Evaluation method for testing toxic and side effects of medicine through biological organ model

Abstract

The invention relates to the technical field of drug toxicity evaluation, in particular to an evaluation method for examining toxic and side effects of drugs through a biological organ model. The method comprises the following steps: pretreating a cell line, and carrying out mixed culture with a skeleton structure; connecting an organ model through a biochip; designing a micro-channel network to form a circulation simulation system, and adjusting micro-environment parameters; embedding a biosensor into the biological organ model, and collecting data; adding drugs with different concentrations, and evaluating according to data. According to the method, firstly, the liver, the kidney and the heart are constructed, then the three organ models are connected through the microfluidic technology, microenvironment factors such as micro blood vessels and lymphatic vessels are introduced into the connecting positions, finally, the biosensor technology is introduced, the cell state, metabolites and microenvironment parameters in all the organ models are monitored in real time, and the three organ models are obtained. Through the multi-dimensional biological organ model, the influence of the medicine on different organs can be evaluated more comprehensively.

Description

Technical Field

[0001] The present invention relates to the technical field of drug toxicity evaluation, and in particular to a method for evaluating drug toxicity and side effects by using a biological organ model. Background Art

[0002] Drugs refer to substances that can change or identify the physiological functions and pathological states of the body and can be used to prevent, treat, diagnose diseases, or purposefully regulate the physiological functions of the body. Poisons refer to substances that can cause damage to the animal body. However, "all drugs are poisonous". When drugs are used in excess of a certain dose or are used improperly, they will have toxic effects. Therefore, there is no absolute boundary between drugs and poisons. The only difference between the two is the difference in dosage. Long-term use or excessive dosage of drugs may become poisons.

[0003] For example, CN117126914A involves a method for evaluating drug toxicity in a liver organoid model that reflects toxic and side effects, including: separation and culture of mouse liver cells; construction of a three-dimensional bioprinted mouse liver framework; addition of other mouse liver cell types; simulation of mouse liver blood supply; simulation of mouse liver metabolic enzyme system; drug treatment and toxicity evaluation; data collection and analysis. This invention only constructs liver organoids through cells, and evaluates the toxic and side effects of drugs through the reaction of liver organoids when taking drugs. However, in addition to liver organoids, animals also have other organs such as the heart and kidneys. Multiple organs in animals complement each other, and it is impossible for drugs to only affect liver organoids when used. Therefore, the above-mentioned drug toxicity evaluation method is not comprehensive enough, and thus misjudges the clinical use effect of drugs.

[0004] Therefore, in order to make a more comprehensive evaluation of the toxic and side effects of drugs, a method for evaluating the toxic and side effects of drugs using a biological organ model is proposed. Summary of the invention

[0005] The purpose of the present invention is to provide a method for evaluating the toxic and side effects of drugs by using a biological organ model to solve the problems raised in the above-mentioned background technology.

[0006] To achieve the above object, the present invention provides a method for evaluating the toxic and side effects of a drug by testing a biological organ model, comprising the following steps:

[0007] S1. Pre-treating multiple cell lines, and then planting and culturing different cell lines with skeleton structures to form a three-dimensional biological organ model;

[0008] S2, connect different organ models through biochips for integration, and then introduce micro blood vessels and lymphatic vessels to connect different organ models;

[0009] S3, design a microchannel network to connect micro blood vessels and lymphatic vessels, cooperate with the biochip to form a circulation simulation system, and set a microvalve controller to adjust the microenvironment parameters;

[0010] S4, embed biosensors into biological organ models to contact cells and microenvironment, and establish a real-time monitoring system to collect data;

[0011] S5. Add different concentrations of drugs to the biological organ model through the controlled variable method, and evaluate the drug toxicity and side effects based on the drug response data.

[0012] As a further improvement of the present technical solution, in S1, cell pretreatment includes cell expansion, differentiation and function enhancement, and biodegradable scaffold materials or bioprinting technology are used to construct the skeleton structure of the organ, and then hepatocyte cell lines, renal tubular cell lines and cardiomyocyte cell lines are selected to culture to form independent liver, kidney and heart.

[0013] As a further improvement of the technical solution, in S2, a biochip with multiple channels is used when integrating different organ models, and the different organ models are loaded into different channels of the biochip respectively.

[0014] As a further improvement of the technical solution, in S2, micro blood vessels and lymphatic vessels are introduced into the biological organ model through a micro bioreactor, and the micro blood vessels and lymphatic vessels are in direct contact with cells of the organ model.

[0015] As a further improvement of the present technical solution, in S2, the materials of the micro blood vessels and lymphatic vessels are polymers or biodegradable materials.

[0016] As a further improvement of the technical solution, in S3, the diameter of the microchannel network matches the diameter of the microblood vessels and lymphatic vessels.

[0017] As a further improvement of the present technical solution, in S3, the circulation simulation system and the fluid flow speed and flow direction in the channel are controlled by a microvalve controller to adjust the microenvironment parameters, and the microenvironment parameters include fluid flow rate, oxygen concentration and pH value.

[0018] As a further improvement of the present technical solution, in S4, the biosensor includes a biochemical sensor, a bioimaging sensor, and a bioelectric sensor, and the collected data includes cell status, metabolite concentration, and microenvironment parameters.

[0019] As a further improvement of the technical solution, in S5, the microenvironment parameters of the biological organ model and the circulation simulation system are first adjusted, and under the current microenvironment parameter conditions, drugs of different concentrations are added to the biological organ model.

[0020] As a further improvement of the present technical solution, in S5, the drug reaction data includes drug transmission rate, metabolite concentration change value and microenvironment parameter change value.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] In this evaluation method for testing drug toxicity and side effects through biological organ models, the liver, kidneys and heart are first constructed, and then the three organ models are connected using microfluidics technology, and microenvironmental factors such as microvessels and lymphatic vessels are introduced at the connection. Finally, biosensor technology is introduced to monitor the cell status, metabolites and microenvironmental parameters in each organ model in real time. Through such a multi-dimensional biological organ model, the effects of drugs on different organs can be more comprehensively evaluated. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a flow chart of the present invention. DETAILED DESCRIPTION

[0024] The following will be combined with the accompanying drawings in the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] The existing technical solutions only construct liver organoids through cells and evaluate the toxic and side effects of drugs through the reactions of liver organoids when taking drugs. However, in addition to liver organoids, animals also have other organs such as heart and kidneys. Multiple organs in animals complement each other. It is impossible for drugs to only affect liver organoids when used. Therefore, the above-mentioned drug toxicity evaluation method is not comprehensive enough.

[0026] Therefore, please refer to Figure 1 As shown, the purpose of this embodiment is to provide an evaluation method for testing drug toxicity and side effects through a biological organ model, comprising the following steps:

[0027] S1. First, select appropriate cell types, including cells that mainly constitute the organ and supporting cells, to ensure representativeness and functionality; pre-treat the cells, including cell expansion, differentiation and function enhancement, to improve their stability and activity in the three-dimensional biological organ model; prepare biological scaffolds or biological inks as a matrix for cell growth and tissue formation; biological scaffolds can be natural or synthetic scaffold materials, such as biodegradable polymers or natural collagen; biological inks can be biological materials containing cell support factors and growth factors; mix or plant the pre-treated cells with the biological scaffold or biological ink to ensure that the cells are evenly distributed and in full contact with the scaffold or ink; assemble cells layer by layer according to the anatomical structure and cell histological characteristics of the organ to simulate the real organ structure; place the assembled biological organ model in an in vitro culture environment to provide suitable growth conditions and environmental parameters, such as temperature, humidity and gas composition; during the culture process, adjust the culture medium components, growth factors and cytokines as needed to promote cell growth and tissue formation;

[0028] In this embodiment, a biodegradable scaffold material or bioprinting technology is used to construct the skeleton structure of the organ, and a liver cell line, a renal tubular cell line and a cardiomyocyte cell line are selected, and these cells are pretreated to enhance their metabolic and anatomical characteristics. Then, a biodegradable polymer scaffold is prepared, and different cell lines are mixed with the scaffold for implantation. During the implantation process, different cell lines are assembled in layers according to the anatomical structure of the organ to simulate the real organ tissue structure, and finally an independent liver, kidney and heart are formed. The assembled biological organ model is placed in a culture medium for in vitro culture, and the composition and growth factors of the culture medium are adjusted to promote the growth and metabolic activity of the organ cells.

[0029] S2. A biochip with multiple channels is used. In this embodiment, a biochip with three channels is used to accommodate liver, kidney and heart models respectively. Then, the pre-cultured liver cell line, kidney cell line and heart cell line are loaded into different channels of the biochip respectively. The liquid flow between the channels is regulated by microfluidic technology to ensure the material transfer and interaction between the organ models, and microenvironmental factors such as micro blood vessels and lymphatic vessels at the micrometer scale are introduced into the biochip. The micro blood vessels and lymphatic vessels can use microfluidic chip manufacturing technology or 3D printing technology, and select suitable materials, such as polymers or biodegradable materials, to ensure that the model has good biocompatibility; the designed micro blood vessel and lymphatic vessel models are embedded in the multi-dimensional biological organ model and connected to the main organ model to ensure that the micro blood vessels and lymphatic vessels are in direct contact with the cells of the organ model to achieve material exchange and signal transmission; the microfluidic system is then used to control the fluid flow in the micro blood vessels and lymphatic vessels to simulate the blood and lymph circulation process in the body;

[0030] S3. Use microfluidic technology to design a microchannel network that matches the diameter of microvessels and lymphatic vessels, connect the microvessels and lymphatic vessels, and cooperate with biochips to form a circulation simulation system. Then use a microvalve controller to adjust the circulation simulation system and the fluid flow in the channel to simulate the flow direction and speed of blood and lymph in the body; by adjusting the flow speed and direction, changing the physical and chemical parameters of the microenvironment such as flow rate, oxygen concentration and pH value to simulate the microenvironment conditions under different physiological states, it can more realistically simulate the interaction between human organs and provide a more reliable experimental platform for drug development and toxicity assessment;

[0031] S4. Select biosensors suitable for monitoring multidimensional biological organ models, such as biochemical sensors, bioimaging sensors, bioelectric sensors, etc., to ensure that the biosensors have high sensitivity, high selectivity and good stability to meet the needs of real-time monitoring; embed the selected biosensors into appropriate locations of the multidimensional biological organ model; ensure that the biosensors can effectively contact the cells and microenvironment in the model to obtain accurate monitoring data; establish a real-time monitoring system, including sensor connection and signal acquisition system, to achieve real-time acquisition and recording of biosensor output signals; ensure the stability and high responsiveness of the monitoring system, and enable real-time monitoring and data recording; conduct real-time analysis and interpretation of the collected monitoring data, including cell status, metabolite concentrations and microenvironment parameters, and use statistical and machine learning methods to identify the response characteristics and change patterns of the biological organ model to provide support for subsequent experimental design and data interpretation;

[0032] S5. Test the toxic and side effects of drugs by controlling variables. First, in order to test the effect of drug concentration on drug toxic and side effects, adjust the physical and chemical parameters of the microenvironment such as fluid flow rate, oxygen concentration and pH value so that the biological organ model and the circulation simulation system can simulate the human microenvironment. Then add drugs of different concentrations to the biological organ model. After the addition of drugs, the changes in the cell states of different organ models, microvessels and lymphatic vessels in the biological organ model are detected by biosensors. The drug transmission rate in the biological organ model, as well as the concentration changes of cell metabolites and the changes in the microenvironment under drug stimulation are measured. Secondly, due to the physical differences between people in different states and between people, there are different physiological states. In order to examine the differences in drug toxicity and side effects under different physiological states, the physical and chemical parameters of the microenvironment such as fluid flow rate, oxygen concentration and pH value are fine-tuned, and then different concentrations of drugs are added. The above process is repeated to monitor the changes in the oxygen concentration and metabolite concentration of cells in real time, understand the metabolic state and microenvironment parameters of the multi-dimensional biological organ model, and perform real-time analysis and interpretation based on the monitored data to identify the changing patterns of cell states and microenvironment parameters, thereby achieving a comprehensive evaluation of drug toxicity and side effects.

[0033] To summarize, first, appropriate cell lines or tissue engineering models are selected. Then, biodegradable scaffold materials or bioprinting technology are used to construct three-dimensional structures of the liver, kidneys, and heart. Next, microfluidics technology is used to connect the three organ models, and microenvironmental factors such as microvessels and lymphatic vessels are introduced at the connection. Finally, biosensor technology is introduced to monitor the cell status, metabolites, and microenvironmental parameters in each organ model in real time. Through such a multidimensional biological organ model, the effects of drugs on different organs can be more comprehensively evaluated, providing more reliable guidance for drug development.

[0034] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and descriptions are only preferred examples of the present invention and are not intended to limit the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A method for evaluating drug toxicity and side effects by testing a biological organ model, characterized in that: The following steps are involved: S1. Pre-treating multiple cell lines, and then planting and culturing different cell lines with skeleton structures to form a three-dimensional biological organ model; S2, connect different organ models through biochips for integration, and then introduce micro blood vessels and lymphatic vessels to connect different organ models; S3, design a microchannel network to connect micro blood vessels and lymphatic vessels, cooperate with the biochip to form a circulation simulation system, and set a microvalve controller to adjust the microenvironment parameters; S4, embed biosensors into biological organ models to contact cells and microenvironment, and establish a real-time monitoring system to collect data; S5. Add different concentrations of drugs to the biological organ model through the controlled variable method, and evaluate the drug toxicity and side effects based on the drug response data.

2. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S1, cell pretreatment includes cell expansion, differentiation and function enhancement, and biodegradable scaffold materials or bioprinting technology are used to construct the skeleton structure of the organ, and then hepatocyte cell lines, renal tubular cell lines and cardiomyocyte cell lines are selected to culture to form independent liver, kidney and heart.

3. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S2, a biochip with multiple channels is used to integrate different organ models, and different organ models are loaded into different channels of the biochip respectively.

4. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S2, micro blood vessels and lymphatic vessels are introduced into the biological organ model through a micro bioreactor, and the micro blood vessels and lymphatic vessels are in direct contact with cells of the organ model.

5. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S2, the materials of the micro blood vessels and lymphatic vessels are polymers or biodegradable materials.

6. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S3, the diameter of the microchannel network matches the diameter of the microblood vessels and lymphatic vessels.

7. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S3, the circulation simulation system and the fluid flow speed and flow direction in the channel are controlled by a microvalve controller to adjust the microenvironment parameters, and the microenvironment parameters include fluid flow rate, oxygen concentration and pH value.

8. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S4, the biosensor includes a biochemical sensor, a bioimaging sensor, and a bioelectric sensor, and the collected data includes cell status, metabolite concentration, and microenvironment parameters.

9. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S5, the microenvironment parameters of the biological organ model and the circulation simulation system are adjusted first, and under the current microenvironment parameter conditions, drugs of different concentrations are added to the biological organ model.

10. The method for evaluating drug toxicity and side effects by testing a biological organ model according to claim 1, characterized in that: In S5, the drug reaction data includes drug transmission rate, metabolite concentration change value and microenvironment parameter change value.

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