Injection and in vivo testing device for VDAs efficacy studies in chicken embryo models

Abstract

The present invention relates to an injection and live detection device for chicken embryo model for VDAs efficacy research, which comprises a container for receiving chicken embryo, an injection mechanism for injecting chorioallantoic membrane blood vessels, and a live detection mechanism for monitoring changes in blood perfusion, wherein the container is provided with a downwardly concave receiving groove, a light-transmitting hole penetrating the container is provided below the container, the light-transmitting hole is communicated with the receiving groove, the injection mechanism comprises a handleless needle, an extension tube, and a micro-injection pump, one end of the extension tube is used as a first connection end, and the other end is used as a second connection end, the second connection end is connected to the syringe in the micro-injection pump, and the first connection end is connected to the handleless needle and forms a clearance fit. The present invention has the following advantages and effects: it can realize micro, accurate, and slow administration of drugs to the chorioallantoic membrane of chicken embryos, and can detect the efficacy of VDAs on vascular changes and blood perfusion in real time and live.

Description

Technical Field

[0001] The invention relates to the technical field of medical devices, and in particular to an injection and in vivo detection device for VDAs efficacy research in a chicken embryo model. Background Art

[0002] Vascular blocking agents, or "VDAs" for short, can constrict blood vessels and reduce blood flow. They can act on normal blood vessels and tumor blood vessels to varying degrees. They take advantage of the difference in their effects on normal blood vessels and pre-formed tumor blood vessels, that is, they preferentially act on the tumor vascular endothelium and destroy its original flat structure, causing the blood vessels to narrow and blood supply to be obstructed, resulting in large-area ischemia and necrosis of the tumor. Thus, they have the advantages of strong tumor inhibition effect, broad anti-tumor spectrum and short action time, and are of great value in tumor treatment.

[0003] At present, the experimental animals used in various preclinical studies of various VDAs are mostly mammals such as mice, rats, rabbits, pigs, and monkeys. However, these animals have certain defects and are increasingly unable to meet the requirements of animal welfare in the future: these experimental animals are convenient for intravascular administration, but the model animals are expensive; even if they can be modeled and completed by themselves, it takes a relatively long time to wait; and the problem of tumor xenotransplantation failure is often encountered in these animal models; although there are some immunodeficient animals available for transplantation, the cost is high, the breeding environment requirements are higher, and the modeling time is very long. In addition, there are generally two ways to observe the effects of VDAs before and after the administration of the above-mentioned experimental animals. One is to use in vitro detection, which will cause irreversible damage to the experimental animals and consume a large number of experimental animals; the other is to use in vivo dynamic detection, such as dynamic enhanced magnetic resonance imaging technology, 19 F magnetic resonance imaging oximetry and dynamic bioluminescence imaging, but these methods are very expensive, difficult to operate, and require high technical skills from the test personnel. Therefore, considering factors such as modeling time, species differences, (animal and testing) costs, and animal welfare, the experimental animals and methods for studying VDAs need to be improved.

[0004] Using chicken embryos instead of mammals for preclinical research on new drugs not only meets animal welfare requirements, but also can detect in vivo changes. It is also a common model organism with low cost, high degree of standardization, short experimental cycle, and easy operation. The chicken embryo chorioallantoic membrane is rich in blood vessels and is a naturally immunodeficient host. The success rate of cross-species tumor cell transplantation is high, and it can be used as an ideal experimental model for vascular pharmacology, oncology, and other studies.

[0005] The traditional chicken embryo chorioallantoic membrane model is mostly used for the study of anti-tumor angiogenesis drugs, and there is almost no study on the efficacy of VDAs on normal blood vessels and tumor blood vessels on the chorioallantoic membrane. There is also a report on the use of chicken embryos to study the efficacy of Compretin disodium phosphate (CA4P, one of the VDAs), but the focus of the study is not on the blocking effect of "formed tumor blood vessels", but on the inhibition of "tumor angiogenesis". Therefore, the use of chorioallantoic membrane blood vessels and their tumor-bearing models to study the efficacy and side effects of VDAs is currently a blank and has room for development.

[0006] However, in the process of implementing the embodiments of the present application, the inventors found that the use of the chicken embryo chorioallantoic membrane model for VDAs efficacy research in the prior art has at least the following problems:

[0007] First, the method of administration. Direct injection of cardiovascular drugs into blood vessels is the most convenient and accurate method of administration. However, due to the relatively small blood vessels of the chicken embryo chorioallantoic membrane model, gelatin sponges, qualitative filter paper or methylcellulose are currently used as carriers in experimental operations. After the drug solution is adsorbed, it is applied to the outside of the chorioallantoic membrane model blood vessels for exchange and absorption. Therefore, the dosage is not accurate enough. If the needle is injected, manual injection is difficult, and the needle may be pulled out due to hand shaking or be deflected by the weight of the needle or the back of the syringe, puncturing the blood vessels. In addition, due to the very limited injectable capacity of the chicken embryo, a small amount of drug cannot be manually administered slowly for a long time. However, if the experimental injection time is shorter than the administration time in actual application, it will bring more obvious side effects to normal blood vessels, thereby failing to correctly simulate the side effects after clinical administration.

[0008] Second, the observation method. When the chorioallantoic membrane model is used for anti-tumor angiogenesis drugs, it is generally photographed in situ in living chicken embryos using a dissecting microscope, but the image can only be used to analyze parameters such as vascular density. The observation of VDAs on the efficacy of tumor blood vessels and side effects on normal blood vessels focuses more on changes in blood perfusion information such as vessel diameter, flow rate, and microvessels. Therefore, the in situ photography method using a dissecting microscope cannot meet the needs of VDAs efficacy research. Summary of the invention

[0009] In view of the shortcomings of the prior art, the object of the present invention is to provide an injection and in vivo detection device for VDAs efficacy research in a chicken embryo model, which can achieve trace, precise and slow drug administration to the chorioallantoic membrane blood vessels, and can detect the efficacy of VDAs on vascular changes and blood perfusion in real time and in vivo.

[0010] To achieve the above object, the present invention provides the following technical solutions:

[0011] The invention discloses an injection and live detection device for chicken embryo model and VDAs efficacy research, characterized in that it comprises a container for receiving chicken embryo, an injection mechanism for chorioallantoic membrane vascular injection and a live detection mechanism for monitoring changes in chorioallantoic membrane blood flow perfusion, the container is provided with a downwardly recessed container groove, a light-transmitting hole penetrating the container is provided below the container, the light-transmitting hole is communicated with the container groove, the injection mechanism comprises a handleless needle, an extension tube and a micro-injection pump, one end of the extension tube serves as a first connection end, and the other end serves as a second connection end, the second connection end is connected to a syringe in the micro-injection pump, the first connection end is connected to the handleless needle and forms a clearance fit, the second connection end is a cast-molded interface, and the interface is sleeved in the syringe in the micro-injection pump.

[0012] The present invention is further configured as follows: the living body detection mechanism includes a laser speckle blood flow imager, an image processing workstation and a bracket, the laser speckle blood flow imager is fixed above the accommodating member through the bracket and is vertically opposite to the accommodating groove, the laser speckle blood flow imager is used to collect chorioallantoic membrane blood flow perfusion images and transmit image information to the image processing workstation, the image processing workstation is used to perform real-time analysis of the blood flow image, and an opening is provided on the side wall of the accommodating member, and the opening is connected to the accommodating groove.

[0013] The present invention is further configured as follows: a heating mechanism is provided at the bottom of the containing tank, the heating mechanism is used to ensure that the chicken embryo is in a normal physiological state, and the heating mechanism is in contact with the bottom of the containing tank.

[0014] The present invention is further configured as follows: the shape of the receiving groove is adapted to the shape of the small end of the chicken embryo and constitutes a limiting fit between the two, and the receiving piece is made of soft silicone material.

[0015] The present invention is further configured as follows: the outer diameter of the handleless needle is 0.3 mm, the extension tube is a thin-walled component with a thickness of 0.5 mm, and the extension tube is made of a flexible material.

[0016] The present invention is further configured such that the bottom of the receiving groove fits with the end of the light-transmitting hole facing the receiving groove.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] Since a container, an injection mechanism and a live detection mechanism are provided, the chicken embryo can be received by the container groove provided in the container, the handleless needle and the flexible extension tube in the injection mechanism prevent the needle from slipping during injection of the chorioallantoic membrane blood vessels and affecting the injection, and the micro-injection pump in the injection mechanism replaces manual injection, thereby improving the injection accuracy and reducing the occurrence of misoperation, and the micro-injection pump can be used to control the injection speed and injection volume before injection, thereby achieving micro-injection and slow injection of the chicken embryo, thereby correctly simulating the side effects after clinical administration, and the live detection mechanism can monitor the blood perfusion in real time during injection. Therefore, the technical problems of low injection accuracy when injecting chicken embryos, failure to inject slowly resulting in failure to correctly simulate the side effects after clinical administration, and failure to meet the observation requirements for VDAs efficacy research are effectively solved. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a schematic diagram of the specific structure of the injection mechanism in the present invention;

[0020] Figure 2 It is a schematic diagram of the positional relationship between the laser speckle blood flow imager and the receiving component in the present invention;

[0021] Figure 3 Schematic diagrams of the axial structure of the accommodation member in the present invention from different viewing angles;

[0022] Figure 4 It is a schematic diagram of the cross-sectional structure of the container in the present invention. DETAILED DESCRIPTION

[0023] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. 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.

[0024] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in 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. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0025] like Figures 1 to 4As shown, the present invention discloses an injection and in vivo detection device for VDAs efficacy research in a chicken embryo model, comprising a container 1 for receiving a chicken embryo, an injection mechanism 2 for chorioallantoic membrane blood vessel injection, and a in vivo detection mechanism for monitoring changes in blood perfusion, wherein the container 1 is provided with a downwardly recessed receiving groove 3, a light transmission hole 4 penetrating the container 1 is provided below the container 1, and the light transmission hole 4 is connected to the receiving groove 3, the injection mechanism 2 comprises a handleless needle 21, an extension tube 22, and a microinjection pump 23, one end of the extension tube 22 is used as a first connection end 221, and the other end is used as a second connection end 22 2, the second connecting end 222 is connected to the syringe 231 in the micro-injection pump 23, the first connecting end 221 is connected to the handleless needle 21 and forms a clearance fit, so as to facilitate the connection between the extension tube 22 and the syringe 231, the second connecting end 222 is a cast-molded interface, and the interface is sleeved in the syringe 231 in the micro-injection pump 23, when the chicken embryo chorioallantoic membrane blood vessels are injected, the chicken embryo can stand upright in the receiving groove 3, and when the chicken embryo is placed in the receiving groove 3, the strong light source can enter the receiving groove 3 through the light transmission hole 4 communicated with the receiving groove 3, so that the blood vessels in the chicken embryo can be clearly displayed, and the blood vessels in the chicken embryo can be clearly displayed. The injection accuracy can be improved, and errors during injection can be effectively prevented. Before injection, the microinjection pump 23 can be used to control the injection speed and injection volume, so that slow constant-current injection of the chicken embryo can be achieved, which solves the problem that the chicken embryo itself can withstand a small injection dose, and avoids the false positive results of VDAs on blood vessels caused by rapid administration, thereby correctly simulating the side effects after clinical administration. After injection, the analysis mechanism can analyze the blood flow changes in real time, thereby facilitating the study of the efficacy of VDAs. When it is necessary to move the chicken embryo to use the living body detection mechanism for more comprehensive observation and analysis, there is no need for manual operation. The handle needle 21 reduces the weight of the needle holding part by removing the handle, thereby preventing the handleless needle 21 from slipping out of the blood vessel during the movement of the chicken embryo, which is convenient for observation and analysis, and the handleless needle 21 and the microinjection pump 23 are connected through the extension tube 22, thereby further improving the movement range of the chicken embryo without the handle needle 21 leaving the blood vessel, which is convenient for observation and analysis, wherein the first connecting end 221 of the extension tube 22 is connected to the handleless needle 21 and forms a clearance fit, thereby facilitating the installation or disassembly of the handleless needle 21 and the extension tube 22, thereby realizing the switching of the drug injection method.

[0026] The living body detection mechanism comprises a laser speckle blood flow imager 5, an image processing workstation and a bracket 6. The laser speckle blood flow imager 5 is fixed on the upper part of the container 1 through the bracket 6 and is vertically opposite to the containing groove 3. The laser speckle blood flow imager 5 is used to collect blood flow images and transmit image information to the image processing workstation. The image processing workstation is used to analyze the blood flow images in real time. An opening 7 is provided on the side wall of the container 1. The opening 7 is connected with the containing groove 3. The laser speckle blood flow imager 5 can analyze the blood flow rate value and the blood vessel diameter more clearly and quickly, and the operation is simple and low in cost. By providing the opening 7 connected with the containing groove 3 on the side wall of the container 1, the side wall of the chicken embryo can be injected, and the laser speckle blood flow imager 5 is placed above the containing groove 3, so that the side wall of the chicken embryo can be injected. At the same time, the laser speckle blood flow imager 5 above the chicken embryo collects blood flow information of the chicken embryo, and then the injection and analysis are performed synchronously, the operation steps are reduced, and the operation is convenient.

[0027] A heating mechanism (not shown) is provided at the bottom of the holding tank 3, and the heating mechanism (not shown) is used to ensure that the chicken embryo is in a normal physiological state. The heating mechanism (not shown) is fitted with the bottom of the holding tank 3, and the chicken embryo is heated by the heating mechanism (not shown) to be in a constant temperature of 37 degrees Celsius, thereby ensuring that the chicken embryo is in a normal physiological state, thereby preventing the chicken embryo from reducing, slowing down, or even stopping metabolism due to hypothermia, causing a change in blood flow, and making the information taken by the analysis mechanism inaccurate.

[0028] The shape of the receiving groove 3 is adapted to the shape of the small end of the chicken embryo and constitutes a limiting fit between the two. The receiving part 1 is made of soft silicone material, so that the receiving groove 3 can fit the chicken embryo more closely, further preventing the chicken embryo from malfunctioning during the injection process and reducing the injection accuracy.

[0029] The outer diameter of the handleless needle 21 is 0.3 mm, and the extension tube 22 is a thin-walled component with a thickness of 0.5 mm. The extension tube 22 is made of a high-temperature resistant flexible material. The handleless needle 21 of this specification is thinner and convenient for piercing the blood vessel and is easy to use. This extension tube 22 is soft in texture and can reduce stress. Therefore, when the extension tube 22 is not straightened, the chicken embryo can be moved arbitrarily and the handleless needle 21 can still remain in the blood vessel, preventing the handleless needle 21 from falling off.

[0030] The bottom of the receiving groove 3 fits with the end of the light-transmitting hole 4 facing the receiving groove 3, thereby ensuring that the strong light source is completely injected into the receiving groove 3, making the blood vessels on the chicken embryo appear more clearly, thereby improving the injection accuracy.

[0031] This specific embodiment is merely an explanation of the present invention and is not a limitation of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. An injection and in vivo detection device for VDAs efficacy research in a chicken embryo model, characterized in that: The invention comprises a container for receiving a chicken embryo, an injection mechanism for injecting blood vessels of the chorioallantoic membrane of the chicken embryo, and a living body detection mechanism for monitoring changes in blood perfusion of the chorioallantoic membrane. The container is provided with a downwardly concave receiving groove, and an opening communicating with the receiving groove is provided on the side wall of the container, which is used for injecting the side wall of the chicken embryo. A light-transmitting hole penetrating the accommodating member is provided below the accommodating member, and the light-transmitting hole is communicated with the accommodating groove. The injection mechanism includes a handleless needle, an extension tube and a micro-injection pump. The extension tube is made of a high-temperature resistant flexible material. One end of the extension tube serves as a first connecting end, and the other end serves as a second connecting end. The second connecting end is connected to the syringe in the micro-injection pump. The first connecting end is connected to the handleless needle and forms a clearance fit. The second connecting end is a cast interface, and the interface is mounted in the syringe in the micro-injection pump.

2. The injection and in vivo detection device for VDAs efficacy research in a chicken embryo model according to claim 1, characterized in that: The living body detection mechanism includes a laser speckle blood flow imager, an image processing workstation and a bracket. The laser speckle blood flow imager is fixed above the receiving member through the bracket and is vertically opposite to the receiving slot. The laser speckle blood flow imager is used to collect chorioallantoic membrane blood flow perfusion images and transmit image information to the image processing workstation. The image processing workstation is used to perform real-time analysis of blood flow images. An opening is provided on the side wall of the receiving member, and the opening is connected to the receiving slot.

3. The injection and in vivo detection device for VDAs efficacy research in a chicken embryo model according to claim 1, characterized in that: A heating mechanism is provided at the bottom of the containing tank, and the heating mechanism is used to ensure that the chicken embryo is in a normal physiological state. The heating mechanism is in contact with the bottom of the containing tank.

4. The injection and in vivo detection device for VDAs efficacy research in a chicken embryo model according to claim 3, characterized in that: The bottom of the receiving groove is in contact with the end of the light-transmitting hole facing the receiving groove.

5. The injection and in vivo detection device for VDAs efficacy research in a chicken embryo model according to claim 1, characterized in that: The shape of the receiving groove is adapted to the shape of the small end of the chicken embryo and constitutes a limiting fit between the two, and the receiving piece is made of soft silicone material.

6. The injection and in vivo detection device for VDAs efficacy research in a chicken embryo model according to claim 1, characterized in that: The outer diameter of the handleless needle is 0.3 mm, the extension tube is a thin-walled component with a thickness of 0.5 mm, and the extension tube is made of a flexible material.

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