240 connected thereto is 20°-40°. The angle between the second main branch 240 and the main channel 220 connected thereto is 130°-155°. The angle between the third main branch 280 and the main channel 220 connected thereto is 0°-10°. The angle between the first side branch 250 and the second main branch 240 connected thereto is 130°-170°. The angle between the second side branch 260 and the first side branch 250 connected thereto is 65°-80°. The angle between the second side branch 260 and the second main branch 240 connected thereto is 125°-140°. The included angle between the third side branch 270 and the third main branch 280 connected thereto is 120°-135°.

[0109] Table 2

[0110]

[0111] Furthermore, the inner diameter of the central vein 310 flowing through the channel is 0.45-1.2 mm, and the inner diameter of the capillaries flowing through the connective tissue 320 of the channel is 0.1-0.45 mm.

[0112] The vascular network was simulated and verified using COMSOL software. The simulation parameters were as follows: water as the simulated fluid, an inlet velocity of 0.15 m / s, and an outlet set to atmospheric pressure. The inlet position was arranged according to the inlet flow channel 100.

[0113] The results are as follows Figure 9 and Figure 10 As shown in the figure, it can be seen that the flow velocity of the planar vascular network is very close to that of human tissue. The fluid distribution meets the blood flow distribution of the central vein and connective tissue of the organ tissue, and the pressure distribution is uniform, indicating that the flow resistance is low. Next, a three-dimensional streamline simulation is performed, and the results are shown in the figure. Figure 11 As shown, it can be seen that the streamline distribution of the three-dimensional flow channel meets the requirements and the fluid distribution can supply the corresponding bionic tissue.

[0114] The above-mentioned bionic vascular network adopts photo-crosslinked biological ink and is formed in one step by DLP printing. The photo-crosslinked biological ink can be selected from PEGDA ink. Specifically, the preparation method includes: after slicing the model of the hierarchical and bifurcated bionic vascular network for tissue perfusion culture, each layer is printed in sequence using photo-crosslinked biological ink, and the biological ink is photo-crosslinked using a UV projector to stack layer by layer. In order to generate an unobstructed vascular network, a cleaning step is required after each layer is printed. When one layer is cured, the printing platform is lifted vertically from the liquid pool, and then the liquid pool platform is translated to align the printing structure with the cleaning nozzle. The nozzle sprays deionized water to rinse away the residual PEGDA ink in the flow channel to prevent the channel from being blocked by curing after UV irradiation when the next layer is printed. After the flushing is completed, the remaining deionized water is sucked away from the side edge with a special dust-free paper towel to prevent the residual deionized water from locally diluting the biological ink and affecting the curing of the lower layer. The hydrogel vascular network printed with PEGDA and the perfusion experiment on the hydrogel vascular network are shown as follows: Figure 12 As shown, the results are consistent with the simulation.

[0115] Example 3

[0116] like Figure 13 and Figure 14 As shown, based on Example 1, in this embodiment, the upstream bionic vascular network 200 and the downstream bionic vascular network 400 include a vascular network vertical portion 211 connected to the tissue flow channel 300, and the vascular network vertical portion 211 includes a vascular network central vein 212 and a vascular network connective tissue 213, the vascular network central vein 212 is connected to the central vein 310 flowing through the channel, and the vascular network connective tissue 213 is connected to the connective tissue 320 flowing through the channel.

[0117] As shown in Table 3, the inner diameter of the central vein 212 of the vascular network is 0.5-1.1 mm, and the inner diameter of the capillaries of the vascular network connective tissue 213 is 0.15-0.4 mm; the inner diameter of the capillaries of the vascular network connective tissue 213 is 0.3-0.5 times the inner diameter of the central vein 212 of the vascular network. As shown in Table 4, the inner diameter of the central vein 310 of the flow channel is 0.45-1.2 mm, and the inner diameter of the capillaries of the flow channel connective tissue 320 is 0.1-0.45 mm. In this embodiment, the inner diameter of the central vein 310 of the flow channel is 0.85-1.0 times the inner diameter of the central vein 212 of the vascular network, and the inner diameter of the capillaries of the flow channel connective tissue 320 is 0.9-1.0 times the inner diameter of the capillaries of the vascular network connective tissue 213. That is, the inner diameter of the central vein 310 flowing through the channel is larger than the inner diameter of the central vein 212 of the vascular network, and the inner diameter of the capillaries flowing through the connective tissue 320 of the channel is smaller than the inner diameter of the capillaries of the connective tissue 213 of the vascular network.

[0118] Table 3

[0119]

[0120] Table 4

[0121]

[0122] Example 4

[0123] like Figure 15 and Figure 16 As shown, based on Example 1, this embodiment is different in that the bionic liver tissue module 2 is composed of three liver lobules 22, which are sequentially connected along the circumferential direction. Other structures and materials are the same as those in Example 1.

[0124] Correspondingly, the first bionic vascular network 12 and the second bionic vascular network 32 also correspond to the three hepatic lobules 22. Figure 17 and Figure 18 As shown, the first bionic vascular network 12 and the second bionic vascular network 32 have the same structure, and both include, from top to bottom, an inlet equalizing flow channel 100, an upstream bionic vascular network 200, a tissue flow channel 300, a downstream bionic vascular network 400 and an outlet collecting channel 500 connected in sequence. The upstream bionic vascular network 200 and the downstream bionic vascular network 400 have the same structure, and both include three circumferentially symmetrically arranged vascular network basic units 210, which correspond to the three hepatic lobules 22 of the bionic liver tissue module 2 respectively.

[0125] The vascular network basic unit 210 includes a main channel 220, a first main branch 230, and two second main branches 240. The first main branch 230 and the two second main branches 240 extend radially outward, and the first main branch 230 is located between the two second main branches 240. The upstream of the first main branch 230 and the two second main branches 240 are connected to one end of the main channel 220, and the downstream of the two second main branches 240 are respectively connected to the upstream of the first side branch 250 and the second side branch 260. The other end of the main channel 220 is connected to the upstream of two third side branches 270. The two third side branches 270 are respectively located on both sides of the main channel 220.

[0126] The downstream of a third side branch 270 of the vascular network basic unit 210 is connected to the downstream of a second side branch 260 of the right vascular network basic unit 210, and the downstream of another third side branch 270 of the vascular network basic unit 210 is connected to the downstream of a third side branch 270 of the right vascular network basic unit 210.

[0127] The tissue flow-through channel 300 includes a flow-through channel central vein 310 and a flow-through channel connective tissue 320. The first main branch 230 of the upstream bionic vascular network 200 is connected to the first main branch 230 of the downstream bionic vascular network 400 via the flow-through channel central vein, or the main channel 220 and the first main branch 230 of the upstream bionic vascular network 200 are connected to the main channel 220 and the first main branch 230 of the downstream bionic vascular network 400 via the flow-through channel central vein 310, and the first side branch 250 / the second side branch 260 / the third side branch 270 of the upstream bionic vascular network 200 and the downstream bionic vascular network 400 are connected respectively via the flow-through channel connective tissue 320.

[0128] The inlet equalizing channel 100 is connected to the main channel 220 of the upstream bionic vascular network 200 , and the outlet converging channel 500 is connected to the main channel 220 of the downstream bionic vascular network 400 .

[0129] The first side branch 250 , the second side branch 260 and the second main branch 240 are arranged in a Y shape. The two second main branches 240 and the main channel 220 are arranged in a Y shape. The connection between the main channel 220 and the third side branch 270 corresponds to the annular channel 121 .

[0130] The inlet equalizing flow channel 100 and the outlet converging channel 500 both include a main channel 110 and an equalizing flow channel 120. The main channel 110 is in communication with the equalizing flow channel 120, which is connected to the main channel 220 of each vascular network basic unit 210. The width of the main channel 110 is greater than that of the equalizing flow channel 120. The equalizing flow channel 120 includes an annular channel 121 and an annular central channel 122. The annular central channel is located in the middle of the annular channel 121, with both ends connected to the annular channel 121. The equalizing flow channel 120 is connected to the main channel 220 of each vascular network basic unit 210 via an inflow channel 123.

[0131] The dimensional requirements for the main channel 110, annular channel 121, annular central channel 122, and inflow channel 123 are shown in Table 1. The dimensional and angular requirements for the main channel 220, first main branch 230, second main branch 240, first side branch 250, and second side branch 260 are shown in Table 2. The two third side branches 270 of the vascular network basic unit 210 are aligned. The dimensions of the third side branches 270 are shown in Table 2.

[0132] The inner diameter of the central vein 310 flowing through the channel is 0.45-1.2 mm, and the inner diameter of the capillaries flowing through the connective tissue 320 of the channel is 0.1-0.45 mm.

[0133] The preparation methods of the first bionic vascular network 12 and the second bionic vascular network 32 in this embodiment are the same as those in Example 2.

[0134] The vascular network was simulated and verified using COMSOL software. The simulation parameters were as follows: water as the simulated fluid, an inlet velocity of 0.15 m / s, and an outlet set to atmospheric pressure. The inlet position was arranged according to the inlet flow channel 100.

[0135] The results are as follows Figure 19 and Figure 20 As shown in the figure, it can be seen that the flow velocity of the planar vascular network is very close to that of human tissue. The fluid distribution meets the blood flow distribution of the central vein and connective tissue of the organ tissue, and the pressure distribution is uniform, indicating that the flow resistance is low. Next, a three-dimensional streamline simulation is performed, and the results are shown in the figure. Figure 21 As shown, it can be seen that the streamline distribution of the three-dimensional flow channel meets the requirements and the fluid distribution can supply the corresponding bionic tissue.

[0136] Example 5

[0137] like Figure 22 and Figure 23 As shown, based on Example 4, in this embodiment, the upstream bionic vascular network 200 and the downstream bionic vascular network 400 include a vascular network vertical portion 211 connected to the tissue flow channel 300, and the vascular network vertical portion 211 includes a vascular network central vein 212 and a vascular network connective tissue 213, the vascular network central vein 212 is connected to the central vein 310 flowing through the channel, and the vascular network connective tissue 213 is connected to the connective tissue 320 flowing through the channel.

[0138] As shown in Table 3, the inner diameter of the central vein 212 of the vascular network is 0.5-1.1 mm, and the inner diameter of the connective tissue 213 of the vascular network is 0.15-0.4 mm; the inner diameter of the connective tissue 213 of the vascular network is 0.3-0.5 times the inner diameter of the central vein 212 of the vascular network. As shown in Table 4, the inner diameter of the central vein 310 of the flow channel is 0.45-1.2 mm, and the inner diameter of the connective tissue 320 of the flow channel is 0.1-0.45 mm. In this embodiment, the inner diameter of the central vein 310 of the flow channel is 0.85-1.0 times that of the central vein 212 of the vascular network, and the inner diameter of the capillaries of the connective tissue 320 of the flow channel is 0.9-1.0 times the inner diameter of the capillaries of the connective tissue 213 of the vascular network. That is, the inner diameter of the central vein 310 flowing through the channel is larger than the inner diameter of the central vein 212 of the vascular network, and the inner diameter of the capillaries flowing through the connective tissue 320 of the channel is smaller than the inner diameter of the capillaries of the connective tissue 213 of the vascular network.

[0139] In the description of the present invention, it should be understood that terms such as "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating orientations or positional relationships, are orientations or positional relationships based on the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0140] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of the technical features being referred to. Thus, a feature identified as "first" or "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, "plurality" means two or more, unless otherwise specifically defined.

[0141] In the present invention, unless otherwise expressly specified or limited, terms such as "mounted," "connected," "connect," and "fixed" should be understood broadly. For example, they may refer to fixed connection, detachable connection, or integration; they may refer to direct connection or indirect connection through an intermediate medium; they may refer to internal communication between two components or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0142] The above is a further detailed description of the present invention in conjunction with specific preferred embodiments, and the specific implementation of the present invention should not be considered to be limited to these descriptions. For those skilled in the art of the present invention, without departing from the concept of the present invention, several simple deductions or substitutions can be made, which should be considered to fall within the scope of protection of the present invention.

Claims

1. A multi-material bionic liver tissue organ chip capable of perfusion culture, characterized by: The device comprises an upper vascular network module, a bionic liver tissue module, and a bottom vascular network tank module connected in sequence. The bottom vascular network tank module comprises a receiving tank for receiving the bionic liver tissue module and a bottom vascular network. The bottom vascular network is located below the receiving tank. The upper vascular network module is provided with a perfusion inlet and a first bionic vascular network, and the bottom vascular network tank module is provided with a second bionic vascular network and a perfusion outlet; the first bionic vascular network and the second bionic vascular network have the same structure; The bionic liver tissue module includes a housing, wherein N hepatic lobules are disposed within the housing. The hepatic lobules are hexagonal prisms and are interconnected along a circumferential direction, wherein N is greater than or equal to 3. Hepatic lobular connective tissue is disposed on the outside of each hepatic lobule, and a central vein of the hepatic lobule is disposed vertically through the middle of each hepatic lobule. The first bionic vascular network and the second bionic vascular network are provided with a bionic vascular central vein and a bionic vascular connective tissue corresponding to the central vein of each hepatic lobule and the connective tissue of the hepatic lobule; The shell, the upper vascular network module, and the bottom vascular network tank module are formed by using PEGDA bio-ink; the hepatic lobule connective tissue surrounding the hepatic lobule is formed by using GelMA bio-ink, and the hepatic lobule is formed by using side branch modified GelMA bio-ink; The bottom of the upper vascular network module is provided with a connecting component, and the top of the bottom vascular network slot module is provided with a matching component. The connection between the upper vascular network module and the bottom vascular network slot module is achieved by connecting the connecting component and the matching component.

2. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 1, characterized in that: The components of the PEGDA bio-ink include PEGDA 15% w / v, LAP 0.25% w / v, and 2,2′-dihydroxy-4,4′-dimethoxybenzophenone 5,5′-disulfonic acid sodium 0.5% w / v; The components of the GelMA bio-ink include GelMA 10% w / v, LAP 0.25% w / v, and R1800 0.5% w / v; the components of the side-branch modified GelMA bio-ink include side-branch modified GelMA-10% w / v, LAP 0.25% w / v, and R18000.5% w / v.

3. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 2, characterized in that: HUVEC cells are added to the GelMA bio-ink, and HepG2 cells are added to the side branch modified GelMA bio-ink.

4. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 2, characterized in that: The bionic liver tissue module is honeycomb-shaped; the diameter of the inscribed circle of the cross section of the liver lobule is 2-4 mm; and the thickness of the liver lobule is 1-3.5 mm.

5. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 4, characterized in that: The diameter of the central vein of the hepatic lobule is 0.5-1.1 mm, and the diameter of the capillaries of the connective tissue of the hepatic lobule is 0.15-0.4 mm.

6. The multi-material bionic liver tissue organ chip capable of perfusion culture according to any one of claims 1 to 5, characterized in that: The first bionic vascular network and the second bionic vascular network both include, from top to bottom, an inlet equalizing flow channel, an upstream bionic vascular network, a tissue flow channel, a downstream bionic vascular network and an outlet collecting channel, which are connected in sequence. The upstream bionic vascular network and the downstream bionic vascular network have the same structure, and both include a plurality of vascular network basic units arranged circumferentially and corresponding to the liver lobules, and the plurality of vascular network basic units are sequentially connected to form a network unit ring; the vascular network basic unit includes a main channel, a first main branch and two second main branches, the first main branch and the two second main branches extend outward, the first main branch is located between the two second main branches, the upstream of the first main branch and the two second main branches are connected to one end of the main channel, the downstream of the two second main branches are respectively connected to the upstream of the first side branch and the second side branch, the other end of the main channel is connected to the upstream of two third side branches, and the two third side branches are respectively located on both sides of the main channel; At least one second side branch of the vascular network basic unit is connected to the second side branch of an adjacent vascular network basic unit, and a third side branch of the vascular network basic unit is connected to the third side branch of an adjacent vascular network basic unit; or the downstream of a third side branch of the vascular network basic unit is connected to the downstream of a second side branch of a vascular network basic unit on its right side, and the downstream of another third side branch of the vascular network basic unit is connected to the downstream of a third side branch of a vascular network basic unit on its right side; The tissue flow-through channel includes a central vein and connective tissue flowing through the channel, the first main branch of the upstream bionic vascular network is connected to the first main branch of the downstream bionic vascular network via the central vein flowing through the channel, or the main channel and the first main branch of the upstream bionic vascular network are connected to the main channel and the first main branch of the downstream bionic vascular network respectively via the central vein flowing through the channel, and the first side branch / the second side branch / the third side branch of the upstream bionic vascular network are connected to the downstream bionic vascular network respectively via the connective tissue flowing through the channel; The inlet equalizing channel is connected to the main channel of the upstream bionic blood vessel network, and the outlet converging channel is connected to the main channel of the downstream bionic blood vessel network.

7. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 6, characterized in that: The first side branch, the second side branch and the second main branch are arranged in a Y shape; the two second main branches and the main channel are arranged in a Y shape; The inlet equal flow channel and the outlet collecting channel both include a total channel and an equal flow channel, the total channel is connected to the equal flow channel, and the equal flow channel is connected to the main channel of each vascular network basic unit; the width of the total channel is greater than the width of the equal flow channel; the equal flow channel includes a circular channel and a circular central channel, the circular central channel is located in the middle of the circular channel, and its two ends are respectively connected to the circular channel; the equal flow channel is connected to the main channel of each vascular network basic unit through the inflow channel.

8. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 7, characterized in that: The inner diameter of the total channel is 0.8-1.5 mm, and the inner diameters of the flow-equalizing channel and the inflow channel are 0.35-0.55 times the inner diameter of the total channel; The inner diameter of the main channel is 0.5-0.9 mm, and the ratio of the thickness to the inner diameter of the main channel is 0.7-1.0; The inner diameter of the first main branch is 0.7-0.88 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the first main branch is 0.85-1.2; The inner diameter of the second main branch is 0.5-0.55 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the second main branch is 0.9-1.2; The inner diameters of the first side branch, the second side branch, and the third side branch are 0.3-0.44 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the first side branch, the second side branch, and the third side branch is 0.9-1.6; The angle between the third side branch and the main channel connected to it is 30°-60°, the angle between the first main branch and the main channel connected to it is 0°-10°, the angle between the first main branch and the second main branch connected to it is 20°-40°; the angle between the second main branch and the main channel connected to it is 130°-155°; the angle between the first side branch and the second main branch connected to it is 130°-170°, the angle between the second side branch and the first side branch connected to it is 65°-80°, and the angle between the second side branch and the second main branch connected to it is 125°-140°. The inner diameter of the central vein flowing through the channel is 0.45-1.2 mm, and the inner diameter of the capillaries flowing through the connective tissue of the channel is 0.1-0.45 mm.

9. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 8, characterized in that: The upstream bionic vascular network and the downstream bionic vascular network each include a vertical portion of the vascular network connected to the tissue flow channel, the vertical portion of the vascular network includes a central vein of the vascular network and a connective tissue of the vascular network, the central vein of the vascular network is connected to the central vein of the flow channel, and the connective tissue of the vascular network is connected to the connective tissue of the flow channel; The inner diameter of the central vein of the vascular network is 0.5-1.1 mm, and the inner diameter of the connective tissue of the vascular network is 0.15-0.4 mm; the inner diameter of the central vein flowing through the channel is 0.85-1.15 times that of the central vein of the vascular network, and the inner diameter of the capillaries of the connective tissue flowing through the channel is 0.9-1.1 times that of the capillaries of the connective tissue of the vascular network; The inner diameter of the first main branch is 0.33-0.8 mm; the inner diameter of the second main branch is 0.28-0.5 mm; the inner diameters of the first side branch, the second side branch, and the third side branch are 0.15-0.4 mm.

10. The multi-material bionic liver tissue organ chip capable of perfusion culture according to claim 9, characterized in that: When N>6, the vascular network basic unit further includes a third main branch, the main channel is connected to the upstream of the third main branch, the third side branches are located on both sides of the third main branch, the downstream of the third main branch of each vascular network basic unit is connected to the middle of the network unit ring, and the third main branch of the upstream bionic vascular network is connected to the downstream of the third main branch of the downstream bionic vascular network through the central vein of the flow channel; The inner diameter of the third main branch is 0.5-0.55 times the inner diameter of the main channel, and the ratio of the thickness to the inner diameter of the third main branch is 0.9-1.2; The angle between the third main branch and the main channel connected thereto is 0°-10°, and the angle between the third side branch and the third main branch connected thereto is 120°-135°; When N=3, the two third side branches of the basic unit of the vascular network are on a straight line.

11. The method for preparing a multi-material bionic liver tissue organ chip capable of perfusion culture according to any one of claims 1 to 10, characterized in that: include: Step S1, using DLP printing to form an upper vascular network module, a bionic liver tissue module, and a bottom vascular network tank module; Step S2: Place the bionic liver tissue module into the receiving slot of the bottom vascular network slot module, add PEGDA ink into the mating component, place the upper vascular network module on it, and connect the connecting component with the mating component, perform UV irradiation and post-curing to achieve connection and sealing.

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