Three-dimensional microcosmic bone fossa-tubule system mass transfer chip and experimental device

Abstract

The invention discloses a three-dimensional microcosmic bone fossa-tubule system mass transfer chip and an experimental device, and particularly relates to the field of organoids, microfluidics and bone tissue engineering devices. The mass transfer chip comprises an upper cover plate, a plurality of middle plates and a lower cover plate, the upper cover plate is connected with the adjacent middle plates in a bonding mode, the adjacent middle plates are connected in a bonding mode, the middle plates are connected with the lower cover plate in a bonding mode, and the centers of the upper cover plate, the multiple middle plates and the lower cover plate are jointly provided with Harvard pipes in a penetrating mode. A plurality of bone pits which are annularly distributed at intervals are etched in each middle plate, a bone tubule is communicated between every two adjacent bone pits, the bone pits are communicated with the Harvard tube through the bone tubules, and the Harvard tube, the bone pits and the bone tubules are all made of quartz glass. By the adoption of the technical scheme, the problem of existing bone mass transfer experiment research is solved, and the transportation process of a bone fossa-tubule system can be checked.

Description

Technical Field

[0001] The 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 osteocytes 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] At present, scholars often use in vivo or in vitro experiments to explore the mass transfer rules of the bone lacuna-tubule system, which requires the use of bones in vivo as experimental subjects. 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, and there is an urgent need to explore the material transfer process inside human tissues, especially the mass transfer rules in the microscopic pores of bones. In addition, the mass transfer process of extracting bones from organisms is very complicated, and the technology is not up to standard. It cannot be experimentally studied in extreme environments (such as microgravity in orbiting space stations).

[0004] Nowadays, with the development of micro-processing technology, organoid chips with specific microstructures can be processed through laser etching technology. Organoid chips can simulate the real physiological environment and provide a more effective experimental platform. Compared with traditional biological experiments, organoid chips can provide more precise and controllable experimental conditions and reduce experimental errors. In addition, using organoid chips for experiments does not involve ethical issues.

[0005] Therefore, how to use organoid chips for mass transfer experiments in the bone lacunae-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] In order to achieve the above-mentioned purpose, 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, a plurality of intermediate plates and a lower cover plate, wherein the upper cover plate is connected to adjacent intermediate plates, between adjacent intermediate plates, and between the intermediate plate and the lower cover plate by bonding, and the centers of the upper cover plate, the plurality of intermediate plates and the lower cover plate are commonly penetrated with a Haversian tube, each of the intermediate plates is etched with a plurality of bone lacunas distributed in an annular manner, bone canaliculi are connected between two adjacent bone lacunas, and all the bone canaliculi connect the bone lacunas with the Haversian tubes, and the Haversian tubes, bone lacunas and bone canaliculi are all made of quartz glass;

[0008] Bone canaliculi are connected between the bone depressions at the same position on two adjacent middle 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 arrangement, the entire mass transfer system has a three-dimensional structure, which can more realistically simulate the entire process of bone mass transfer in vivo. 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 of the liquid accumulation tanks, and the mass transfer chip and the liquid accumulation tank are commonly connected to a centrifuge.

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

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

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

[0017] S3, placing the two liquid storage tanks injected with the tracer and the 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 the mass transfer chip to rotate, and adjust 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; take out the three-dimensional microscopic bone lacuna-tubule system mass transfer chip, observe it under a laser confocal microscope, 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 coated 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 prior art, this solution has the following beneficial effects:

[0022] The three-dimensional microscopic bone lacuna-tubule mass transfer chip of this scheme has the spatial structure of bone units, which can accurately simulate the transport process of the bone lacuna-tubule system. The microscopic bone lacuna-tubule system mass transfer chip established can solve the basic equipment for mass transfer experimental research. At the same time, this scheme can also be used for experimental research in extreme environments (such as microgravity conditions in 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 is a schematic diagram of the mass transfer system structure on the middle plate in Example 1;

[0027] Figure 5 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 when conducting 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 mass transfer of middle molecular particles in a bone lacuna under normal gravity in the mass transfer chip of Example 1;

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

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

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

[0034] Example 1

[0035] like Figures 1 to 6As shown, a three-dimensional microscopic bone lacuna-tubule system mass transfer chip includes an upper cover plate 1, multiple intermediate plates 2 and a lower cover plate 3, all of which are circular in shape. In this embodiment, the number of intermediate plates 2 is ten, wherein the upper cover plate 1, all intermediate plates 2 and lower cover plate 3 are made of quartz glass. The upper cover plate 1 and adjacent intermediate plates 2, between adjacent intermediate plates 2, and between intermediate plates 2 and lower cover plate 3 are connected together by bonding. The center of the upper cover plate 1, all intermediate plates 2 and lower cover plate 3 is commonly penetrated by a Haversian tube 4, the diameter of the Haversian tube 4 is 50μm and the depth is 300μm. Each intermediate plate 2 is etched with a plurality of bone lacunas 5 with different diameters and annular spacing, and the long diameter of the bone lacuna 5 is 10μm, the short diameter is 5μm, and the depth is 5μm. A canaliculus 6 is connected between two adjacent bone lacunas 5, and the diameter of the canaliculus 6 is 0.7 μm. All canaliculus 6 connect the bone lacunas 5 with the Haversian tubes 4. The Haversian tubes 4, the bone lacunas 5 and the canaliculus 6 are all made of quartz glass. In this embodiment, the structural dimensions of the Haversian tubes, the bone lacunas 5 and the canaliculus 6 are the same as those in the real bone structure, and can truly simulate the mass transfer process of the bone in vivo.

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

[0037] The mass transfer chip of this invention has been demonstrated through finite element simulation 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 The simulations of mass transfer chips under different gravity fields are shown in Figure 1, respectively. The numerical analysis of mass transfer laws under microgravity, normal gravity and hypergravity are shown in Figure 2. The spheres are pits, the spheres are connected by canaliculi, and the particles in the pits are descriptions of mass transfer. The simulations show that the hypergravity environment promotes the mass transfer process in the model. Experimental studies on 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, 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, any liquid accumulation tank 8 is provided with a tracer, and the other liquid accumulation tank 8 is injected with a buffer solution, the mass transfer chip 9 and the liquid accumulation tank 8 are jointly coated with a silicone sleeve 10, the silicone sleeve 10 is provided with a placement groove for placing the mass transfer chip 9 and the liquid accumulation tank 8, and 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, connecting 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 another liquid collection tank 8;

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

[0044] S4. The two liquid accumulation tanks 8 and the mass transfer chip 9 are driven to rotate by the centrifuge 11. The rotation speed of the centrifuge 11 is adjusted to simulate different gravity environments. The fluorescent tracer flows from the liquid accumulation tank 8 into the three-dimensional microscopic bone lacuna 5-tubule system mass transfer chip 9 and is injected at the bottom. The loading is performed for 20 minutes and the loading is stopped. The three-dimensional microscopic bone lacuna 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.

[0045] Example 3

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

[0047] The difference between the experimental device of this embodiment and the experimental device of embodiment 2 is that in this embodiment, the tracer is a middle molecule 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 difference between the experimental device of this embodiment and the experimental device of embodiment 2 is 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 the common knowledge such as the known specific structures and / or characteristics in the scheme are not described in detail here. It should be pointed out that for those skilled in the art, several deformations and improvements can be made without departing from the structure of the present invention, which should also be regarded as the protection scope 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 in that: The invention comprises an upper cover plate (1), a plurality of intermediate plates (2) and a lower cover plate (3); the upper cover plate (1) and adjacent intermediate plates (2), adjacent intermediate plates (2) and intermediate plates (2) and lower cover plate (3) are connected by bonding; a Haversian tube (4) is provided in the center of the upper cover plate (1), the plurality of intermediate plates (2) and lower cover plate (3); a plurality of bone recesses (5) are etched on each intermediate plate (2) and are distributed in an annular manner; bone canaliculi (6) are connected between two adjacent bone recesses (5); all the bone canaliculi (6) connect the bone recesses (5) with the Haversian tube (4); the Haversian tube (4), the bone recesses (5) and the bone canaliculi (6) are all made of quartz glass; Bone canaliculi (6) are connected between the bone lacunas (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. An experimental device for a three-dimensional microscopic bone lacuna-tubule system mass transfer chip, characterized in that: It comprises a mass transfer chip (9) as described in any one of claims 1 to 5, wherein both ends of the Haversian tube (4) in the mass transfer chip (9) are connected to connecting tubes (7), each of the connecting tubes (7) is connected to a liquid collection tank (8), a tracer is provided in any of the liquid collection tanks (8), and a buffer solution is injected into another of the liquid collection tanks (8), and the mass transfer chip (9) and the liquid collection 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, connecting both ends of the Haversian tube (4) in the mass transfer chip (9) to the liquid storage tank (8) through the connecting tube (7); S2, injecting a tracer into any one of the liquid collection tanks (8) of step S1, and injecting a buffer into another liquid collection tank (8); S3, placing the two liquid storage tanks (8) injected with the tracer and the buffer 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 fluid accumulation tank (8) and the mass transfer chip (9) to rotate. The rotation speed of the centrifuge (11) is adjusted to simulate different gravity environments. The fluorescent tracer flows from the fluid 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. The loading is stopped. The three-dimensional microscopic bone lacuna (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. An 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 coated with a silica gel sleeve (10).

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