Biological 3D printing visual craniocerebral tumor drug in-vitro screening model device and application thereof

Abstract

The invention belongs to the technical field of biological equipment, and particularly relates to a biological 3D printing visual craniocerebral tumor drug in-vitro screening model device and application thereof. The visual craniocerebral tumor drug in-vitro screening model device is mainly composed of a liquid inlet, a three-way valve, a bionic blood brain barrier model, a tumor model culture area, a culture container, a container cover, a cell culture bag, a camera and a cell activity detection electrode. The bionic blood brain barrier model and the tumor model can be realized by adopting a biological 3D printing mode; the construction of the in-vitro bionic blood-brain barrier model and the construction of the tumor model are integrated, and the method can be used for assisting in evaluating the type of the craniocerebral tumor treatment medicine and screening and evaluating the dosage of the medicine. Visualization of dynamic progress and drug screening in the rapid growth process of tumor cells is achieved, a stable and high-throughput drug screening chip is preliminarily formed through real-time monitoring of the cell activity detection electrode, and an auxiliary basis is provided for clinical treatment research of craniocerebral tumors.

Description

Technical Field

[0001] This invention belongs to the field of bio-device technology, specifically relating to a bio-3D printed visualization in vitro screening model device for intracranial tumor drugs and its application. Background Technology

[0002] Intracranial tumors can severely impact a patient's neurological function, especially malignant brain tumors which grow rapidly. Although various surgical, radiotherapy, chemotherapy, targeted therapy, and immunotherapy techniques are gradually developing, they have not yet been effectively controlled. The unique blood-brain / tumor barrier within the brain significantly limits the ability of chemotherapeutic drugs and cellular agents to penetrate intracranial tumor tissue. While there are numerous reports on the construction of biomimetic blood-brain barrier models, in vitro construction of biomimetic blood-brain / tumor barrier models is limited, especially given the rapid growth of tumors and the need for real-time monitoring of their progression. Therefore, there is a need to develop a visual in vitro drug screening model device for intracranial tumors. Summary of the Invention

[0003] To address the aforementioned technical issues, this invention provides a bio-3D printed visualization model device for in vitro screening of intracranial tumor drugs and its application. This device can simulate the interaction between the blood-brain barrier model and intracranial tumors, facilitating the research and development of intracranial tumor drug screening.

[0004] A bio-3D printed visualization model device for in vitro screening of intracranial tumor drugs includes: Liquid inlet system: Equipped with an inlet and a three-way valve for injecting culture medium, drugs, or cell suspensions; Biomimetic blood-brain barrier model: Constructed layer by layer using bio-3D printing technology, including a layer of brain microvascular endothelial cells, a layer of pericytes, and astrocytes; Tumor model culture area: Contains cell culture bags for culturing tumor cells, tissues, or organoids, equipped with a camera and cell viability detection electrodes; Culture container: Equipped with left and right interfaces connecting to the system piping, the container lid is a detachable screw cap, and it is equipped with a breathable filter membrane and electrode / camera sockets; Liquid outlet system: Equipped with a three-way valve and a liquid outlet for sampling or draining; The various parts of the device are connected by pipelines to form a closed-loop or unidirectional flow system.

[0005] Furthermore, the cell culture bag is equipped with a filter mesh to prevent cells from escaping; The camera is connected to a high-definition recording system to enable real-time dynamic observation; The external port of the three-way valve is equipped with a heparin cap for easy injection or sampling.

[0006] The beneficial effects that can be achieved are as follows: Integrating blood-brain barrier and tumor models, it more realistically simulates the intracranial drug delivery process; Supports multiple screening methods, including drug permeability, dose-effect, and immune cell delivery; It features real-time monitoring and visualization capabilities, improving screening efficiency and data reliability; The modular structure facilitates high-throughput expansion and standardized operation.

[0007] Furthermore, the present invention provides a bio-3D printed visualization in vitro screening model device for intracranial tumor drugs, including a system pipeline, wherein the system pipeline includes a liquid inlet, a three-way valve, a biomimetic blood-brain barrier model, a tumor model culture area, a cell culture bag, a container cap, a cell viability detection electrode, a camera, a three-way valve, and a liquid outlet. Both the inlet and outlet are equipped with removable sealing caps; Both the inlet and outlet are equipped with bacterial filter membranes. The system pipeline is equipped with two three-way valves at the head end and the tail end; The three-way valve can be used with a syringe to inject or extract liquid. The liquid injection can be one or more of the following: drug solution, cell solution, and nutrient solution; All external ports of the three-way valve are equipped with removable sealing caps. The sealing cap is equipped with a heparin cap; Both the biomimetic blood-brain barrier model and the tumor organoid model are achieved through ordered printing using bio-3D printing. The printing materials involved in the bio-3D printing method can be one or more of the following: decellularized matrix, natural biomaterials, and artificial synthetic materials. The biomimetic blood-brain barrier uses 3D-printed cells, including brain microvascular endothelial cells, brain microvascular pericytes, and astrocytes. The ordered layered printing of the bio-3D printing consists of the following layers from left to right: left layer of brain microvascular endothelial cells, middle layer of brain microvascular pericytes, and right layer of astrocytes. The tumor model culture area is equipped with a culture container, a container lid, a cell culture bag, a camera, and detection electrodes; The container lid is a removable screw cap; The screw cap is provided with a breathable filter membrane and an opening; The opening is for placing the cell viability detection electrode and the camera; The camera can be connected to a video recording system; The cell culture bag in the tumor model culture area can contain one or more of the following: tumor cells, tumor tissues, and tumor organoids. The tumor cells in the cell culture bag may be one or more of the following: human, animal, and cell line origins. The tumor organoids in the cell culture bag can be one or more of the following: glioma, pituitary adenoma, meningioma, and neurotumor. The cell culture bag is equipped with a filter mesh to prevent cells from passing through; The culture container is provided with interfaces on the left and right sides; The left and right interfaces are respectively connected to the system piping; The system pipeline is equipped with a drain port at the end.

[0008] Compared with the prior art, the present invention has the following beneficial effects: 1. A bio-3D printed biomimetic blood-brain barrier in vitro simulation device and its application, which realizes both the construction of a biomimetic blood-brain barrier model and the construction of an in vitro simulation device for a tumor model.

[0009] 2. The device of the present invention can be used for the evaluation of drug screening models for intracranial tumors. Liquid or drugs contained in liquid or cells contained in liquid are injected into the system pipeline through a three-way valve at the proximal end of the inlet; and then sampling and testing are performed through a three-way valve at the rear end of the biomimetic blood-brain barrier model; thus realizing the screening of drug types that can break through the biomimetic blood-brain barrier model.

[0010] 3. The device of the present invention can be used to assist in evaluating the effect of drug dosage on tumor models. Different dosages are administered into the system pipeline through a three-way valve, and the changes in the values ​​of the cell viability detection electrode are read to assist in evaluating the effect of drug dosage on tumor models.

[0011] 4. The present invention also includes a camera that can be connected to a high-definition video recording system to observe changes in the tumor model and achieve continuous visual assessment. Attached Figure Description

[0012] Figure 1 This is a schematic diagram of a bio-3D printed visualization in vitro screening model device for intracranial tumor drugs according to the present invention; wherein: 1 is the liquid inlet, 2 is the three-way valve, 3 is the biomimetic blood-brain barrier model, 4 is the tumor model culture area, 5 is the culture container, 6 is the container lid, 7 is the cell culture bag, 8 is the camera, 9 is the cell viability detection electrode, 10 is the three-way valve, and 11 is the liquid outlet. Detailed Implementation

[0013] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0014] In the description of this invention, unless otherwise specified, all reagents are commercially available and all methods used are conventional techniques in the art.

[0015] In the description of this invention, it should be understood that the terms "upper", "lower", "left", "right", "front", "rear", "inner", "bottom", "middle", "head end", "tail end", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0016] Please see Figure 1 The present invention discloses a bio-3D printed biomimetic blood-brain barrier in vitro simulation device, which is an integrated flow culture system, mainly comprising: Liquid inlet system: Equipped with inlet 1 and proximal three-way valve 2 for injecting culture medium, drugs or cell suspension; Bionic blood-brain barrier module: A bionic blood-brain barrier model 3 constructed using bio-3D printing technology, simulating the multi-layered cellular structure of the blood-brain barrier in vivo; Tumor model culture area 4: It contains a culture container 5, the container lid 6 is a detachable screw cap, equipped with a breathable filter membrane, and can accommodate a camera 8 and a cell viability detection electrode 9; The cell culture bag 7 is located inside the culture container and is used to hold tumor cells, tissues or organoids. The bag is equipped with a filter screen. Liquid discharge system: It is equipped with a remote three-way valve 10 and a liquid outlet 11 for sampling or waste liquid collection.

[0017] Example 1 A bio-3D printed visualization device for in vitro screening of intracranial tumor drugs includes an inlet 1, where the liquid is cell culture medium. A three-way valve 2 is a T-type three-way valve with a heparin cap at the external end of the valve. A biomimetic blood-brain barrier model 3 is created using bio-3D printing, with the printing material being decellularized brain tissue matrix. The cells are printed in layers from left to right: human brain microvascular endothelial cells, human brain microvascular pericytes, and human astrocytes. The tumor model culture area 4 includes a culture container 5 and a container lid 6. The lid is screw-on and has a breathable filter membrane and an opening, the size of which matches the size of the detection electrode. The cell culture bag 7 contains U87 cells. A camera 8 and a cell viability detection electrode 9 are inserted into the container lid 6, allowing for real-time readings of cell viability values. The culture container 5 has two interfaces connected to the system piping, and a three-way valve 10 serves as a sampling port, through which liquid from the system piping can be extracted. The cell culture medium entering from the inlet 1 flows through the entire system pipeline to the end, and finally the liquid flows out from the outlet 11, which can be connected to a waste liquid collection device.

[0018] In this scheme, different doses of temozolomide drug solution can be injected into the three-way valve 2, and the data changes of the cell viability detection electrode and the real-time recording of the high-definition camera system can be used to help interpret and screen the drug dose that meets the best treatment effect.

[0019] Example 2 See Figure 1 The effect evaluation of the test drug on the glioma tumor model disclosed in this embodiment is a variation of Example 1. The difference is that the test drug is injected into the three-way valve 2. The data changes of the cell viability detection electrode and the real-time recording of the high-definition camera system are used to help interpret the effect of the test drug on the glioma tumor model.

[0020] Example 3 See Figure 1 The effect evaluation of CD8+ T lymphocytes on the glioma tumor model disclosed in this embodiment is a variation of Example 1. The difference is that CD8+ T lymphocyte solution is injected into the three-way valve 2. The data changes of cell viability detection electrode and the real-time recording of high-definition camera system are used to help interpret the penetration of CD8+ T lymphocytes across the biomimetic blood-brain barrier model, and thus evaluate the effect of CD8+ T lymphocytes on the glioma tumor model.

[0021] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A bio-3D printed visualization in vitro screening model device for intracranial tumor drugs, characterized in that, It includes a liquid inlet module, a biomimetic blood-brain barrier module, a tumor culture module, and a liquid outlet module connected in sequence; The biomimetic blood-brain barrier module is constructed using bio-3D printing technology. The tumor culture module includes a culture container, a cell culture bag inside it, and a camera and cell viability detection electrode mounted on the container lid.

2. The bio-3D printed visualization in vitro screening model device for intracranial tumor drugs according to claim 1, characterized in that: The cell culture bag is equipped with a filter mesh.

3. The bio-3D printed visualization in vitro screening model device for intracranial tumor drugs according to claim 1, characterized in that: The biomimetic blood-brain barrier module is composed of a layer of cerebral microvascular endothelial cells, a layer of pericytes, and a layer of astrocytes printed sequentially.

4. The bio-3D printed visualized in vitro screening model device for intracranial tumor drugs according to claim 1, characterized in that: Both the inlet and outlet modules are equipped with three-way valves, and the external ports of the three-way valves are equipped with heparin caps.

5. The application of a bio-3D printed visualization in vitro screening model device for intracranial tumor drugs as described in any one of claims 1-4 in the screening of intracranial tumor drugs.

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