CN210012859U - Multi-channel differential traction device for research on in-vitro axon stress mechanical response mechanism - Google Patents

Abstract

The utility model provides a multi-channel differential pulling device for studying the stress response mechanism of axons in vitro, including a culture and pulling control system and a mechanical device, wherein the culture and pulling control system includes a cell incubator , upper computer, controller and drive motor; the mechanical device includes a first coupling, a gear shaft, a gear shaft seat, a second coupling, a first lead screw nut linear slide table, and a second lead screw nut linear slide A stage, a cell pulling and growing device, a device support frame, and a base; the lead screw rotation directions of the first lead screw nut linear slide platform and the second lead screw nut linear slide platform are different. The device can be adapted to realize axonal pulling growth experiments with multiple samples, and can provide flexible testing pulling speed.

Description

A multi-channel differential traction device for the study of axonal stress response mechanisms in vitro

technical field

The utility model relates to the field of biomedical experiments, in particular to a multi-channel differential pulling device used for studying the stress response mechanism of axons in vitro.

Background technique

With the development of the economic level of all countries in the world, the incidence of spinal cord injury is increasing year by year. In developed countries, the incidence of spinal cord injury is approximately 13.3-45.9 persons/million persons/year. In China, there are about 60,000 new spinal cord injury patients every year. The current clinical treatment of spinal cord injury mainly includes surgery, drug therapy and long-term exercise rehabilitation. However, the massive loss of neural tissue and the failure of regenerative function make current treatments very limited.

At present, the main clinical neuroreparation procedures are autologous nerve transplantation and cell transplantation, which can promote the regeneration, repair and remodeling of damaged or damaged nerves, reconstruct neuroanatomical projection pathways and circuits, regulate and improve nerve signal conduction, and finally realize nerve function. repair. However, the two methods have their own hidden dangers: the autologous nerves cannot meet the needs of repairing a large number of damaged nerves due to the limited source and small diameter, and the nerves that are cut off will permanently lose their function and may form neuromas. Risk; cell transplantation through the injury target route may cause damage to the local brain tissue and spinal cord tissue, and the injury is more serious when multi-target transplantation is performed. Transplantation through the blood route is greatly affected by blood components and metabolic factors in the body. Influence of the brain barrier, cells are transported through the cerebrospinal fluid, adhere to the surface of the pia mater at the injury site, and may infiltrate extensively into the brain tissue in addition to the possible infiltration of nerve roots.

Chinese invention patent 201410403385.6 discloses a nerve axon pulling and growing device, which consists of two parts, a culture and pulling control system and a mechanical device. The controller connects and drives the stepping motor to rotate, and drives the linear slide of the ball screw at one end of the coupling to generate displacement. Indirect traction on nerve axons. However, only two sets of pulling devices can be connected to this device, and the speed of the pulling blocks is the same, and the number of nerve samples is not sufficient, so the effect of pulling speed on the growth of nerve axons cannot be well tested. The pulling acceleration is large, and it is easy to cause damage to the nerve.

Utility model content

The purpose of the present utility model is to provide a nerve axon pulling and growing device that can be adapted to realize multiple samples, and can provide a flexible test pulling speed.

The technical scheme of the present invention is as follows.

A nerve axon pulling and growing device, comprising a culture and pulling control system and a mechanical device, wherein:

The culturing and pulling control system includes a cell incubator, an upper computer, a controller and a drive motor;

The mechanical device includes a first coupling, a gear shaft, a gear shaft seat, a second coupling, a first lead screw nut linear slide, a second lead screw nut linear slide, a cell pulling and growing device, and a device support The first lead screw nut linear slide table and the second lead screw nut linear slide table have different screw rotation directions.

Preferably, the cell culture incubator is provided with a hole communicating with the outside world, for connecting the controller through the data line of the driving motor.

Preferably, the hole is located on the rear side of the cell culture incubator, and the connection between the hole and the data cable is sealed with silica gel.

Preferably, the upper computer can control the parameters of the pulling process, including one or more of the displacement, speed, and duration of cell pulling.

Preferably, there are multiple gear shafts, corresponding to each of the first lead screw nut linear slide table and the second lead screw nut linear slide table; the first coupling connects the drive motor is connected with one of the gear shafts; the second coupling connects each gear shaft with the corresponding lead screw nut linear slide table.

Preferably, there are a plurality of the first lead screw nut linear slide table and the second lead screw nut linear slide platform, and they are arranged alternately, and the corresponding gear shafts of the two adjacent lead screw nut slide platforms are meshed with each other.

Preferably, each lead screw of the first lead screw nut linear slide table and the second lead screw nut linear slide platform has a different pitch, so that each nut can generate different displacements.

Preferably, the controller can receive an instruction from the host computer to drive the drive motor to rotate; the drive motor can drive a plurality of gear shafts to rotate, thereby driving the lead screw nut linear slide to generate linear displacement.

Preferably, there are a plurality of the cell pulling and growing devices, respectively corresponding to each of the first lead screw nut linear slide table and the second lead screw nut linear slide table; the cell pulling growth device is provided in On the device support frame, together with the first and second lead screw nut linear slide tables, and the gear shaft seat are fixed on the base, and the base is placed in the cell culture incubator;

Each of the cell pulling growth devices includes a culture seat, a cover, an integrated pulling member, a pulling membrane and a bottom membrane; the integrated pulling block and the rod can be driven by the nut of the lead screw nut linear slide table. Movement of the stretched membrane, thereby pulling the nerve axon.

Preferably, the culture base is an integrated rectangular structure; the integrated pulling member includes a pulling block part and a pulling rod part; the pulling block part is located in the middle of the culture base, and the pulling rod part is located in the culture base the center of the end of the base; the bottom film is adhered to the bottom groove of the culture base; the cover is placed on the grooves on both sides and the upper surface of the culture base and fixed.

The advantages of the present utility model are:

(1) The utility model constructs a multi-channel, differential nerve axon pulling device, which can simultaneously control a plurality of independent pulling devices under the same culture environment, and realizes the length of the axon growth in the arithmetic progression, The speed is convenient for experimental comparison, which can not only verify the feasibility of mechanical pulling to stimulate nerve growth, but also test the effect of pulling speed on the growth of nerve axons.

(2) The utility model can collect the tensile stress data through the force sensor, analyze the influence of the stress on the growth of the nerve axon, and create the optimal condition for culturing the nerve tissue with perfect function in vitro, which is used for the repair of nerve damage, and in order to realize the In vivo implantation of a traction device for nerve repair sets the technical tone.

Description of drawings

1 is a schematic structural diagram of a nerve axon pulling and growing device of the present invention;

Fig. 2 is the mechanical structure diagram of the cell pulling growth device in the utility model;

Fig. 3 is the top plan view without cover of the cell pulling growth device in the utility model;

4 is a side view of an integrated pulling block and a rod in the utility model;

Fig. 5 is the pulling principle diagram of the cell pulling growth device in the utility model;

FIG. 6 is a structural diagram of axon pulling growth control in the present invention.

Detailed ways

The technical solutions in the embodiments of the present utility model will be described in detail and clearly below with reference to the accompanying drawings in the embodiments of the present utility model. The specific embodiments described herein are only used to explain the present invention, and do not limit the present invention.

As shown in FIG. 1 , the present invention relates to a multi-channel, differential pulling device for studying the stress response mechanism of axons in vitro, which is mainly composed of two parts: a culture and pulling control system and a mechanical device. The culture and pulling control system includes a cell incubator 1, a host computer 2, a controller 3 and a DC motor 4, and the mechanical device includes a first coupling 5 connected to the DC motor, a gear shaft 6, and a second coupling connected to the gear shaft. device 7 , left-handed screw nut linear slide 8 , right-handed screw nut linear slide 9 , cell pulling and growing device 10 , device support frame 11 , base 12 .

The rear side of the cell culture incubator 1 is provided with a hole communicating with the outside world, the data line of the DC motor 4 is connected to the controller 3 through the hole, and the connection between the hole and the data line is sealed with silica gel. The cell culture incubator 1 is a sealed structure, which can provide the carbon dioxide, temperature and humidity environment required for cell growth. In a preferred embodiment, the cell culture incubator 1 is set to ensure the culture conditions of 37°C temperature, 95% relative humidity and 5% carbon dioxide, such an environment can provide the best carbon dioxide, temperature and humidity for cell growth. surroundings.

The upper computer 2 can control the parameters of the pulling process through programming, including the speed, displacement and duration of cell pulling. The controller 3 receives the control command of the host computer 2, thereby driving the DC motor 4 to rotate. The DC motor is used for smooth and continuous output. Each rotation of the stepping motor will generate excessive instantaneous acceleration, which is prone to excessive traction. The pulling force causes the axon to rupture.

There are a plurality of the left-handed screw nut linear slide table 8 and the right-handed screw nut linear slide table 9 respectively, and preferably have an equal number. There are a plurality of the first coupling 5 connected to the DC motor, the gear shaft 6, the second coupling 7 connected to the gear shaft, and the cell pulling and growing device 10, which are respectively connected with the left-hand screw nut linear slide table 8. , Each of the right-hand screw nut linear slide tables 9 corresponds.

The DC motor 4 drives the gear shaft 6 at one end of the first coupling 5 connected to the DC motor to rotate, so that each gear shaft 6 drives the left-handed screw nut at one end of the second coupling 7 connected to the gear shaft. The slide table 8 and the right-hand screw nut linear slide table 9 are displaced. Due to the transmission between the gear shafts, the steering of two adjacent gear shafts is different, so the lead screws with different rotation directions are designed. Because the pitch of each lead screw is different, the displacement of the nut is different, so that the nerve axon is indirectly pulled by the integrated pulling block fixed on the linear slide nut and the rod 14, and the displacement and speed of the pulling are Different.

In order to prevent the axonal rupture due to excessive mechanical pulling displacement, the pulling time is 4-5 minutes every day, and the pulling displacement is about 0.5 mm. Correspondingly, in a preferred embodiment, for the left-handed lead screw nut linear slide table 8 and/or the right-handed screw nut linear slide table 9, a lead screw with a pitch of 0.5 mm can be selected according to the regulation of the fine thread, and the motor The rotational speed is set to 1 revolution for 5 minutes. Such a lead screw can meet the requirements of axon pulling and mechanical pulling, and can also ensure that the overall mechanical device is easy to process and manufacture.

In a preferred embodiment, the pitch of each lead screw with different left-handed and right-handed directions is set as an arithmetic progression, so that the displacements of each nut are also an arithmetic progression. In a more preferred embodiment, the displacement of each nut is set to be 1 times, 2 times, 3 times, and 4 times of a minimum pitch, so that the corresponding cell pulling and growing device 10 pulls the same displacement and speed. show the corresponding relationship.

Among them, the left-handed screw nut linear slide 8 and the right-handed screw nut linear slide 9 have a through hole in the middle and a threaded hole at the top for fixing the nut, the integrated pulling block and the rod.

As shown in FIGS. 2 to 5 , the cell pulling and growing device 10 has a cuboid structure, consisting of a culture base 13 , an integrated pulling block and rod 14 , a support block 15 , a cover 16 , a bottom film 17 , and a pulling film 18 composition.

Wherein, the culturing seat 13 is an integrated design structure. The support block 15 is located on the left side of the culture seat 13, and both ends are fastened with screws. The bottom film 17 is adhered to the small groove at the bottom of the culture seat 13 by coating with silica gel for cell culture. For the convenience of observation, the cover 16 is made of transparent material and fastened with screws.

Wherein, the through hole in the middle of the support block 15 can not only support the integrated pulling block and the rod 14, but also realize the guiding function of its movement. The top of the support block 15 is provided with three screw through holes, the upper and lower two are used for its fixing, and the screw through hole in the middle is used to fix the integrated pulling block and the rod 14 to prevent the integrated pulling block and the rod 14 from being in the cell culture. Device is loose.

The left side of the integrated pulling block and the rod 14 is the pulling rod part, and the right side is the pulling block part. The pulling rod part passes through the middle through hole of the support block 15, extends to the outside of the culture base 13, passes through the nuts of the screw nut linear slide tables 8 and 9, and is fastened with screws to integrate the two. The T-shaped frame on the upper part of the pulling block is on the middle groove of the culture base 13, and the arc at the lower part of the pulling block is used for the pulling of the pulling film 18, so that the extension part of the pulling film is attached to the bottom film. 17.

Wherein, the pulling film 18 is a rectangular transparent film, which is vertically adhered to the integrated pulling block and the pulling block part of the rod 14 with silica gel. As shown in FIG. 5 , the extension part of the pulling membrane 18 is in close contact with the base membrane 17 , and the axons attached to the pulling membrane 18 and the base membrane 17 can be mechanically stimulated by mechanical pulling.

Wherein, the culture seat 13 and the support block 15 of the cell pulling and growing device 10 are made of polytetrafluoroethylene material, the cover 16 is made of plexiglass material, and the integrated pulling block and the rod 14 are made of stainless steel material.

In this embodiment, in order to stretch and grow nerve axons, a set of nerve axon pulling and growing devices is designed.

When using, the pulling film 18 and the bottom film 17 are assembled first. After the device is assembled, place it under an ultraviolet lamp for several hours until the silica gel solidifies, and then add sterile water to soak for several days until the release of the silica gel acetic acid is completed, and then the cell culture can be carried out. The nerve cells or tissues are placed on both sides of the pulling membrane 18 and the base membrane 17 within 100 microns of each other, and the axons can be mechanically pulled after the nerve cells on both sides form a synaptic connection.

In this example, as shown in FIG. 6 , the control command of the host computer 2 is downloaded to the controller 3 by the data cable, so as to drive the DC motor 4 to rotate, and then drive the gear shaft to rotate through the first coupling 5, and then through the second The coupling 7 drives the nut of the lead screw nut linear slide table 8 and 9 to generate displacement, and then drives the integrated pulling block and the rod 14 fixed with the nut to move, and finally the integrated pulling block and the rod 14 drive the pulling film 18 It moves on the basement membrane 17 , thereby pulling the nerve axons growing on the stretcher membrane 18 and the basement membrane 17 .

In this embodiment, the pulling film 18 and the bottom film 17 are made of polychlorotrifluoroethylene film, which has good biocompatibility, is transparent, and is resistant to high temperature and high pressure, is not easily deformed, and is convenient for sterilization, observation and pulling. Preferably, the pulling film 18 and the bottom film 19 are respectively 50 μm and 198 μm polychlorotrifluoroethylene films.

In this embodiment, four independent cell pulling growth devices 10 are pulled simultaneously, so that the pulling speeds and displacements of the four independent cell pulling growth devices 10 are different. Further, according to the experimental requirements, the number of cell pulling growth devices 10 can be increased, and the number of gear shafts 6, the second coupling 7, the left-handed screw nut linear slide 8, and the right-handed screw nut linear slide 9 can be increased at the same time. number, to achieve a more diverse pulling speed and displacement of the cell pulling growth device 10 .

Those skilled in the art can understand that although there are four groups of cell pulling and growing devices and lead screw nut linear slide tables in this embodiment, the present invention is not limited to this. The number of the cell pulling growth device and the lead screw nut linear slide table can be set according to the actual number of samples required. In a preferred embodiment, the left-handed lead screw nut linear slide table and the right-handed screw nut linear slide table are set as The same number; in a more preferred embodiment, the number of the cell pulling growth devices is set to a multiple of 4.

In an optional embodiment (not shown in the figure), electrode contacts can be plated on the pulling membrane 18 and the base membrane 17, and are connected with a multi-channel nerve signal recording and stimulation system for recording nerve signals, and Selective stimulation of different sites of the nerve. A thin film force sensor can be attached to the pulling film 18 for recording the pulling stress.

In another optional embodiment, a set of electrical stimulation scheme of nerve axons is provided, and the base film 17 and the pulling film 18 are a flexible circuit board with a transparent microelectrode array. After the nerve axons are stretched and grown, the stretched axons can be electrically stimulated, and the corresponding nerve electrophysiological signals can be recorded.

Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present invention.

Claims (10)

1. A multi-channel differential pulling device for studying the stress response mechanism of axons in vitro, comprising a culture and pulling control system and a mechanical device, wherein: The culturing and pulling control system includes a cell incubator, an upper computer, a controller and a drive motor; The mechanical device includes a first coupling, a gear shaft, a gear shaft seat, a second coupling, a first lead screw nut linear slide, a second lead screw nut linear slide, a cell pulling and growing device, and a device support rack, base; It is characterized in that the lead screw rotation directions of the first lead screw nut linear slide table and the second lead screw nut linear slide platform are different. 2. The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to claim 1, wherein the cell culture incubator is provided with a channel that communicates with the outside world, and is used for passing through all the channels. The data line of the drive motor is connected to the controller. 3 . The multi-channel differential pulling device for in vitro axonal stress response mechanism research according to claim 2 , wherein the hole is located on the rear side of the cell incubator, and the hole is connected to a data cable. 4 . The connection is sealed with silicone. 4. The multi-channel differential pulling device for in vitro axonal stress response mechanism research according to claim 1, wherein the upper computer can control the parameters of the pulling process, including the displacement of cell pulling One or more of , Speed, Duration. 5 . The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to claim 1 , wherein there are multiple gear shafts, which are respectively in line with the first lead screw nut. 6 . The sliding table corresponds to each of the second lead screw nut linear sliding tables; the first coupling connects the drive motor with one of the gear shafts; the second coupling connects each gear The shaft is connected with the corresponding lead screw nut linear slide. 6. The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to claim 5, wherein the first lead screw nut linear slide table and the second lead screw nut linear slide There are a plurality of tables respectively, and they are arranged alternately, and the corresponding gear shafts of the two adjacent lead screw nut sliding tables are meshed with each other. 7 . The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to claim 6 , wherein the first lead screw nut linear slide table and the second lead screw nut linear slide. 8 . Each lead screw of the table has a different pitch, whereby each nut can produce a different displacement. 8 . The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to claim 5 , wherein the controller can receive an instruction from the upper computer to drive the The driving motor rotates; the driving motor can drive a plurality of gear shafts to rotate, thereby driving the linear sliding table of the lead screw nut to generate linear displacement. 9. The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to any one of claims 1-8, wherein the cell pulling and growing devices are multiple, respectively corresponding to each of the first lead screw nut linear slide table and the second lead screw nut linear slide platform; the cell pulling and growing device is arranged on the device support frame, together with the first and second The lead screw nut linear slide table and the gear shaft seat are fixed on the base, and the base is placed in the cell incubator; Each of the cell pulling growth devices includes a culture seat, a cover, an integrated pulling member, a pulling membrane and a bottom membrane; the integrated pulling block and the rod can be driven by the nut of the lead screw nut linear slide table. Movement of the stretched membrane, thereby pulling the nerve axon. 10 . The multi-channel differential pulling device for studying the stress response mechanism of axons in vitro according to claim 9 , wherein the culture base is an integrated rectangular structure; the integrated pulling member comprises: 11 . a pulling block part and a pulling rod part; the pulling block part is located in the middle of the culture base, and the pulling rod part is located at the center of the end of the culture base; the bottom film is adhered to the bottom groove of the culture base; The cover is placed on the grooves on both sides and the upper surface of the culture seat and fixed.

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