CN111458491A - An organ drug test chip - Google Patents

Abstract

The invention discloses an organ drug testing chip, which relates to the technical field of organ chips. The invention includes an upper-layer chip and a lower-layer chip; the bottom surface of the upper-layer chip is provided with at least three recessed Tesla micro-mixers, a first inlet channel and a first The main channel, the Tesla micro-mixers are arranged in parallel, the channel inlets of the Tesla micro-mixers are all connected with the first main channel, the first inlet channels are all located in the middle of the adjacent Tesla micro-mixers, and the first inlet channel One end is connected with the first main channel; the upper surface of the lower chip is provided with a recessed cell culture chamber and a first outlet channel, the first outlet channel is communicated with the cell culture chamber, and the channel outlet of the Tesla micromixer is connected with the cell culture chamber coincide. The beneficial effects of the present invention are: the organ drug test chip provided by the present invention can automatically form multiple drug concentration gradients, so as to directly study the organ toxicity of the drug under different concentration gradients, or the organ toxicity of multiple mixed drugs under different concentrations.

Description

An organ drug test chip

technical field

The invention relates to the technical field of organ chips, in particular to an organ drug testing chip.

Background technique

A key factor in early stage drug development or drug candidate screening is the identification of its toxic side effects, which are the main reason for a drug to withdraw from clinical trials or the market. According to statistics, the successful development of a new type of drug generally requires investment of 3-5 billion US dollars in research and development funds, which lasts 12-15 years, and the vast majority (about 90%) of drugs will fail miserably in the clinical trial stage. One-third of drugs that fail are toxic to human organs. Currently, drug development is mainly based on animal models. Due to species differences and differences in drug metabolism pathways, animal models cannot well predict drug effects in humans. For example, cytochrome P450 is essential for most exogenous detoxification in the liver, metabolism Enzyme induction, inhibition mechanisms and their clearance and excretion rates vary from species to species, making it difficult to extrapolate relevant parameters from animals. In addition, genetic diversity also challenges the prediction of drug metabolism. Accordingly, there is an urgent need for more reliable drug testing models that are representative of human conditions.

Although the 2D monolayer cell culture model provides an important contribution to drug research, this model is too simple to represent the complex tissue interactions in vivo. In recent years, the development of microfabrication and microfluidics has provided options for designing small and precise systems suitable for cell culture applications, which can provide microenvironments similar to those in the human body (three-dimensional tissue culture environment, nutrient supply, waste products). timely clearance and appropriate shear rate) to simulate human organs in vitro, thus bridging the gap between preclinical and 2D cell culture.

Compared with single-organ-specific functions, novel microfluidic-based platforms are more widely used to characterize interactions between multiple organs, such as liver-gut, liver-lung, and liver-kidney models. Among them, liver toxicity and nephrotoxicity are the two main reasons that lead to the withdrawal of drugs from the market. Patent CN110628566A discloses an organ chip, but the existing organ chip cannot test drugs with different concentration gradients.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is to provide an organ drug test chip capable of testing drugs of different concentrations.

The present invention realizes and solves the above-mentioned technical problems through the following technical means:

An organ drug testing chip, comprising an upper-layer chip and a lower-layer chip arranged in sequence from top to bottom;

The bottom surface of the upper chip is provided with at least three recessed Tesla micro-mixers, a first inlet channel and a first main channel, the Tesla micro-mixers are arranged in parallel, and the Tesla micro-mixers are arranged in parallel. The channel inlets are all connected with the first main channel, the first inlet channels are all located in the middle of the adjacent Tesla micromixer, and one end of the first inlet channel is communicated with the first main channel;

The upper surface of the lower chip is provided with a recessed cell culture chamber and a first outlet channel, the first outlet channel communicates with the cell culture chamber, and the channel outlet of the Tesla micromixer coincides with the cell culture chamber.

Working principle: put the prepared organ tissue into the cell culture chamber, install the upper chip on the lower chip, different concentrations of drugs enter the first main channel from different first inlet channels, due to the recessed Tesla micromixer They are arranged in parallel, and the first inlet channels are located in the middle of the adjacent Tesla micromixers. After mixing, the drugs in the adjacent first inlet channels can simultaneously enter the Tesla between the adjacent first inlet channels. In the recessed channel of the micro-mixer, when the number of Tesla micro-mixers is two, the Tesla micro-mixer contains only one concentration of the drug, and when the number of Tesla micro-mixers is at least two At the time of three, different concentrations of drugs flow into the Tesla micromixer on both sides at the same time, and are quickly and efficiently mixed by the Tesla micromixer to form a stable concentration gradient drug. The channel outlet coincides with the cell culture chamber, and the drug can enter the cell culture chamber from the channel outlets of the plurality of Tesla micromixers, and finally flow into the first outlet channel communicated with the cell culture chamber. Beneficial effect: The organ drug test chip provided by the present invention can automatically form multiple drug concentration gradients through the Tesla micro-mixer recessed on the bottom surface of the upper chip, so as to directly study the organ damage under different concentration gradients of the drug, or a variety of mixed drugs Organ damage at different concentrations.

The mixing channel of the existing drug test chip is a simple mixing channel, and the mixing efficiency is not high, and it is difficult to form a stable drug concentration gradient. The stable drug concentration gradient improves the efficiency and reliability of drug testing.

The invention directly etches the concave channel on the organ drug chip, which can reduce the volume of the organ drug chip.

Preferably, the Tesla micro-mixer includes a multi-stage Tesla micro-mixer, the multi-stage Tesla micro-mixers are sequentially connected in series, and the outlet of any Tesla micro-mixer of the previous stage The channel is located in the middle of the inlet channel of the adjacent Tesla micromixer of the latter stage, and the number of the Tesla micromixer of the latter stage is one more than the number of the Tesla micromixer of the previous stage.

Preferably, the multi-stage Tesla micro-mixer includes a first-stage Tesla micro-mixer and a second-stage Tesla micro-mixer; the channel inlets of the first-stage Tesla micro-mixer are The first main channel is connected, and the first inlet channel is located in the middle of the adjacent first-stage Tesla micromixer;

The channel inlet of the second-stage Tesla micro-mixer is provided with a second main channel, the channel inlets of the second-stage Tesla micro-mixer are all communicated with the second main channel, and the first-stage Tesla micro-mixer is connected with the second main channel. The channel outlet of the pull micro-mixer is located in the middle of the adjacent second-stage Tesla micro-mixer, and the channel outlet of the second-stage Tesla micro-mixer coincides with the cell culture chamber.

Preferably, the number of the first-stage Tesla micro-mixers is three, the number of the second-stage Tesla micro-mixers is four, and the number of the cell culture chambers is four .

Preferably, the cell culture chamber includes a first cell culture chamber and a second cell culture chamber, the first cell culture chamber communicates with the second cell culture chamber, and the channel outlet of the Tesla micromixer is connected to the first cell culture chamber. The cell culture chambers overlap, and the second cell culture chamber communicates with one end of the first outlet channel.

Working principle: Put the prepared liver and kidney tissues into the first cell culture room and the second cell culture room, respectively, to study the liver toxicity and liver-related nephrotoxicity of drugs under different concentration gradients, or mix drugs with different concentrations Hepatotoxicity and its liver-related nephrotoxicity.

Beneficial effects: The chip in the present invention can form a variety of drug concentration gradients, and contains both hepatocyte chambers and kidney cell chambers, so that it can directly study the liver toxicity and liver-related nephrotoxicity under different drug concentrations, so that the Improve the efficiency and reliability of drug testing.

Preferably, the number of the first cell culture chambers is the same as the number of the second-stage Tesla micromixers, and the number of the second cell culture chambers is the same as the number of the first cell culture chambers.

Preferably, the other end of the first outlet channel communicates with the outer wall of the lower chip.

Beneficial effects: the cell metabolites can be collected through the first outlet channel, and the cell state can be monitored in real time through the cell metabolites.

Preferably, a first channel is provided on one side of the upper-layer chip, the cross-section of the first channel is L-shaped, one end of the first channel passes through the sidewall of the upper-layer chip, and the other end of the first channel The other end of the first channel communicates with the first inlet channel through the lower surface of the upper chip.

Preferably, the channel depth of the Tesla micromixer is smaller than the thickness of the upper chip.

Preferably, the depth of the cell culture chamber and the depth of the first outlet channel are smaller than the thickness of the underlying chip.

Preferably, the upper-layer chip is fixed to the lower-layer chip by screws.

The working principle of the present invention: put the prepared organ tissue into the cell culture chamber, install the upper chip on the lower chip, and the drugs of different concentrations enter the first main channel from different first inlet channels, and the adjacent first inlet channels After being mixed, the drug in the inside enters the Tesla micro-mixer between the adjacent first inlet channels, and is quickly and efficiently mixed by the Tesla micro-mixer to form a stable gradient concentration of the drug, and then passes through multiple Tesla micro-mixers. Pull the channel outlet of the micromixer into the cell culture chamber and finally flow out from the first outlet channel.

The advantage of the present invention is that the organ drug test chip provided by the present invention can automatically form multiple drug concentration gradients, so as to directly study the organ toxicity under different concentration gradients of the drug, or the organ damage under different concentrations of multiple mixed drugs.

The mixing channel of the existing drug test chip is a simple mixing channel, and the mixing efficiency is not high, and it is difficult to form a stable drug concentration gradient. The stable drug concentration gradient improves the efficiency and reliability of drug testing.

Description of drawings

1 is an exploded view of an organ drug test chip in Embodiment 1 of the present invention;

2 is an exploded perspective view of the organ drug test chip in Embodiment 1 of the present invention;

3 is a schematic structural diagram of an upper-layer chip in Embodiment 1 of the present invention;

4 is a bottom view of an upper-layer chip in Embodiment 1 of the present invention;

5 is a schematic structural diagram of a lower-layer chip in Embodiment 1 of the present invention;

6 is a top view of a lower-layer chip in Embodiment 1 of the present invention;

FIG. 7 is the survival rate of hepatocytes in the first cell culture chamber in Example 2 of the present invention;

In the figure: upper chip 11; first channel 12; first inlet channel 13; first main channel 14; first stage Tesla micromixer 15; second main channel 16; second stage Tesla micromixer 17; second channel 18; lower chip 21; first cell culture chamber 22; second cell culture chamber 23; first outlet channel 24.

Detailed ways

In order to make the purposes, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are part of the present invention. examples, but not all examples. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

It should be noted that when an element is referred to as being "fixed to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present.

It should be noted that, in this document, if there are relational terms such as first and second, etc., it is only used to distinguish one entity or operation from another entity or operation, and does not necessarily require or imply these entities or operations There is no such actual relationship or order between them. Moreover, the terms "comprising", "comprising" or any other variation thereof are intended to encompass a non-exclusive inclusion such that a process, method, article or device that includes a list of elements includes not only those elements, but also includes not explicitly listed or other elements inherent to such a process, method, article or apparatus. Without further limitation, an element qualified by the phrase "comprising a..." does not preclude the presence of additional identical elements in a process, method, article or apparatus that includes the element.

Example 1

An organ drug testing chip, as shown in Figures 1 and 2, includes an upper layer chip 11 and a lower layer chip 21 arranged in sequence from top to bottom. The upper surface of 21 is provided with corresponding threaded holes, and the upper chip 11 and the lower chip 21 are connected by screws.

As shown in FIG. 3 and FIG. 4 , the bottom surface of the upper chip 11 is provided with an upwardly recessed Tesla micromixer, an upwardly recessed first inlet channel 13 and an upwardly recessed first main channel 14. The Tesla micromixer It is composed of Tesla (Tesla) structure. The Tesla micro-mixer is the existing technology. The Coanda effect is used to make the fluid flow back, which can greatly enhance the chaotic convection and improve the mixing effect. The depths of the first inlet channel 13 and the upwardly recessed first main channel 14 are both smaller than the thickness of the upper chip 11 .

In order to form more substance concentration gradients, the Tesla micro-mixer is composed of multiple stages, and the multi-stage Tesla micro-mixers are connected in series in sequence, and the outlet channel of any Tesla micro-mixer of the previous stage is located in the rear. In the middle of the inlet channel of the adjacent Tesla micro-mixers of the first stage, the number of Tesla micro-mixers of the subsequent stage is one more than the number of Tesla micro-mixers of the previous stage. The mixers are the same size.

In this embodiment, the Tesla micro-mixer is composed of two stages, including a first-stage Tesla micro-mixer 15 and a second-stage Tesla micro-mixer 17. In this embodiment, the first-stage Tesla micro-mixer The number of the mixers 15 is three, the number of the second-stage Tesla micro-mixers 17 is four, and the number of the first inlet channels 13 is two.

The channel inlets of the three first-stage Tesla micro-mixers 15 are all communicated with the side walls of the first main channel 14, wherein the channel inlets of the two first-stage Tesla micro-mixers 15 are respectively connected with the first main channel 14. The two ends of the side wall are connected, the channel inlet of the other first-stage Tesla micro-mixer 15 is located in the middle of the side wall of the first main channel 14, and the first inlet channels 13 are respectively located in the adjacent first-stage Tesla micro-mixers. In this embodiment, the axis of the first-stage Tesla micromixer 15 is perpendicular to the axis of the first main channel 14 .

The channel inlets of the four second-stage Tesla micro-mixers 17 are all communicated with the side walls of the second main channel 16, and the channel inlets of the four second-stage Tesla micro-mixers 17 are evenly distributed in the second main channel 16. , wherein the channel inlets of the two second-stage Tesla micromixers 17 are respectively communicated with the side walls of the second main channel 16, and the channel outlets of the three first-stage Tesla micromixers 15 are located in adjacent Between the channel inlets of the second-stage Tesla micromixer 17 , the channel outlet of the first-stage Tesla micromixer 15 communicates with the side wall of the second main channel 16 .

As shown in FIG. 2 to FIG. 4 , in order to facilitate the entry of drugs into the first inlet channel 13, a first channel 12 is provided on one side of the upper chip 11. In this embodiment, the cross section of the first channel 12 is L-shaped, and the first channel 12 One end of the first channel 12 penetrates the sidewall of the upper chip 11 , the other end of the first channel 12 penetrates the lower surface of the upper chip 11 , and the other end of the first channel 12 communicates with the first inlet channel 13 .

As shown in FIG. 5 and FIG. 6 , the upper surface of the lower chip 21 is provided with a downwardly recessed cell culture chamber and a first outlet channel 24 , and the number of the cell culture chambers is based on the number of the second-stage Tesla micromixer 17 Setting, the first outlet channel 24 is communicated with the cell culture chamber, the channel outlet of the second-stage Tesla micromixer 17 coincides with the cell culture chamber, and the depth of the cell culture chamber and the depth of the first outlet channel 24 are smaller than those of the lower chip 21. thickness.

In order to study the hepatotoxicity and its liver-related nephrotoxicity under different concentration gradients of drugs, or the liver toxicity and its liver-related nephrotoxicity under different concentrations of two or two drugs, the cell culture room in this example includes a first cell culture room. 22 and the second cell culture room 23, the first cell culture room 22 is communicated with the second cell culture room 23, the number of the first cell culture room 22 is four, the number of the second cell culture room 23 is four, The channel outlets of the second-stage Tesla micromixer 17 coincide with the first cell culture chamber 22 respectively, so that the drug enters the first cell culture chamber 22 from the channel outlet of the second-stage Tesla micromixer 17, and the first outlet One end of the channel 24 is communicated with the second cell culture chamber 23 , and the other end of the first outlet channel 24 is communicated with the outer wall of the lower chip 21 .

As shown in FIG. 3 and FIG. 4 , in order to facilitate the flow of the drug at the channel outlet of the second-stage Tesla micromixer 17 into the first cell culture chamber 22, the channel outlet of the second-stage Tesla micromixer 17 is provided with the first cell culture chamber 22. Two channels 18, the second channel 18 is recessed upward, and the depth of the second channel 18 is smaller than the thickness of the first chip.

The working principle of this embodiment: During the operation, the prepared liver and kidney organ tissues are put into the first cell culture chamber 22 and the second cell culture chamber 23 respectively, and then the upper chip 11 is fixed to the lower chip by screws 21, during operation, drugs of different concentrations enter the first inlet channel 13 from the inlets of the two first channels 12 respectively, and pass through the first main channel 14. Since the first inlet channels 13 are located in two adjacent first-stage tes Pull the middle of the micro-mixer 15 and set it in parallel, the same concentration of the drug enters into two adjacent first-stage Tesla micro-mixers 15, and the first-stage Tesla micro-mixer 15 in the middle has two different concentrations The drug flows into and mixes, the drug concentration in the first-stage Tesla micro-mixer 15 on both sides remains unchanged, and the drug in the first-stage Tesla micro-mixer 15 enters the second main channel 16, from the second main channel 16. The channel 16 flows into the second-stage Tesla micro-mixer 17, and after mixing by the second-stage Tesla micro-mixer 17, four drug concentration gradients are formed. Coincidentally, drugs of different concentrations enter the first cell culture chamber 22 from the channel outlet of the second-stage Tesla micromixer 17 respectively, and then flow through the second cell culture chamber 23 communicated with the first cell culture chamber 22, Finally it flows into the first outlet channel 24 which communicates with the cell culture chamber.

For example, a drug with a concentration of 0 and 2 mol/L is respectively introduced from the inlet of the first channel 12, and after mixing by the three first-stage Tesla micro-mixers, the drug concentration at the channel outlet of the first-stage Tesla micro-mixer They are 0, 1mol/L, and 2mol/L, respectively. After mixing by the second-stage Tesla micromixer, the drug concentrations at the channel outlet of the second-stage Tesla micromixer are 0, 0.5mol/L, and 1.5mol, respectively. /L, 2mol/L, forming four different concentration gradients of drugs.

In this embodiment, the first channel 12, the first inlet channel 13, the first main channel 14, the first-stage Tesla micromixer 15, the second main channel 16, the second-stage Tesla micromixer 17, The widths of the second channel 18 and the first outlet channel 24 are in the micro-scale range (within millimeters), so that the fluid forms a laminar flow.

Beneficial effects of this embodiment: The chip in this embodiment can form a variety of drug concentration gradients, and contains both a hepatocyte chamber and a kidney cell chamber, so that it can directly study the hepatotoxicity and its liver-related effects under different drug concentrations. Nephrotoxicity to improve the efficiency and reliability of drug testing.

The organ drug test chip provided by the invention adopts the Tesla mixing channel, which can be mixed quickly and efficiently to generate a stable drug concentration gradient.

Example 2

The organ drug test chip in Example 1 was used to detect the damage of the drug (acetaminophen) to the liver tissue, the liver tissue was placed in the first cell culture chamber, and the inlets of the first channel 12 were fed with concentrations of 0 and 0.6g respectively. After 24 hours of administration of acetaminophen/L, the cell viability in the first cell culture chamber 22 was detected as shown in Figure 7. The results showed that the drug formed a concentration gradient, and the cell viability increased with the concentration of the drug. increase and decrease, the 50% lethal dose of acetaminophen on hepatocytes shown here is 0.15 g/L.

The above embodiments are only used to illustrate the technical solutions of the present invention, but not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: The recorded technical solutions are modified, or some technical features thereof are equivalently replaced; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims (10)

1. an organ drug testing chip, it is characterized in that: comprise the upper layer chip and the lower layer chip that are arranged sequentially from top to bottom; The bottom surface of the upper chip is provided with at least three recessed Tesla micro-mixers, a first inlet channel and a first main channel, the Tesla micro-mixers are arranged in parallel, and the Tesla micro-mixers are arranged in parallel. The channel inlets are all connected with the first main channel, the first inlet channels are all located in the middle of the adjacent Tesla micromixer, and one end of the first inlet channel is communicated with the first main channel; The upper surface of the lower chip is provided with a recessed cell culture chamber and a first outlet channel, the first outlet channel communicates with the cell culture chamber, and the channel outlet of the Tesla micromixer coincides with the cell culture chamber. 2. The organ drug testing chip according to claim 1, wherein the Tesla micromixer comprises a multistage Tesla micromixer, and the multistage Tesla micromixers are connected in series in sequence connected, the outlet channel of any Tesla micromixer in the previous stage is located in the middle of the inlet channel of the adjacent Tesla micromixer in the subsequent stage, and the number of Tesla micromixers in the latter stage is higher than that in the previous stage. The number of first-stage Tesla micromixers is one more. 3. The organ drug testing chip according to claim 2, wherein the multi-stage Tesla micro-mixer comprises a first-stage Tesla micro-mixer and a second-stage Tesla micro-mixer; The channel inlets of the first-stage Tesla micro-mixers are all connected with the first main channel, and the first inlet channels are all located in the middle of the adjacent first-stage Tesla micro-mixers; The channel inlet of the second-stage Tesla micro-mixer is provided with a second main channel, the channel inlets of the second-stage Tesla micro-mixer are all communicated with the second main channel, and the first-stage Tesla micro-mixer is connected with the second main channel. The channel outlet of the pull micro-mixer is located in the middle of the adjacent second-stage Tesla micro-mixer, and the channel outlet of the second-stage Tesla micro-mixer coincides with the cell culture chamber. 4 . The organ drug testing chip according to claim 3 , wherein a first channel is provided on one side of the upper-layer chip, the cross-section of the first channel is L-shaped, and one end of the first channel passes through the first channel. 5 . Passing through the sidewall of the upper-layer chip, the other end of the first channel penetrates the lower surface of the upper-layer chip, and the other end of the first channel communicates with the first inlet channel. 5. The organ drug testing chip according to claim 2, wherein the number of the first-stage Tesla micro-mixer is three, and the number of the second-stage Tesla micro-mixer is three. is four, and the number of the cell culture chambers is four. 6 . The organ drug test chip according to claim 1 , wherein the cell culture chamber comprises a first cell culture chamber and a second cell culture chamber, and the first cell culture chamber communicates with the second cell culture chamber 6 . , the channel outlet of the Tesla micromixer coincides with the first cell culture chamber, and the second cell culture chamber communicates with one end of the first outlet channel. 7. The organ drug test chip according to claim 6, wherein the number of the first cell culture chamber is the same as the number of the second-stage Tesla micromixer, and the second cell culture chamber The number is the same as the number of the first cell culture chamber. 8 . The organ drug testing chip according to claim 6 , wherein the other end of the first outlet channel is communicated with the outer wall of the lower chip. 9 . 9 . The organ drug testing chip according to claim 1 , wherein the depth of the channel of the Tesla micromixer is smaller than the thickness of the upper chip. 10 . 10 . The organ drug testing chip according to claim 1 , wherein the depth of the cell culture chamber and the depth of the first outlet channel are smaller than the thickness of the lower chip. 11 .

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