JP4169376B2 - Tissue cell pressure sensitivity tester - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a pressure sensitivity test apparatus for biological tissues used for pressure sensitivity tests of tissue cells such as vascular endothelial cells and smooth muscle cells, and more particularly to a pressure sensitivity test apparatus for biological tissues that can perform various pressure sensitivity tests. It is.
[0002]
[Prior art]
In recent years, in order to elucidate the mechanism of pathology and study pharmacological effects, it senses the pressure received by surrounding tissue cells of bones, organs, nerves, blood vessels, etc., and shows various responses to this pressure. In order to test the pressure sensitivity of this living tissue, several test devices have been proposed.
[0003]
In the initial stage, as shown in FIG. 3, a device for applying pressure to the tissue cell 101 by directly placing the weight 102 on the tissue cell 101 was used. In this device, the tissue cell 101 is uniformly pressurized. There is doubt about the reliability of the test results.
[0004]
Therefore, as shown in FIG. 4, tissue cells are accommodated in a flask 201, and a pressure gauge (mercury sphygmomanometer) 203, a helium gas cylinder 204, and a pressure setting valve 205 are placed in parallel in the flask 201 via a three-way valve 202. A tissue cell pressure sensitivity test apparatus is used which is connected so that a uniform pressure can be applied without applying shear stress or tension to tissue cells.
[0005]
[Problems to be solved by the invention]
However, these conventional tissue cell pressure sensitivity test devices cannot test the pressure sensitivity of tissue cells to pressures lower than atmospheric pressure, and are sensitive to pressures with periodic fluctuations such as blood pressure. Nor can tissue cell pressure sensitivity be tested.
[0006]
The present invention relates to a tissue cell pressure sensitivity test apparatus capable of performing various pressure sensitivity tests such as a pressure sensitivity test of tissue cells against a pressure lower than atmospheric pressure and a pressure sensitivity test of tissue cells against a periodically changing pressure. It is intended to provide.
[0007]
[Means for Solving the Problems]
The present invention includes a volume-changing culture chamber that contains and seals a sample of body tissue, a pressurizing unit that introduces a gas into the culture chamber at a predetermined introduction pressure, and an opening / closing unit that is connected to the culture chamber. with the atmosphere communication passage connected, and a vacuum pump for reducing the pressure of which is connected to the culture chamber said culture chamber, said pressurizing means, said atmosphere communicating passage, the connection between the vacuum pump and the culture chamber The pressure in the culture chamber is changed, and the pressure applied to the sample is changed.
[0008]
Therefore, according to the first aspect of the present invention, the gas can be introduced at a predetermined introduction pressure by the pressurizing means , so that the internal pressure of the culture chamber can be set to a predetermined introduction pressure of atmospheric pressure or higher. A tissue cell pressure sensitivity test can be performed.
[0011]
Further, according to the first aspect of the present invention, gas can be derived from the culture chamber whose volume does not change, or the internal pressure of the culture chamber can be set to a predetermined pressure of atmospheric pressure or less with a vacuum pump . Tissue cell pressure sensitivity test can be performed.
[0012]
Further, according to the first invention, since both the pressurizing function and the decompression function are provided, the internal pressure of the culture chamber can be freely changed. For example, the pressurization function and the decompression function are alternately and periodically. By acting, it is possible to periodically change the internal pressure of the culture chamber, and to test the sensitivity of the tissue cell pressure to the periodically changing pressure.
[0014]
In addition, the culture chamber has a gas volume variably formed, a pressurizing means for reducing the gas volume of the culture chamber to increase its internal pressure, and expanding the gas volume of the culture chamber to increase its internal pressure. It can comprise so that the pressure reduction means to reduce may be provided .
[0015]
After the sample is placed in the culture chamber at atmospheric pressure and the culture chamber is sealed, the pressure sensitivity of the tissue cells above atmospheric pressure can be tested by reducing the volume of the gas in the culture chamber. It is possible to test the pressure sensitivity of tissue cells below atmospheric pressure by enlarging the volume.
[0016]
In the above, as a specific configuration for making the gas volume variable in the culture chamber, the culture chamber is configured with a cylinder and a piston that is slidable inside the cylinder and is airtightly fitted therein, A part or all of the culture chamber may be formed of an elastic body. When the culture chamber is formed of an elastic body, it is preferable to form the culture chamber in a bellows shape so that it can be deformed greatly without difficulty.
[0017]
In the above, the culture chamber may be directly expanded / reduced by manually pushing and pulling a part of the culture chamber, but in order to accurately control the volume expansion rate and / or the volume reduction rate, the pressurizing means is used. At least one of the pressure reducing means and the pressure reducing means is preferably constituted by a mechanical mechanism that mechanically expands and contracts the culture chamber, and more preferably, the pressure means and the pressure reducing means are constituted by a mechanical mechanism.
[0018]
These pressurizing means and pressure reducing means may be provided independently of each other, but in order to reduce the number of parts, simplify the configuration, and reduce costs, for example, a slider crank mechanism, a crank mechanism, a cam mechanism, It is advantageous to integrate the pressurizing unit and the depressurizing unit using a mechanism that reciprocates like a pantograph mechanism. Among these, in order to be able to adjust the internal pressure of the culture chamber in various ways, the volume change rate can be adjusted by adjusting the position of the connecting contact between the connecting rod connecting the culture chamber to the crank arm and the crank arm. It is recommended to use a slider crank mechanism or crank mechanism that can be adjusted.
[0019]
Furthermore, in the tissue cell pressure sensitivity test apparatus according to the second invention , in addition to the feature of the first invention, a space for changing the volume of gas is communicated with the culture chamber, and the volume of gas in the space is reduced. A pressurizing means for increasing the internal pressure of the culture chamber and a decompression means for reducing the internal pressure of the culture chamber by expanding the volume of gas in the space.
[0020]
According to the second aspect of the present invention , after the sample is accommodated in the culture chamber at atmospheric pressure and the culture chamber is sealed, the internal pressure of the culture chamber is increased from the atmospheric pressure by reducing the volume of the gas in the space. It is possible to test the pressure sensitivity of tissue cells above atmospheric pressure. Further, by expanding the volume of the gas in the space, the internal pressure of the culture chamber can be lowered below atmospheric pressure, and the pressure sensitivity of tissue cells below atmospheric pressure can be tested.
[0021]
In addition, in this way, in addition to the culture chamber, a space that can be expanded and contracted is provided, and when the internal pressure of the culture chamber is controlled by expanding and contracting this space, even if the space is expanded and contracted periodically, Since mechanical vibration accompanying expansion and contraction is difficult to be transmitted to the culture chamber, the culture chamber is hardly vibrated to cause the sample to exhibit pressure sensitivity to the vibration, and a highly reliable test can be performed.
[0022]
In the second invention, in the space variable volume Specifically, slidably inside and this cylinder, and then fitted to the piston airtightly, composed space defined in the cylinder by the piston Or a part or all of the space in the container formed of an elastic body. In the case where this space is partially or entirely formed in a container formed of an elastic body , it is preferable to use a bellows formed as a bellows as an elastic body so that the space can be expanded and contracted without difficulty. .
[0023]
Detailed description of the pressurizing means and the pressure reducing means of the present second invention, since the overlap with that of the previous SL pressurization means and decompression means, is omitted here in order to save paper width.
[0024]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, a tissue cell pressure sensitivity test apparatus according to an embodiment of the present invention will be described with reference to the drawings.
[0025]
As shown in the block diagram of FIG. 1, the tissue cell pressure sensitivity test apparatus includes a culture chamber 1 that does not change volume, and a pressurized air introduction path 2 that introduces air into the culture chamber 1 at a predetermined introduction pressure. And an on-off valve 3 for opening and closing the pressurized air introduction path 2, and a pressurized helium introduction path 4 for introducing helium (He) gas pressurized from a pressure vessel (not shown) into the culture chamber 1, An open / close valve 5 for opening and closing the pressurized helium introduction path 4 is provided.
[0026]
The pressurized air introduction path 2 and the pressurized helium introduction path 4 are respectively provided with flow rate adjustment valves 6 and 7 each consisting of a needle valve, and the pressurized air introduction path 2 is provided downstream of the flow rate adjustment valves 6 and 7. And the pressurized helium introduction path 4 are merged with the merge introduction path 8. In addition, bacteria are prevented from entering the culture chamber 1 by interposing a sterilization filter 9 in the merging introduction path 8.
[0027]
The culture chamber 1 is provided with an opening (not shown) and a lid for opening and closing the culture chamber 1. After the culture sample 10 is put into the culture chamber 1 from the opening, the opening is sealed with the lid.
[0028]
The culture chamber 1 is connected to an atmosphere communication path 11 communicated with the atmosphere. The atmosphere communication path 11 is connected to a flow rate adjusting valve 12 and an on-off valve 13 including a needle valve, and another sterilization filter 9. Is interposed.
[0029]
Furthermore, a vacuum pump 14 and a bellows 15 are connected in parallel to the culture chamber 1, and a flow rate adjusting valve 17, which is a needle valve, and an open / close valve are connected to the suction path 16 connecting the vacuum pump 14 and the culture chamber 1. 18 is interposed.
[0030]
One end of the bellows 15 is closed by a fixed end plate 19 fixedly installed, and a movable end plate 20 that closes the other end is advanced and retracted toward the fixed end plate 19 by a pressurizing / depressurizing means 21.
[0031]
The pressurizing / depressurizing means 21 includes a motor 22, a crank arm 23 that is rotationally driven by the motor 22, and a connecting rod 24 that connects the movable end plate 19 of the bellows 15 to the crank arm 23. The distance between the connecting contact 25 connected to the crank arm 23 and the rotational axis of the crank arm 23 can be adjusted.
[0032]
A heater 26 is provided on the lower side of the culture chamber 1, referring to the temperature detected by the temperature sensor 27 provided in the culture chamber 1 so that the temperature in the culture chamber 1 becomes a predetermined culture temperature. A temperature adjusting device 28 for adjusting the operation of the heater 26 is provided.
[0033]
The internal pressure of the culture chamber 1 is detected by the pressure control unit 29 via the pressure monitor 29, and the pressure control unit 30 controls the open / close valves 3 and 5 so that the internal pressure of the culture chamber 1 becomes constant. It is.
[0034]
After the sample 10 is accommodated in the culture chamber 1 of this tissue cell pressure sensitivity test apparatus and the culture chamber 1 is sealed, the heater 26 is operated by the temperature adjustment device 28 to change the temperature in the culture chamber 1 to, for example, 0 ° C. After adjusting to a predetermined value of 50 ° C., the inside of the culture chamber 1 is purged with air by opening the opening / closing valve 3 of the pressurized air introduction path 2 and the opening / closing valve 13 of the atmosphere communication path 11.
[0035]
After that, the on-off valve 13 of the pressurized air introduction path 2 or the on-off valve 5 of the pressurized helium introduction path 4 is opened after closing the on-off valve 13 of the atmosphere communication path 11 to increase the pressure in the culture chamber 1. Is maintained at a predetermined introduction pressure, and the on-off valve 13 of the atmospheric communication passage 11 is opened after a predetermined time has elapsed, so that the culture sample 10 is applied at a predetermined pressure for a predetermined time as shown in FIG. Pressure (constant pressure test).
[0036]
Further, as described above, the inside of the culture chamber 1 is purged with air, and the open / close valve 13 of the atmospheric communication passage 11 is closed, and then the open / close valve 3 of the pressurized air introduction passage 2 or the open / close valve 5 of the pressurized helium introduction passage 4. To maintain the pressure in the culture chamber 1 at a predetermined introduction pressure for a predetermined time, and to open the on-off valve 13 of the atmosphere communication path 11 for a predetermined time to bring the internal pressure of the culture chamber 1 to atmospheric pressure. By alternately repeating the depressurization, as shown in FIG. 2 (B), the internal pressure of the culture chamber 1 can be changed to a rectangular pulse at atmospheric pressure or higher, and the culture sample 10 is rectangular at atmospheric pressure or higher. Pressurization can be performed at a pressure that changes in a pulse shape (rectangular pulse pressurization test).
[0037]
Further, the inside of the culture chamber 1 is purged with air, and the open / close valve 13 of the atmospheric air passage 11 is closed, and then the open / close valve 3 of the pressurized air introduction path 2 or the open / close valve 5 of the pressurized helium introduction path 4 is opened. After raising the pressure in the culture chamber 1 to a predetermined upper limit pressure, the pressurizing / depressurizing means 21 is operated to repeatedly expand and contract the space in the bellows 15, as shown in FIG. By changing the internal pressure of the culture chamber 1 to a triangular wave shape, the culture sample 10 can be pressurized with a pressure that changes in a triangular pulse shape above atmospheric pressure (triangular pulse pressure test).
[0038]
On the other hand, after purging the inside of the culture chamber 1 and closing the opening / closing valve 13 of the atmosphere communication path 11 as described above, the vacuum pump 14 is operated for a predetermined time and then the opening / closing valve 13 of the atmosphere communication path 11 is opened. By performing the valve operation, a predetermined negative pressure can be applied to the culture sample 10 for a predetermined time as shown in FIG.
[0039]
Furthermore, by to repeat the opening of the opening and closing valve 13 of the hydraulic and air communication passage 11 of the vacuum pump 14 periodically, as shown in FIG. 2 (E), is changed into a rectangular pulse form below atmospheric pressure The negative pressure which changes to a rectangular pulse shape below atmospheric pressure can be applied to the culture sample 10 (rectangular pulse decompression test).
[0040]
Furthermore, after purging the inside of the culture chamber 1 as described above and closing the open / close valve 13 of the air communication passage 11, the vacuum pump 14 is operated to reduce the internal pressure of the culture chamber 1 to a predetermined negative pressure. Then, by operating the pressurizing / depressurizing means 21 and repeatedly expanding and reducing the space in the bellows 15, the internal pressure of the culture chamber 1 is changed to a triangular wave shape as shown in FIG. Can be pressurized with a pressure that changes in a triangular pulse shape below atmospheric pressure (triangular pulse decompression test).
[0041]
Thus, when changing the internal pressure of the culture chamber 1 by expansion / contraction of the space in the bellows 15, the upper limit of the internal pressure of the culture chamber 1 is made higher than the atmospheric pressure by adjusting the expansion / contraction state of the bellows 15, The lower limit can be made lower than the atmospheric pressure, the internal pressure of the culture chamber 1 is changed alternately above and below the atmospheric pressure, and a pressure sensitivity test (alternating pressure fluctuation test) of tissue cells against this alternately changing pressure is performed. It can be carried out.
[0042]
That is, for example, the inside of the culture chamber 1 is purged with air, and the bellows 15 is reduced so that the space in the bellows 15 is minimized, and the on-off valve 13 of the atmosphere communication path 11 is closed to introduce pressurized air. When the opening / closing valve 3 of the passage 2 or the opening / closing valve 5 of the pressurized helium introduction passage 4 is opened to increase the pressure in the culture chamber 1 to a predetermined upper limit pressure and then the bellows 15 is enlarged, FIG. As shown, the internal pressure of the culture chamber 1 can be reduced to below atmospheric pressure, and the bellows 15 are continuously expanded and contracted to reduce the pressure in the culture chamber 1 to an upper limit higher than atmospheric pressure and lower than atmospheric pressure. Can be changed to a triangular pulse shape between the lower limit and the bellows 15 can be changed into a trapezoidal or rectangular pulse shape as shown in FIG.
[0043]
Note that the rectangular pulse wave pressurization test or the rectangular pulse wave decompression test described above can be applied to a pressure change test of about 10 cycles per minute. However, when the expansion / contraction of the bellows 15 is used, the rotation speed of the motor 19 is changed. By controlling the frequency of the pulse, it is possible to arbitrarily change the frequency of the pulse. For example, a very high cycle pressure change test of about 600 times per minute can be performed.
[0044]
In addition, a safety valve that allows the culture chamber 1 to communicate with the atmosphere when the internal pressure of the culture chamber 1 exceeds a certain level, or a safety valve that allows the culture chamber 1 to communicate with the atmosphere when the internal pressure reaches a certain level or less. May be provided.
[0045]
Furthermore, a cooler may be provided together with or in place of the heater 23 to cool the culture sample 10 in the culture chamber 1 to a temperature lower than room temperature.
[0046]
Furthermore, in this embodiment, a bellows 15 is provided separately from the culture chamber 1 so as to expand and contract the interior space. For example, the culture chamber 1 is slidable inside the cylinder and the inside thereof and is airtight. It is also possible to configure such that the volume of the culture chamber 1 can be expanded or contracted by driving the piston with the pressurizing / depressurizing means 21.
[0047]
In addition, the form of changing the internal pressure of the culture chamber 1 is not limited to a rectangular waveform or a triangular waveform, and can be changed to, for example, a sine curve shape or a step shape.
[0048]
In addition, in the above embodiment, air and helium are used as the pressurized gas, but in addition, carbon dioxide (CO ₂ ), oxygen (O ₂ ), hydrogen (H ₂ ), etc. are used as the pressurized gas. Can be used.
[0049]
【The invention's effect】
As described above, according to the first invention, in addition to the conventional pressurization test in which a constant pressure is applied to the sample, a rectangular pulse pressurization test in which a pressure fluctuating above atmospheric pressure is applied, a triangular pulse Pressure fluctuation such as pressure fluctuation test such as pressure test, decompression test applying negative pressure below atmospheric pressure, rectangular pulse wave decompression test applying negative pressure fluctuating below atmospheric pressure, triangular pulse wave decompression test Various tests such as a decompression test and an alternating fluctuating pressure test in which a pressure varying between a pressure higher than atmospheric pressure and a pressure lower than atmospheric pressure is applied are possible. Therefore, the pressure sensitivity of tissue cells can be tested in various ways, and the effect of elucidating the pathological mechanism and studying the pharmacological effect can be greatly expanded.
[0052]
In addition, according to the second invention, the space for the volume of gas in the variable communicates with the culture chamber, and hand stage Ru increase the internal pressure of the culture chamber by reducing the volume of gas in this space, this space since expanding the volume of the gas and a means for Ru reduce the internal pressure of the culture chamber, after enclosing the sample into the culture chamber, Ri by to scaling the volume of gas in the space, the internal pressure of the culture chamber The cycle time for increasing / decreasing the flow rate can be made shorter than in the case of the first invention. For example, the internal pressure of the culture chamber can be increased / decreased at a high frequency of about 600 times per minute, elucidating the pathological mechanism and the pharmacological effect. There is an effect that the scope of research can be further expanded dramatically.
[0053]
In particular, according to the second invention, it is easy to maintain the sealing properties of the culture chamber and the space communicating therewith, and there is no risk of problems such as vibration of the culture chamber, and a more reliable test. The effect which can be performed is acquired.
[Brief description of the drawings]
FIG. 1 is a configuration diagram of the present invention.
FIG. 2 is a culture chamber internal pressure waveform diagram of various susceptibility tests using the present invention.
FIG. 3 is a configuration diagram of a conventional example.
FIG. 4 is a configuration diagram of another conventional example.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 Culture chamber 2 Pressurized air introduction path 3 On-off valve 4 Pressurized helium introduction path 5 On-off valve 8 Confluence introduction path 10 Culture sample 11 Atmospheric communication path 13 On-off valve 14 Vacuum pump 15 Bellow 16 Suction path 18 On-off valve 21 Pressure reducing means

Claims (2)

A volume-invariant culture chamber that contains and seals a sample of living tissue, a pressurizing means for introducing a gas into the culture chamber at a predetermined introduction pressure, and an atmosphere connected to the culture chamber via an opening / closing means And a vacuum pump that is connected to the culture chamber and depressurizes the inside of the culture chamber, and switches each connection state between the pressurizing means, the air communication path, and the vacuum pump and the culture chamber. A tissue cell pressure sensitivity test apparatus characterized in that the pressure applied to the sample is changed by changing the internal pressure.   A space for changing the volume of gas is communicated with the culture chamber, a pressurizing means for reducing the volume of gas in the space to increase the internal pressure of the culture chamber, and expanding the volume of gas in the space to increase the volume of the gas. The tissue cell pressure sensitivity test apparatus according to claim 1, further comprising a decompression unit that reduces the internal pressure of the culture chamber.

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