CN-121994885-A - Hepatic fibrosis organ chip sensing device for pathology assessment and drug screening

CN121994885ACN 121994885 ACN121994885 ACN 121994885ACN-121994885-A

Abstract

The invention discloses a hepatic fibrosis organ chip sensing device for pathological evaluation and drug screening, which belongs to the technical field of microfluidics and comprises a top cover, an upper culture layer, a porous membrane, a lower culture layer and a glass substrate electrochemical electrode. The sensing device combines a hepatic fibrosis organ chip and a three-electrode detection system, and detects gel permeability change based on electrochemical cyclic voltammetry to indirectly evaluate the hepatic fibrosis progress so as to perform in-vitro rapid screening of hepatic fibrosis potential drugs. The hepatic fibrosis organ chip sensing device provided by the invention provides a novel and efficient research means for real-time monitoring and quantitative evaluation of an in-vitro hepatic fibrosis model. Solves the problems that in the prior art, the dynamic and trans-scale substance transmission characteristics in a three-dimensional tissue model are accurately quantized, so that the micro-environment change led by extracellular matrix reconstruction is difficult to reflect in situ and the hepatic fibrosis progress is difficult to detect.

Inventors

WANG YANYAN
HU XINYANG
Jiang Kaishi

Assignees

天津大学

Dates

Publication Date: 20260508
Application Date: 20260129

Claims (8)

1. The liver fibrosis organ chip sensing device for pathology assessment and drug screening is characterized by comprising an organ chip and an electrochemical detection module, wherein the organ chip sequentially comprises a top cover (1), an upper culture layer (2), a porous membrane (3), a lower culture layer (4) and a glass substrate electrochemical electrode (5) from top to bottom, the electrochemical detection module comprises a traditional single-channel electrochemical workstation, the top cover and the upper culture layer are provided with liquid through holes (6), the upper culture layer and the lower culture layer are provided with liquid through channels (7), and the glass substrate electrochemical electrode is connected with an electrode chuck corresponding to the electrochemical workstation.
2. The use of a hepatic fibrosis organ chip sensing device for pathological evaluation and drug screening, which is characterized in that the hepatic fibrosis organ chip sensing device for pathological evaluation and drug screening is adopted, and is characterized in that the sensing device is combined with a hepatic fibrosis organ chip and a three-electrode detection system, gel permeability change is detected based on electrochemical cyclic voltammetry, and the hepatic fibrosis progress is indirectly evaluated, so that in-vitro rapid screening of hepatic fibrosis potential drugs is performed.
3. The use of a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening of claim 2, wherein the electrolyte detected by electrochemical cyclic voltammetry is a gel gelled from a prepolymer of cells and collagen I solution.
4. Use of a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening of claim 2, wherein the assessment comprises the steps of: s1, establishing a concentration-current response relation through a cyclic voltammetry curve, and calibrating a detection system, S2, evaluating the liver fibrosis progress according to the concentration-current response relation by measuring the concentration of electroactive substances; S3, determining cyclic voltammograms under different permeation duration, and evaluating the permeation rate by fitting to evaluate the liver fibrosis progress.
5. The use of a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening of claim 4, wherein step S1 comprises the steps of: And (3) adding electroactive substances with different concentrations into the culture layer under the liver organ chip, measuring electrochemical response of the culture layer by using a cyclic voltammetry to obtain cyclic voltammetry curves corresponding to different concentrations, extracting oxidation peak current, and establishing a concentration-current response relation to obtain a linear fitting equation.
6. The use of a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening of claim 4, wherein step S2 comprises the steps of: the concentration of electroactive material measured by electrochemistry reflects the permeability change, the electroactive material is added to the organ-on-chip culture layer with or without liver fibrosis, the concentration of electroactive material permeated from the upper culture layer into the lower culture layer is measured, and the permeability change is estimated by using the oxidation peak current according to the concentration-current response relationship, so as to estimate the liver fibrosis progress.
7. The use of a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening of claim 4, wherein step S3 comprises the steps of: further introducing the permeation rate, measuring cyclic voltammograms under different permeation time lengths, establishing a fitting relation between oxidation peak current and permeation time, evaluating the permeation rate by using the slope of a fitting straight line, and evaluating the liver fibrosis progress by combining permeation change.
8. The use of a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening of claim 2, wherein the drug screening comprises the steps of adding different drug stimuli to the hepatic fibrosis organ chip, continuously culturing the same in an upper culture layer, comparing the presence or absence of the drug stimulus group, assessing the effect of the applied drug on the hepatic fibrosis pathology model, and performing in vitro rapid screening of the hepatic fibrosis potential drug by assessing the change in permeability after drug stimulation.

Description

Hepatic fibrosis organ chip sensing device for pathology assessment and drug screening Technical Field The invention relates to the technical field of microfluidics, in particular to a hepatic fibrosis organ chip sensing device for pathological evaluation and drug screening. Background Liver fibrosis is a pathological change process of extracellular matrix (Extracellular matrix, ECM) of liver caused by chronic liver injury and other factors, and is mainly represented by excessive generation and scarring of ECM (collagen, fiber and the like) after hepatic stellate cell activation, and finally, the liver fibrosis is developed into irreversible end-stage diseases such as liver cirrhosis and the like. At present, the specific pathogenesis of liver fibrosis is not clear, the liver fibrosis process is not easy to monitor, and an effective direct anti-fibrosis drug is also lacking. The Organ-on-a-Chip (OoC) technology can combine the micro-fluidic technology with tissue engineering, simulate the local spatial structure of micro-units of a microscale Organ through structural design, provide physiological simulated shear flow to promote cell population distribution and functional expression improvement, and provide a highly bionic in vitro model for drug and disease research. In the analysis process of the organ chip model, the traditional methods, such as immunofluorescence staining, western blotting, enzyme-linked immunosorbent assay and the like, often require the processes of fixing, preprocessing and dye incubation of biological samples, have complicated steps and long time consumption, and are difficult to continuously monitor in-situ and dynamic changes in the experimental process. Electrochemical analysis technology is widely used in cell biology research for the detection of lactic acid, glucose and various enzyme metabolites due to its rapid response and real-time monitoring capabilities. Methods of combining cell electrical impedance sensing, transepithelial/endothelial resistance measurement and the like in organ chips have been studied to achieve permeability determination of cell barriers. However, the above-mentioned conventional electrochemical method still has limitations in precisely quantifying dynamic, trans-scale substance transmission characteristics in a three-dimensional tissue model, and is difficult to reflect the micro-environmental changes dominated by extracellular matrix reconstruction in a noninvasive and in situ manner, and hepatic fibrosis progress is not easy to detect. Disclosure of Invention Therefore, the invention provides a hepatic fibrosis organ chip sensing device for pathological evaluation and drug screening, which is used for solving the problems that in the prior art, the micro-environment change dominated by extracellular matrix reconstruction is difficult to reflect in situ and the hepatic fibrosis progress is difficult to detect because of the limitation on the aspect of precisely quantifying dynamic and trans-scale substance transmission characteristics in a three-dimensional tissue model. In order to achieve the above object, the present invention provides the following technical solutions: According to a first aspect of the invention, a hepatic fibrosis organ chip sensing device for pathology assessment and drug screening is provided, the organ chip sensing device comprises an organ chip and an electrochemical detection module, the organ chip sequentially comprises a top cover 1, an upper culture layer 2, a porous membrane 3, a lower culture layer 4 and a glass substrate electrochemical electrode 5 from top to bottom, the electrochemical detection module comprises a traditional single-channel electrochemical workstation, the top cover and the upper culture layer are provided with liquid through holes 6, the upper culture layer and the lower culture layer are provided with liquid through channels 7, and the glass substrate electrochemical electrode is connected with a corresponding electrode chuck of the electrochemical workstation. According to a second aspect of the present invention, there is provided the use of a liver fibrosis organ chip sensing device for pathology assessment and drug screening, the liver fibrosis organ chip sensing device for pathology assessment and drug screening being employed in combination with a liver fibrosis organ chip and three-electrode detection system, the liver fibrosis progress being indirectly assessed based on electrochemical cyclic voltammetry detection of gel permeability changes, for in vitro rapid screening of potential drugs for liver fibrosis. Further, the electrolyte detected by the electrochemical cyclic voltammetry is gel obtained by gelation of a prepolymer of cells and a collagen I solution. Further, the evaluation includes the steps of: s1, establishing a concentration-current response relation through a cyclic voltammetry curve, and calibrating a detection system; s2, evaluating the liver fibrosis progress accordin