A three-dimensional microscopic bone lacuna-tubule system mass transfer chip and experimental device

Abstract

The present invention patent discloses a three-dimensional microscopic bone lacuna-tubule system mass transfer chip and experimental device, specifically relating to the fields of organoids, microfluidics, and bone tissue engineering devices. The mass transfer chip includes an upper cover plate, multiple intermediate plates, and a lower cover plate. The upper cover plate is connected to adjacent intermediate plates, adjacent intermediate plates, and the intermediate plate to the lower cover plate by bonding. The centers of the upper cover plate, multiple intermediate plates, and lower cover plate are all pierced with Haversian tubes. Each intermediate plate is etched with multiple bone lacunas spaced in an annular pattern. Bone canaliculi connect two adjacent bone lacunas. The bone canaliculi connect the bone lacunas to the Haversian tubes. The Haversian tubes, bone lacunas, and bone canaliculi are all made of quartz glass. The technical solution of the present invention solves the problems of existing bone mass transfer experimental research and can examine the transport process of the bone lacuna-tubule system.

Description

Technical Field

[0001] The present invention relates to the field of ray shielding devices, and in particular to a three-dimensional microscopic bone lacuna-tubule system mass transfer chip and an experimental device. Background Art

[0002] Nutrient delivery and waste elimination are the basic conditions for the metabolism of organisms. The mass transfer process from capillaries to bone cells requires interstitial transport through the lacunae-tubule system to achieve material exchange. The lacunae-tubule system is the mechanobiological basis of bone tissue metabolism, mechanical conduction and adaptation.

[0003] Currently, scholars often use in vivo or in vitro experiments to explore the mass transfer laws of the bone lacuna-tubule system, which requires the use of bone in vivo as the experimental subject. However, the bones in different organisms are different, and it is impossible to conduct simulation experiments by precisely controlling a single variable, which will affect the experimental data. At present, the research on organoids, microfluidics and bone tissue engineering is developing rapidly. There is an urgent need to explore the material transfer process inside human tissue, especially the mass transfer laws in the microscopic pores of bone. In addition, the mass transfer process of bone extracted from the body is very complex, and the technology is not yet available. It cannot be experimentally studied in extreme environments (such as in the microgravity of an orbiting space station).

[0004] With the advancement of microfabrication technology, laser etching can now create organoid chips with customized microstructures. Organoid chips can simulate real physiological environments, providing a more efficient experimental platform. Compared to traditional biological experiments, organoid chips offer more precise and controllable experimental conditions, reducing experimental errors. Furthermore, using organoid chips for experiments presents no ethical issues.

[0005] Therefore, how to use organoid chips for mass transfer experiments in the bone crypt-tubule system has become a difficult problem that needs to be solved urgently. Summary of the Invention

[0006] The present invention aims to provide a three-dimensional microscopic bone lacuna-tubule system mass transfer chip and experimental device, which solves the problems of existing bone mass transfer experimental research.

[0007] To achieve the above objectives, the present invention provides a technical solution as follows: a three-dimensional microscopic bone lacuna-tubule system mass transfer chip, comprising an upper cover plate, multiple intermediate plates, and a lower cover plate, wherein the upper cover plate is connected to adjacent intermediate plates, adjacent intermediate plates are connected to each other, and the intermediate plate is connected to the lower cover plate by bonding, and the centers of the upper cover plate, the multiple intermediate plates, and the lower cover plate are all penetrated by a Haversian tube, each of the intermediate plates is etched with a plurality of annularly spaced bone lacunas, and bone canaliculi are connected between two adjacent bone lacunas, and all of the bone canaliculi connect the bone lacunas to the Haversian tube, and the Haversian tube, bone lacuna, and bone canaliculi are all made of quartz glass;

[0008] Bone canaliculi are connected between the bone cavities at the same position on two adjacent intermediate plates;

[0009] The Haversian tube has a diameter of 50 μm and a depth of 300 μm;

[0010] The bone lacuna has a long diameter of 10 μm, a short diameter of 5 μm, and a depth of 5 μm;

[0011] The diameter of the bone canaliculus is 0.7 μm.

[0012] Through the above settings, the entire mass transfer system has a three-dimensional structure, which can more realistically simulate the entire process of bone mass transfer in the body. At the same time, the use of real dimensions makes the simulated data more realistic and accurate.

[0013] Another technical solution provided by the present invention is as follows: an experimental device for a three-dimensional microscopic bone lacuna-tubule system mass transfer chip, comprising any of the mass transfer chips described above, wherein both ends of the Haversian tube in the mass transfer chip are connected to connecting tubes, each of the connecting tubes is connected to a liquid accumulation tank, a tracer is provided in any of the liquid accumulation tanks, and a buffer solution is injected into the other liquid accumulation tank, and the mass transfer chip and the liquid accumulation tank are jointly connected to a centrifuge.

[0014] Furthermore, the experimental setup of the mass transfer chip is as follows:

[0015] S1. Connect both ends of the Haversian tube in the mass transfer chip to the liquid accumulation tank through connecting tubes;

[0016] S2, injecting a tracer into any one of the fluid collection tanks in step S1, and injecting a buffer solution into the other fluid collection tank;

[0017] S3, placing the two liquid collection tanks injected with the tracer and buffer in step S2, the connecting tube and the mass transfer chip together on a centrifuge;

[0018] S4. Use a centrifuge to drive the effusion tank and mass transfer chip to rotate. By adjusting the speed of the centrifuge to simulate different gravity environments, the fluorescent tracer flows from the effusion tank into the bottom of the three-dimensional microscopic bone lacuna-tubule system mass transfer chip and is injected for 20 minutes. Stop loading; remove the three-dimensional microscopic bone lacuna-tubule system mass transfer chip, place it under a laser confocal microscope for observation, take images, record experimental data, and analyze the mass transfer rules in the mass transfer chip.

[0019] Furthermore, the two liquid accumulation tanks, the connecting pipe and the mass transfer chip are collectively covered with a silicone sleeve.

[0020] Through the above arrangement, the silicone sleeve can be used to form the liquid accumulation tank, the connecting tube and the mass transfer chip into a whole, which is beneficial to the stability of the mass transfer process during the experiment.

[0021] Compared with the existing technology, this solution has the following beneficial effects:

[0022] This three-dimensional microscopic lacunae-canaliculi mass transfer chip, built on the spatial structure of bone units, accurately simulates the transport process within the lacunae-canaliculi system. This microscopic lacunae-canaliculi mass transfer chip provides a fundamental foundation for mass transfer experimental research. Furthermore, this solution can also be used for experimental research in extreme environments, such as the microgravity of an orbiting space station. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is an axonometric view of a three-dimensional microscopic bone lacuna-tubule system mass transfer chip in Example 1;

[0024] Figure 2 is a schematic structural diagram of the upper cover plate in Example 1;

[0025] Figure 3 is a schematic structural diagram of the middle plate in Example 1;

[0026] Figure 4 Schematic diagram of the mass transfer system structure on the middle plate in Example 1;

[0027] Figure 5 It is a partial enlarged schematic diagram of the mass transfer system in Example 1;

[0028] Figure 6 is a schematic diagram of the mass transfer chip of Example 1 during an experiment;

[0029] Figure 7 This is a visualization diagram of the mass transfer of middle molecular particles in a bone lacuna under microgravity in the mass transfer chip of Example 1;

[0030] Figure 8 This is a visualization diagram of the mass transfer of middle molecular particles in a bone lacuna under normal gravity in the mass transfer chip of Example 1;

[0031] Figure 9 This is a visualization diagram of the mass transfer of intermediate molecular particles in a bone lacuna under high gravity (10g) in the mass transfer chip of Example 1. DETAILED DESCRIPTION

[0032] The present invention will be further described in detail below through specific embodiments:

[0033] The reference numerals in the drawings of the specification include: upper cover plate 1, middle plate 2, lower cover plate 3, Haversian tube 4, bone lacuna 5, bone canaliculus 6, connecting tube 7, fluid accumulation tank 8, mass transfer chip 9, silicone sleeve 10, centrifuge 11.

[0034] Example 1

[0035] like Figures 1 to 6As shown, a three-dimensional microscopic bone lacuna-tubule system mass transfer chip comprises a circular upper cover plate 1, multiple intermediate plates 2, and a lower cover plate 3. In this embodiment, there are ten intermediate plates 2, and the upper cover plate 1, all intermediate plates 2, and lower cover plates 3 are made of quartz glass. The upper cover plate 1 is connected to adjacent intermediate plates 2, to adjacent intermediate plates 2, and to the lower cover plate 3 via bonding. A Haversian tube 4 is provided through the center of the upper cover plate 1, all intermediate plates 2, and lower cover plate 3. The Haversian tube 4 has a diameter of 50 μm and a depth of 300 μm. Each intermediate plate 2 is etched with multiple bone lacunas 5 of varying diameters, spaced apart in an annular pattern. The lacunas 5 have a major diameter of 10 μm, a minor diameter of 5 μm, and a depth of 5 μm. Between two adjacent lacunae 5, a canaliculus 6 is connected. The canaliculus 6 has a diameter of 0.7 μm. All canaliculi 6 connect the lacunae 5 to the Haversian tubules 4. The Haversian tubules 4, lacunae 5, and canaliculi 6 are all made of quartz glass. In this embodiment, the structural dimensions of the Haversian tubules, lacunae 5, and canaliculi 6 are identical to those found in real bone structures, effectively simulating the mass transfer process of bone in vivo.

[0036] In this embodiment, each intermediate plate 2 has eight radially distributed canaliculi 6 connected to the Haversian canal at the center. Each radially arranged canaliculus 6 has four bone lacunae 5. The bone lacunae 5 are connected to each other through the canaliculi 6. Each bone lacunae 5 is etched with a canaliculus 6 that is connected to the bone lacunae 5 at the same position on the next intermediate plate 2, thus forming a three-dimensional bone lacunae 5-canaliculus mass transfer system.

[0037] Finite element simulation shows that the mass transfer chip of the three-dimensional microscopic bone lacuna-tubule system can realize the mass transfer process (such as Figure 7-9 Figures 2 and 3 show simulations of a mass transfer chip under different gravity fields, and numerical analyses of mass transfer patterns under microgravity, normal gravity, and hypergravity. The spheres represent pits, the spaces between the spheres are connected by canaliculi, and the particles within the pits represent mass transfer. The simulations demonstrate that hypergravity promotes mass transfer within the model. Experimental studies of mass transfer under microgravity, normal gravity, and hypergravity are feasible.

[0038] Example 2

[0039] An experimental device for a three-dimensional microscopic bone lacuna 5-tubule system mass transfer chip 9 includes the mass transfer chip 9 of Example 1. Both ends of the Haversian tube 4 in the mass transfer chip 9 are connected to connecting tubes 7, and each connecting tube 7 is connected to a liquid accumulation tank 8. A tracer is provided in any liquid accumulation tank 8, and a buffer solution is injected into the other liquid accumulation tank 8. The mass transfer chip 9 and the liquid accumulation tank 8 are jointly covered with a silicone sleeve 10. A placement groove for placing the mass transfer chip 9 and the liquid accumulation tank 8 is provided in the silicone sleeve 10. The mass transfer chip 9 and the liquid accumulation tank 8 are connected to a centrifuge 11 through the silicone sleeve 10.

[0040] The experimental method of the experimental device of this embodiment is as follows:

[0041] S1. Connect both ends of the Haversian tube 4 in the mass transfer chip 9 to the liquid storage tank 8 through the connecting tube 7;

[0042] S2, injecting a tracer into any one of the liquid collection tanks 8 in step S1. In this embodiment, a small molecule tracer is used as the tracer, and injecting a buffer solution into the other liquid collection tank 8;

[0043] S3. Inject the tracer and buffer from step S2 into the two liquid collection tanks 8, respectively. Place the liquid collection tanks 8, the connecting tube 7, and the mass transfer chip 9 together on a centrifuge 11 through a silicone sleeve 10. In this embodiment, multiple experimental devices can be placed on the centrifuge 11.

[0044] S4. Use the centrifuge 11 to drive the two liquid accumulation tanks 8 and the mass transfer chip 9 to rotate. By adjusting the speed of the centrifuge 11 to simulate different gravity environments, the fluorescent tracer flows from the liquid accumulation tank 8 into the bottom of the three-dimensional microscopic bone lacuna 5-tubule system mass transfer chip 9 and is injected for 20 minutes. Stop loading; take out the three-dimensional microscopic bone lacuna 5-tubule system mass transfer chip 9, place it under a laser confocal microscope for observation, take images, record experimental data, and analyze the mass transfer rules in the mass transfer chip 9.

[0045] Example 3

[0046] The only difference between this embodiment and embodiment 2 is that in this embodiment, there are three intermediate plates 2 , and the rest of the structure is the same as that of embodiment 2.

[0047] The difference between the experimental device of this embodiment and the experimental device of embodiment 2 is that in this embodiment, a middle molecule tracer is used as the tracer.

[0048] Example 4

[0049] The only difference between this embodiment and embodiment 2 is that in this embodiment, the number of intermediate plates 2 is twenty, and the rest of the structure and steps are the same as those in embodiment 2.

[0050] The experimental apparatus of this embodiment differs from that of embodiment 2 only in that in this embodiment, a macromolecular tracer is used as the tracer, and the remaining steps are the same as those of embodiment 2.

[0051] The above are only embodiments of the present invention, and common knowledge such as the specific structure and / or characteristics of the scheme are not described in detail here. It should be pointed out that for those skilled in the art, without departing from the structure of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A three-dimensional microscopic bone lacuna-tubule system mass transfer chip, characterized by: The invention comprises an upper cover plate (1), a plurality of intermediate plates (2) and a lower cover plate (3), wherein the upper cover plate (1) is connected to adjacent intermediate plates (2), adjacent intermediate plates (2) and the intermediate plates (2) and the lower cover plate (3) by bonding, and the centers of the upper cover plate (1), the plurality of intermediate plates (2) and the lower cover plate (3) are all provided with a Haversian tube (4), and each intermediate plate (2) is etched with a plurality of bone pits (5) distributed in an annular manner, and bone canaliculi (6) are connected between two adjacent bone pits (5), and all the bone canaliculi (6) connect the bone pits (5) with the Haversian tube (4), and the Haversian tube (4), the bone pits (5) and the bone canaliculi (6) are all made of quartz glass; Bone canaliculi (6) are connected between the bone fossa (5) at the same position on two adjacent intermediate plates (2); The Haversian tube (4) has a diameter of 50 μm and a depth of 300 μm; The bone lacuna (5) has a long diameter of 10 μm, a short diameter of 5 μm, and a depth of 5 μm; The diameter of the bone canaliculus (6) is 0.7 μm.

2. A three-dimensional microscopic bone lacuna-tubule system mass transfer chip experimental device, characterized by: The mass transfer chip (9) comprises the mass transfer chip (9) according to claim 1, wherein both ends of the Haversian tube (4) in the mass transfer chip (9) are connected to connecting tubes (7), each connecting tube (7) is connected to a liquid accumulation tank (8), a tracer is provided in any one of the liquid accumulation tanks (8), and a buffer solution is injected into the other one of the liquid accumulation tanks (8), and the mass transfer chip (9) and the liquid accumulation tank (8) are commonly connected to a centrifuge (11).

3. The experimental device for a three-dimensional microscopic bone lacuna-tubule system mass transfer chip according to claim 2, characterized in that: Experimental method of the experimental device: S1. Connect both ends of the Haversian tube (4) in the mass transfer chip (9) to the liquid accumulation tank (8) through the connecting tube (7); S2, injecting a tracer into any one of the liquid collection tanks (8) in step S1, and injecting a buffer solution into the other liquid collection tank (8); S3, placing the two liquid accumulating tanks (8) injected with the tracer and the buffer solution in step S2, the connecting tube (7) and the mass transfer chip (9) together on a centrifuge (11); S4. The centrifuge (11) is used to drive the effusion tank (8) and the mass transfer chip (9) to rotate. By adjusting the rotation speed of the centrifuge (11) to simulate different gravity environments, the fluorescent tracer flows from the effusion tank (8) into the bottom of the three-dimensional microscopic bone cavities (5)-tubule system mass transfer chip (9) and is injected for 20 minutes. The loading is stopped. The three-dimensional microscopic bone cavities (5)-tubule system mass transfer chip (9) is taken out and placed under a laser confocal microscope for observation. Images are taken, experimental data are recorded, and the mass transfer rules in the mass transfer chip (9) are analyzed.

4. The experimental device for a three-dimensional microscopic bone lacuna-tubule system mass transfer chip according to claim 3, characterized in that: The two liquid accumulation tanks (8), the connecting pipe (7) and the mass transfer chip (9) are collectively covered with a silica gel sleeve (10).

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