Method for preparing biological tissue for transplantation

Abstract

A method for processing biological tissue for transplantation, comprising the steps of extracting the tissue, pre-cleaning, placing the tissue and a processing solution together in a packaging bag, sealing the packaging bag, and then placing the packaging bag in a fluid medium and applying water purification pressure to the packaging bag, the high-pressure treatment being performed in which the components of the processing solution include a nucleic acid degrading enzyme and a buffer solution, and post-cleaning in which the tissue after the high-pressure treatment is washed.

Description

[Technical field]

[0001] The present application relates to the field of tissue transplantation, and more specifically to methods for preparing biological tissue for transplantation. [Background technology]

[0002] The tissue transplantation process is the transplantation of autologous tissue or artificial materials into a specific area or part of the body to repair degeneration, deformation, or tissue defects caused by congenital or acquired factors. Tissue transplants usually include bone and cartilage transplants, skin and fat transplants, mucosa transplants, fascia transplants, muscle transplants, nerve transplants, composite tissue transplants, biomaterial filling, and tissue engineered tissue transplants.

[0003] Compared with autotransplantation and allotransplantation, which use healthy tissue from oneself or another person, transplantation of artificial materials such as artificial objects and bio-derived materials is gradually becoming a popular option for tissue transplantation, but the body usually tends to reject artificial materials. In order to solve the above problems, decellularization technology, which is a description of processing bio-derived materials, has been attracting attention in recent years.

[0004] Decellularization techniques form biological tissue scaffolds that are non- or lowly immunogenic by removing cells from allogeneic or xenogeneic tissues by chemical and physical methods. Methods for decellularizing biological tissues include the use of surfactants, enzymes, acidifiers, and ultra-high pressure hydrostatic pressure treatment. Decellularized tissues after ultra-high pressure hydrostatic pressure treatment, which does not use toxic chemicals, have the advantage of being non-cytotoxic compared to chemical methods, and the remaining active substances have a superior effect of inducing host cell differentiation after transplantation compared to chemical methods.

[0005] However, the inventors found that when treating cell tissues with ultra-high hydrostatic pressure, it is necessary to control the temperature and pressure, but the properties of the decellularized tissue finally obtained under the same temperature and pressure treatment conditions may not be consistent. This indicates that ultra-high hydrostatic pressure may cause instability in the decellularization process of cell tissues, which is a very serious problem in the clinical application of decellularized tissues. Summary of the Invention [Problem to be solved by the invention]

[0006] The ultra-high hydrostatic pressure improves the stability of the cell tissue treatment, thereby obtaining decellularized tissue with low cytotoxicity and excellent cell differentiation induction effect.The present application provides a method for treating biological tissue for transplantation. [Means for solving the problem]

[0007] The method for treating biological tissue for transplantation provided in this application employs the following technical solutions. A method of preparing biological tissue for transplantation, comprising the steps of: harvesting tissue; A pre-cleaning step; The high-pressure treatment involves placing the tissue and a treatment solution together in a packaging bag, sealing the bag, placing the bag in a fluid medium, and applying water purification pressure to the packaging bag, and includes high-pressure treatment in which the components of the treatment solution include a nucleic acid degrading enzyme and a buffer solution, and a post-washing step in which the tissue after the high-pressure treatment is washed.

[0008] By adopting the above technical solution, the process of obtaining decellularized tissue using ultra-high hydrostatic pressure needs to be carried out in a fluid medium, so in the research process, the inventor first changed the type of fluid medium to improve the stability of biological tissue processing. The inventor processed biological tissue with ultra-high hydrostatic pressure using water, saline, a mixture of saline and glycerin, glucose, etc. as fluid media, but still did not obtain ideal results. The inventor speculated that the cellular components remaining in the biological tissue may affect the decellularization process.

[0009] Therefore, the present inventors considered that nucleases could be used to decompose nucleic acid components in biological tissues, and directly added nucleases to a fluid medium, and decellularized biological tissues by the cooperation of nucleases and ultra-high hydrostatic pressure. As a result, the present inventors unexpectedly discovered that the addition of nucleases improved the efficiency of removing cellular components in biological tissues. However, the stability of the decellularization process has not yet been sufficiently improved.

[0010] The inventor conducted many experiments and studies, but could not find a way to improve the stability of the decellularization process. Finally, the inventor thought that the process of obtaining decellularized tissue using ultra-high hydrostatic pressure needs to be carried out in a fluid medium, and intuitively thought of changing the type of fluid medium, but devised a way to solve the problem from a different angle. The inventor used a packaging bag to pre-package the biological tissue, and put a mixture of nuclease and buffer solution into the packaging bag as a treatment solution, and relatively isolated the biological tissue and the treatment solution from the large environment of the ultra-high hydrostatic pressure treatment. Finally, the inventor found that this operation can fully improve the stability of ultra-high hydrostatic pressure in the decellularization process of the cell tissue, and the treatment solution mixed with nuclease and buffer solution can be put into the packaging bag together with the biological tissue, and the treatment efficiency of the biological tissue can also be greatly improved, and the obtained decellularized tissue has low cytotoxicity and excellent cell induction differentiation effect.

[0011] Preferably, the buffer solution includes at least one of a PBS buffer solution and a HEPES buffer solution.

[0012] Preferably, in the high pressure treatment, the packaging bag has at least two layers, the tissue and the treatment liquid are located in the innermost packaging bag, and the remaining packaging bags are filled with the liquid medium.

[0013] By adopting the above technical solutions and using at least two layers of packaging bags, the performance of the final decellularized tissue is further improved.

[0014] Preferably, in the high pressure treatment, the packaging bag is two-layered, the tissue and the treatment liquid are located in the innermost packaging bag, and the outer packaging bag is filled with a liquid medium, and the density of the treatment liquid is greater than the density of the liquid medium.

[0015] By adopting the above technical solutions, the inventors found that the decellularization process is more stable when the density of the treatment liquid is greater than the density of the liquid medium, and the inventors deduced that this is because, in the process of pressure being transmitted to biological cell tissues, the density of the treatment liquid is greater, which can reduce the impact of the large environmental pressure of ultra-high purified water pressure on the small environmental pressure of the packaging bag, allowing pressure to be applied to the biological tissues more stably and uniformly.

[0016] Preferably, in the high-pressure treatment step, the packaging bag is placed in a high-voltage device to perform high-pressure treatment, the high-voltage device includes a base, a pressurizing chamber, and a water injection assembly, the pressurizing chamber includes a fixed chamber and a moving chamber, the fixed chamber is fixedly connected to the base, the inner wall of the fixed chamber is provided with a peripheral wall and opens at one axial end, the outer periphery of the moving chamber is provided with a peripheral wall and opens at one axial end, the open end of the moving chamber is inserted into the fixed chamber, the outer periphery wall of the moving chamber and the inner periphery wall of the fixed chamber are bonded together, a first seal is provided at the open end of the fixed chamber, and the first seal and the outer periphery wall of the moving chamber are abutted against each other, the base is provided with a drive assembly for driving the sliding movement or rotation on the spot of the moving chamber, a guide sleeve is fixedly connected to the inner wall of the fixed chamber, and a carrier cover is provided within the guide sleeve. the guide sleeve and the carrier cover are slidably mounted, and both of the guide sleeve and the carrier cover are provided with water-passing grooves passing therethrough, the outer peripheral wall of the carrier cover is provided with a spiral arc groove and a linear arc groove is further formed on the outer peripheral wall of the carrier cover, the linear arc groove passes through the carrier cover in the axial direction of the carrier cover, and the linear arc groove and the spiral arc groove are provided through each other, one end of the carrier cover extending from the guide sleeve is located in the moving chamber, a guide ball is rotatably connected to an inner wall of the moving chamber, the guide ball and the linear arc groove are provided in one-to-one correspondence, one end of the guide ball remote from the moving chamber is inserted into the linear arc groove or the spiral arc groove, and a spherical wall of the guide ball abuts against a groove wall of the linear arc groove or a groove wall of the spiral arc groove, and the water-injection assembly and the fixed chamber are provided through each other.

[0017] By adopting the above technical solution, the fixed chamber and the moving chamber are provided separately, so that when the moving chamber is separated from the fixed chamber and the carrier cover is exposed outside the fixed chamber, the tissue product can be easily stored and taken out. The driving assembly allows the moving chamber to slide in the axial direction, and the guide ball rolls in the linear arc groove, which realizes the moving chamber to slide in the axial direction on the fixed chamber, and the pressurizing chamber can be easily tightened. And when the driving assembly drives the carrier cover to rotate on the spot, the guide ball enters the spiral arc groove accordingly, so that a movement similar to thread feeding occurs between the carrier cover and the moving chamber, and the carrier cover can slide on the guide sleeve, and the carrier cover enters and exits the guide sleeve and then enters and exits the fixed chamber, so that the guide ball, the linear arc groove and the spiral arc groove of the present application cooperate with each other, and the carrier cover and the moving chamber cooperate with each other to realize the sliding movement of the moving chamber in the axial direction and at the same time the sliding movement of the carrier cover, which makes it easy to store the moving chamber and the carrier cover, and the operator can easily take out and store the product.

[0018] Preferably, both the inner peripheral wall of the guide sleeve and the outer peripheral wall of the carrier cover are provided with a magnetic layer.

[0019] By adopting the above technical solution, the friction force between the guide sleeve and the carrier cover can be increased, friction positioning can be performed on the carrier cover, and the free movement of the carrier cover can be reduced.

[0020] Preferably, the drive assembly includes a threaded shaft, a drive ring seat, a compression block, and a feed screw block, the threaded shaft is fixedly connected to one end of the moving chamber extending from the fixed chamber, the drive ring seat is rotatably connected to the base, and the base is provided with a drive part for driving the rotation of the drive ring seat, the one end of the threaded shaft away from the moving chamber is connected to the inner ring wall of the drive ring seat through the inner ring wall of the drive ring seat, there is a gap between the outer peripheral wall of the threaded shaft and the inner ring wall of the drive ring seat, at least three feed screw blocks are connected to the inner ring wall of the drive ring seat so as to be slidable in the radial direction of the drive ring seat, at least three of the feed screw blocks are uniformly distributed in the circumferential direction of the drive ring seat, and the feed screw block is connected to the inner ring wall of the drive ring seat so as to be slidable in the radial direction of the drive ring seat, a feed screw block abuts against the threaded shaft at one end thereof and is screw-connected to the threaded shaft; at least two of the compression blocks are connected to an inner ring wall of the drive ring seat so as to be slidable in a radial direction of the drive ring seat; a gap is provided between the compression block and the threaded shaft at one end thereof; at least three of the compression blocks are uniformly distributed in a circumferential direction of the drive ring seat; the at least three compression blocks are opposite to the sliding movement direction of the at least three feed screw blocks; a power assembly is provided on the drive ring seat for driving the sliding movement of the feed screw block and the compression block; a guide block is provided on the base so as to be slidable in an axial direction of the threaded shaft; and an interlocking assembly is provided between the guide block and the threaded shaft.

[0021] By adopting the above technical solution, after the driving ring seat rotates, the first gear and the second gear are driven to rotate due to the meshing effect, and then the first thread insert and the second thread insert are rotated due to the direction changing action of the first bevel gear set and the second bevel gear set, so that yarn feeding is generated between the first thread insert and the first thread block, and yarn feeding is generated between the second thread insert and the second thread block. Therefore, by reversing the feed direction of the first screw block and the second screw block, the compression block and the feed screw block can be made to slide in the opposite direction, so that the compression block is in close contact with the threaded shaft, the guide block is separated from the threaded shaft, and the threaded rotating rod can be driven to rotate by the driving rotating seat, and the moving chamber rotates, the compression block is separated from the threaded shaft, the feed screw block is abutted against the threaded shaft and is engaged with the thread on the threaded shaft accordingly, and at the same time, the guide block and the threaded shaft are fixed relatively by the interlocking assembly, so that when the driving ring seat rotates, the feed screw block generates a thread feed between the threaded shaft, and thus the threaded shaft drives the moving chamber to slide in the axial direction, which is convenient and quick.

[0022] Preferably, the power assembly comprises a drive ring gear, a first gear, a second gear, a first screw block, a second screw block, a first screw insert and a second screw insert, an accommodating cavity is provided within the drive ring seat, the drive ring gear is rotatably connected to a cavity wall of the accommodating cavity, and a rotary drive part is provided on the cavity wall of the accommodating cavity for driving the rotation of the drive gear. The first gear and the second gear are both rotatably connected to the cavity wall of the accommodating cavity and both mesh with the drive ring gear, the first gear and the feed screw block are arranged in a one-to-one correspondence, the second gear and the compression block are arranged in a one-to-one correspondence, the first thread insert is connected to the first gear via a first bevel gear set, the first thread block is fixedly connected to one end of the feed screw block extending into the accommodating cavity, and the first thread block and an inner circumferential wall of the first thread insert are screwed together, the second thread insert is connected to the second gear via a second bevel gear set, the second thread block is fixedly connected to one end of the compression block extending into the accommodating cavity, and the second thread block and an inner circumferential wall of the second thread insert are screwed together.

[0023] Preferably, the water injection assembly includes a water supply tank, a water inlet pipe head, a water outlet pipe head, a connecting pipe, and a spray ball head. The fixed chamber has an installation groove on an outer wall away from the movable chamber. The fixed chamber has a first pipe trough and a second pipe trough on an inner wall opposite to the open end. The first pipe trough and the second pipe trough are both passed through the installation groove. The water inlet pipe head is slidably mounted on the groove wall of the installation groove. The base is provided with a sliding part for driving the sliding movement of the water inlet pipe head. One end of the water inlet pipe head located within the installation groove is sealed. A water supply pipe and a water pump are provided between one end of the water inlet pipe head extending from the installation groove and the water supply tank. A first control valve is provided at one end of the water pump close to the fixed chamber. One end of the first pipe trough that passes through the installation groove and one end of the second pipe trough that passes through the installation groove are provided. are arranged side by side in the sliding direction of the water inlet pipe head, a water passage is formed through the peripheral wall of the water inlet pipe head, the water outlet pipe head is attached to the groove wall of the first pipe trough, the communicating pipe is provided on the groove wall of the second pipe trough, one end of the communicating pipe close to the water inlet pipe head and one end of the water outlet pipe head close to the water inlet pipe head are both provided with second seals, the groove wall of the water passage passes through the water outlet pipe head or through the communicating pipe, the spray ball head is fixedly connected to the inner wall facing the open end of the fixed chamber, a number of water passages are provided within the spray ball head, the water passages pass through a spherical wall of the spray ball head extending to the fixed chamber, one end of the communicating pipe remote from the water inlet pipe head is connected to the spray ball head and passes through all of the water passages, an exhaust pipe is connected to the top end of the fixed chamber, and a second control valve is attached to the exhaust pipe.

[0024] By adopting the above technical solution, when the water inlet passes through the water outlet pipe head, the water tank, the water pipe, and the water pump cooperate with the water inlet pipe head and the water outlet pipe head to normally inject the fluid medium into the pressurized chamber. After the high pressure treatment is completed, the water inlet pipe head is moved to pass the water inlet through the connecting pipe, so that the fluid medium or the washing water enters the spray ball head and is guided by the water passage, so that the fluid medium or the washing water is sprayed at multiple angles into the fixed chamber, and the fixed chamber and the moving chamber are cleaned to a certain extent, so that the cleanliness of the decellularized tissue treatment process can be improved. Effect of the Invention

[0025] In summary, the present application has the following beneficial effects: 1. In the present application, a mixture of nuclease and a buffer solution is used as the treatment solution, and then the treatment solution and biological tissue are packaged in advance and then placed in a fluid medium of a high-voltage device, the biological tissue and the treatment solution are relatively isolated from the large environment of the ultra-high hydrostatic pressure treatment, and the high-pressure treatment is performed on the biological tissue at a temperature of 20 to 25° C. The stability of the ultra-high hydrostatic pressure in the decellularization process of the cell tissue can be sufficiently improved, and the treatment efficiency of the biological tissue can be greatly improved, and the obtained decellularized tissue has low cytotoxicity and excellent cell induction differentiation effect.

[0026] 2. In the present application, a buffer solution is used as a treatment solution in combination with a nuclease, and can be used for decellularization of various biological tissues.

[0027] 3. The present application provides a high voltage device that allows convenient and quick storage and removal of the product. [Brief description of the drawings]

[0028] [Figure 1] FIG. 1 is a schematic diagram of a high-voltage device according to a third embodiment of the present invention. [Diagram 2] FIG. 2 is a cross-sectional view taken along line AA in FIG. [Diagram 3]FIG. 3 is an exploded view showing a moving chamber structure in the third embodiment of the present application. [Figure 4] FIG. 4 is a schematic diagram showing a water injection assembly structure in Example 3 of the present application. [Diagram 5] FIG. 5 is a cross-sectional view taken along line BB in FIG. [Figure 6] FIG. 6 is an enlarged view of a portion C in FIG. [Figure 7] FIG. 7 is a schematic diagram showing a drive assembly structure in the third embodiment of the present application. [Figure 8] FIG. 8 is a cross-sectional view taken along line DD in FIG. [Figure 9] FIG. 9 is a schematic diagram showing a power assembly structure in the third embodiment of the present application. [Figure 10] FIG. 10 is a schematic diagram showing an interlocking assembly structure in the third embodiment of the present application. [Figure 11] FIG. 11 is a cross-sectional view taken along line EE in FIG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] The present application will now be described in more detail with reference to examples.

[0030] Working Example Example 1 In this Example 1, a method for preparing biological tissue for transplantation is provided.

[0031] A method for preparing biological tissue for transplantation, comprising the steps of: Step 1: Extract the tissue Cut the carotid artery of a veterinary pig into 1 cm long segments at 4 °C.

[0032] Step 2: Pre-wash Physiological saline is used as a washing solution, and the excised porcine heart tissue is placed in the washing solution and oscillated at 25°C for 6 hours to complete the washing.

[0033] Step 3: High pressure treatment The washed vascular tissue and the treatment liquid are put together into a first packaging bag, and then the packaging bag is clad and sealed. The packaging bag containing the vascular tissue is then put into a second packaging bag, and at the same time, a liquid medium is added into the second packaging bag, and then the second packaging bag is clad and sealed. In this embodiment, there are no bubbles between the two layers of packaging bags.

[0034] Finally, the wrapped two-layered packaging bag is placed into a high-pressure treatment device, which is filled with a fluid medium. At a temperature of 20°C, a hydrostatic pressure of 980 MPa is applied to the packaging bag for 10 minutes to complete the decellularization process.

[0035] Here, the treatment liquid is a mixture of nuclease and HEPES buffer solution, and the mass ratio of the nuclease to the HEPES buffer solution is 1: 7. The liquid medium is water.

[0036] Step 4: After washing, The vascular tissue after high pressure treatment is placed in a cleaning solution and washed with oscillation at 4°C for 4 days, then washed with 80 wt% ethanol aqueous solution with oscillation for 3 days, and further washed with HEPES buffer solution with oscillation for 2 weeks to obtain decellularized tissue. The cleaning solution is the same substance as the treatment solution in this embodiment.

[0037] Example 2 In this Example 1, a method for preparing biological tissue for transplantation is provided.

[0038] A method for preparing biological tissue for transplantation, comprising the steps of: Step 1: Extract the tissue The bone marrow from a culinary pig is placed in a small block measuring 1cm x 1cm and kept at 4℃.

[0039] Step 2: Pre-wash Physiological saline is used as a washing solution, and the excised porcine heart tissue is placed in the washing solution and oscillated at 25°C for 6 hours to complete the washing.

[0040] Step 3: High pressure treatment The washed pig bone marrow tissue and the processing liquid are put together into a first packaging bag, and then the packaging bag is clad and sealed. Then, the packaging bag containing the pig bone marrow tissue is put into a second packaging bag, and at the same time, a liquid medium is added into the second packaging bag, and then the second packaging bag is clad and sealed. In this embodiment, there are no bubbles between the two layers of packaging bags.

[0041] Finally, the wrapped two-layered packaging bag is placed into a high-pressure treatment device, which is filled with a fluid medium. At a temperature of 20°C, a hydrostatic pressure of 980 MPa is applied to the packaging bag for 10 minutes to complete the decellularization process.

[0042] Here, the treatment liquid is a mixture of nuclease and PBS buffer solution, and the mass ratio of nuclease to HEPES buffer solution is 1:9.3, and the liquid medium is water.

[0043] Step 4: After washing, The porcine bone marrow tissue after high pressure treatment is placed in a washing solution and washed by oscillation for 2 weeks in an environment of 37° C., and then decellularized tissue is obtained. The washing solution is the same substance as the washing solution in this embodiment.

[0044] Example 3 In this embodiment, a high voltage device is provided.

[0045] 1 and 2, the high voltage equipment includes a base 1, a pressurized chamber 2, and a water injection assembly 3. In this embodiment, the pressurized chamber 2 includes a cylindrical fixed chamber 21 and a cylindrical moving chamber 22. One axial end of the fixed chamber 21 and one axial end of the moving chamber 22 are both open. A first base block 11 is provided on the base 1. The fixed chamber 21 is fixedly connected to the top end of the first base block 11 and is horizontal in the axial direction. The open end of the moving chamber 22 is inserted into the fixed chamber 21, and the outer peripheral wall of the moving chamber 22 and the inner peripheral wall of the fixed chamber 21 are bonded together. The moving chamber 22 may be slidable in the fixed chamber 21 in the axial direction, or may be rotatable in its own circumferential direction. A first seal 23 is provided at the open end of the fixed chamber 21. In this embodiment, the first seal 23 is a seal ring, and the inner wall of the ring of the first seal 23 is in contact with the outer circumferential wall of the moving chamber 22. A drive assembly 4 is provided on the base 1 for driving the rotation or sliding movement of the moving chamber 22. A water injection assembly 3 communicates with the fixed chamber 21.

[0046] 2 and 3, in this embodiment, a first link 24 is fixedly connected to one side of the inner peripheral wall of the fixed chamber 21 that is away from the open end, and a circular guide sleeve 25 is fixedly connected to the first link 24. The guide sleeve 25 and the fixed chamber 21 share a central axis, and a circular carrier cover 26 is inserted into the guide sleeve 25. A magnetic layer 27 is applied to both the inner peripheral wall of the guide sleeve 25 and the outer peripheral wall of the carrier cover 26. The carrier cover 26 slides on the inner peripheral wall of the guide sleeve 25 in the axial direction of the guide sleeve 25, and both the guide sleeve 25 and the carrier cover 26 are provided with water passage grooves 28. A spiral arc groove 261 is opened on the outer peripheral wall of the carrier cover 26, and two straight arc grooves 262 are further opened on the outer peripheral wall of the carrier cover 26, the two straight arc grooves 262 penetrate the carrier cover 26 in the axial direction of the carrier cover 26 and are evenly distributed in the circumferential direction of the carrier cover 26, and at the same time, the straight arc groove 262 is arranged through the spiral arc groove 261, and the widths and radians of the straight arc groove 262 and the spiral arc groove 261 are all the same and transition smoothly.

[0047] As shown in Figures 2 and 3, one end of the carrier cover 26 extending from the guide sleeve 25 is positioned within the movable chamber 22, a ring set 291 is fixedly connected to the inner wall of the movable chamber 22 via the second base block 221, the ring set 291 and the straight arc groove 262 are arranged in one-to-one correspondence, a guide ball 29 is rotatably connected within the ring set 291, the ring peripheral wall of the ring set 291 and the spherical wall of the guide ball 29 are in contact with each other, one end of the guide ball 29 away from the second base block 221 extends from the ring set 291, and one end of the guide ball 29 extending from the ring set 291 is inserted into the straight arc groove 262, and the spherical wall of the guide ball 29 and the groove wall of the straight arc groove 262 are in contact with each other.

[0048] Initially, the moving chamber 22 is separated from the fixed chamber 21, and most of the carrier cover 26 extends from the guide sleeve 25 and is located in a fixed position, and the product is placed on the carrier cover 26, and then the drive assembly 4 is started, and the moving chamber 22 moves axially toward the fixed chamber 21, and the moving chamber 22 is inserted into the fixed chamber 21, and the guide ball 29 rolls in the straight arc groove 262. After the moving chamber 22 reaches a predetermined position, the driving assembly 4 drives the moving chamber 22 to rotate on the spot, at this time, the guide ball 29 enters the spiral arc groove 261, the spherical wall of the guide ball 29 competes with the groove wall of the spiral arc groove 261 and rolls in the spiral arc groove 261, so that the carrier cover 26 moves axially to the guide sleeve 25, and after the carrier cover 26 reaches a predetermined position, the guide ball 29 moves to the linear arc groove 262, at this time, the driving assembly 4 drives the moving chamber 22 to move axially, so that the moving chamber 22 reaches an appropriate position in the fixed chamber 21 and completes the product loading.

[0049] Then, the water injection assembly 3 is used to inject fluid medium into the fixed chamber 21 and the movable chamber 22 until there are no bubbles in the fixed chamber 21 and the movable chamber 22. Then, the pressurized chamber 2 is pressurized according to needs, and the pressurizing method is to maintain the volume in the pressurized chamber 2 as it is and continue to add fluid medium by the water injection assembly 3 to pressurize, or to start the driving assembly 4 to make the guide ball 29 continue to slide along the linear arc groove 262, so that the movable chamber 22 slides toward the fixed chamber 21, thereby reducing the volume of the pressurized chamber 2 and pressurizing it.

[0050] After the high pressure treatment is completed, the pressure in the pressurized chamber 2 is released, and the fluid medium in the pressurized chamber 2 is discharged. Then, the driving assembly 4 is started, and the moving chamber 22 reaches a predetermined position from the fixed chamber 21 in the axial direction, and then the moving chamber 22 is driven to rotate on the spot, so that the guide ball 29 rolls in the spiral arc groove 261, and thus the carrier cover 26 extends from the guide sleeve 25, and then the moving chamber 22 is driven to move in the axial direction, so that the guide ball 29 rolls in the linear arc groove 262, and thus the moving chamber 22 leaves the fixed chamber 21, and at this time, the product can be taken out from the carrier cover 26 exposed to the outside.

[0051] 4 and 5, the water injection assembly 3 includes a water supply tank 31, a water inlet pipe head 32, a water outlet pipe head 33, a connecting pipe 34, and a spray ball head 35. A rectangular mounting groove 36 is provided on the outer wall of the fixed chamber 21 away from the movable chamber 22. The outer peripheral wall of the water inlet pipe head 32 is a rectangular tube and is slidably mounted on the groove wall of the mounting groove 36 in the axial direction of the fixed chamber 21. One end of the water inlet pipe head 32 is sealed within the mounting groove 36, and the outer peripheral wall of the water inlet pipe head 32 and the groove wall of the mounting groove 36 are in contact with each other. A third base block 37 is fixedly connected to the base 1, and a sliding part 38 is attached to the third base block 37. The sliding part 38 in this embodiment is a cylinder, and the driving end of the sliding part 38 is fixedly connected to the outer peripheral wall of one end of the water inlet pipe head 32 extending from the fixed chamber 21. A water supply pipe 39 is connected to one end of the water inlet pipe head 32 extending from the fixed chamber 21, and the water supply pipe 39 is connected to the water supply tank 31 via a water pump 391. A first control valve 392 is provided on the water supply pipe 39 at the end where the water pump 391 is close to the fixed chamber 21.

[0052] 5 and 6, a first pipe trough 361 and a second pipe trough 362 are arranged in the axial direction of the fixed chamber 21 on the upper wall of the mounting groove 36, and the first pipe trough 361 is located on one side of the second pipe trough 362 away from the moving chamber 22. The first pipe trough 361 and the second pipe trough 362 penetrate the inner wall facing the open end of the fixed chamber 21. The water outlet pipe head 33 is attached to the groove wall of the first pipe trough 361, and the water outlet pipe head 33 and the groove wall of the first pipe trough 361 are connected in a sealed manner and pass through the inside of the fixed chamber 21. The communicating pipe 34 is fixedly attached to the groove wall of the second pipe trough 362, the spray ball head 35 is fixedly connected to the inner wall facing the open end of the fixed chamber 21, a number of water passages 351 are opened in the spray ball head 35, the water passages 351 penetrate the spherical wall extending to the fixed chamber 21 of the spray ball head 35, one end of the communicating pipe 34 remote from the water inlet pipe head 32 is sealed connected to the spray ball head 35, and one end of the communicating pipe 34 remote from the water outlet pipe head 33 passes through all the water passages 351. In this embodiment, a second seal 322 is fixedly connected to one end of the communicating pipe 34 close to the water inlet pipe head 32 and one end of the water outlet pipe head 33 close to the water inlet pipe head 32. The second seal 322 is a seal ring. The two second seals 322 are always in contact with the upper wall of the water inlet pipe head 32, and a water port 321 is also provided through the upper wall of the water inlet pipe head 32. An exhaust pipe 211 is connected to the top end of the fixed chamber 21, and a second control valve 212 is attached to the exhaust pipe 211. A drain pipe 213 is connected to the bottom end of the fixed chamber 21, and a third control valve 214 is attached to the drain pipe 213.

[0053] At the beginning, the water port 321 is passed through the water outlet pipe head 33, the second seal 322 at the water outlet pipe head 33 is located on the circumferential side of the water port 321, the first control valve 392 and the second control valve 212 are opened, and at the same time, the second seal 322 at the communication pipe 34 is sealed against the water inlet pipe head 32 to seal the communication pipe 34. When the water pump 391 is started, the fluid medium is introduced into the fixed chamber 21, and after there are no bubbles in the pressurized chamber 2, the second control valve 212 and the first control valve 392 are closed, the water pump 391 is closed, the injection of the fluid medium is completed, and then the high pressure treatment is performed. After the high pressure treatment is completed, the third control valve 214 is opened, the fluid medium is discharged, and the movable chamber 22 is slid out of the fixed chamber 21 to take out the product. After the high pressure treatment is finished and the product is removed, the sliding part 38 is started to slide the water inlet pipe head 32 to the moving chamber 22, and the water port 321 is passed through the connecting pipe 34, and the second seal 322 in the connecting pipe 34 is sealed around the water port 321. Then, cleaning water is added to the water supply tank 31, and then the first control valve 392 is opened and the water pump 391 is started, so that the cleaning water enters the water passage 351 through the connecting pipe 34 and is further sprayed through the water passage 351 into the pressurized chamber 2 to clean the pressurized chamber 2 to a certain degree. In addition, the water injection assembly in this embodiment may also include a water tank and a water pipe connected between the water tank and the fixed chamber 21, and the water pipe is connected to the water tank through a pump, and a valve is installed on one end of the pump that is away from the water tank. The water injection assembly only completes the work of injecting the fluid medium into the pressurized chamber 2, and does not perform cleaning work.

[0054] 7 and 8, the drive assembly 4 in this embodiment includes a threaded shaft 41, a drive ring seat 42, a compression block 43, and a feed screw block 44. The threaded shaft 41 is fixedly connected to one end of the moving chamber 22 extending from the fixed chamber 21, and the threaded shaft 41 and the moving chamber 22 share a central axis. A first shaft seat 45 is fixedly connected to the base 1, and the drive ring seat 42 is rotatably connected to the first shaft seat 45 and shares a central axis with the moving chamber 22. A drive part 46 is provided on the base 1. The drive part 46 in this embodiment includes a first electric motor 461 fixedly attached to the base 1 and a drive gear 462 fixedly connected to the drive end of the first electric motor 461. The drive gear 462 meshes with the ring outer wall of the drive ring seat 42. One end of the threaded shaft 41 away from the moving chamber 22 penetrates the inner ring wall of the driving ring seat 42, and there is a gap between the outer peripheral wall of the threaded shaft 41 and the inner ring wall of the driving ring seat 42. In this embodiment, a guide block 47 is connected to the base 1 so as to be slidable in the axial direction of the threaded shaft 41, and an interlocking assembly 5 is provided between the guide block 47 and the threaded shaft 41.

[0055] As shown in Figures 8 and 9, in this embodiment, there are four feed screw blocks 44, all of which are connected to the inner ring wall of the drive ring seat 42 so as to be slidable in the radial direction of the drive ring seat 42, and the four feed screw blocks 44 are uniformly distributed in the circumferential direction of the drive ring seat 42 and arranged on the same center of a circle; in this embodiment, there are four compression blocks 43, all of which are connected to the inner ring wall of the drive ring seat 42 so as to be slidable in the radial direction of the drive ring seat 42, and the four compression blocks 43 are uniformly distributed in the circumferential direction of the drive ring seat 42 and arranged on the same center of a circle; the four feed screw blocks 44 are located on one side of the four compression blocks 43 away from the moving chamber 22, and the compression blocks 43 and the feed screw blocks 44 have opposite sliding directions; and the drive ring seat 42 is provided with a power assembly 6 for driving the sliding movement of the feed screw blocks 44 and the compression blocks 43.

[0056] The interlocking assembly 5 is started, the guide block 47 is fixed relative to the threaded shaft 41, the power assembly 6 is started, the four feed screw blocks 44 are brought into contact with the threaded shaft 41 and engaged with the threads of the threaded shaft 41, and at the same time, the compression block 43 is separated from the threaded shaft 41. The driving part 46 is started, the driving ring seat 42 is rotated, and the threaded shaft 41 generates a yarn feed between the feed screw block 44 and the threaded shaft 41, and the threaded shaft 41 can be driven to make the moving chamber 22 slide in the axial direction. The interlocking assembly 5 is operated, the guide block 47 is separated from the threaded shaft 41, and the power assembly 6 is started, so that the feed screw block 44 is separated from the threaded shaft 41, and at the same time, the compression block 43 is brought into close contact with the threaded shaft 41, at this time the driving ring seat is rotated, and the compression block 43 can drive the threaded shaft 41 to rotate in place, and thus the moving chamber 22 to rotate in place.

[0057] 8 and 9, the power assembly 6 includes a driving ring gear 61, a first gear 62, a second gear 63, a first screw block 64, a second screw block 65, a first screw insert 66, and a second screw insert 67. A receiving cavity 68 is provided in the driving ring seat 42. The driving ring gear 61 is rotatably connected to the cavity wall of the receiving cavity 68 and shares a central axis with the driving ring seat 42. The driving ring gear 61 is located on the circumferential side of the threaded shaft 41 and between the compression block 43 and the feed screw block 44. A rotary drive part 69 is provided on the cavity wall of the receiving cavity 68. The rotary drive part 69 includes a second electric machine 691 fixedly connected to the cavity wall of the receiving cavity 68 and a rotary drive gear 692 fixedly connected to the second electric machine 691. The rotary drive gear 692 meshes with the ring inner wall of the driving ring gear 61. The first gears 62 and the second gears 63 are both rotatably connected to the cavity wall of the accommodation cavity 68, the first gears 62 and the feed screw block 44 are provided in one-to-one correspondence, and the second gears 63 and the compression block 43 are provided in one-to-one correspondence. The four first gears 62 and the four second gears 63 mesh with the outer ring wall of the drive ring gear 61.

[0058] The first thread insert 66 is connected to the first gear 62 via a first bevel gear set 7, which includes a first drive bevel gear 72 coaxially connected to the first gear 62 and a first reverse bevel gear 71 fixedly connected to the first thread insert 66, and the first reverse bevel gear 71 meshes with the first drive bevel gear 72. The first thread block 64 is fixedly connected to one end extending into the receiving cavity 68 of the feed screw block 44, and the first thread block 64 is threadedly connected to the inner peripheral wall of the first thread insert 66. The second threaded insert 67 is connected to the second gear 63 via a second bevel gear set 8, which includes a second driving bevel gear 82 coaxially connected to the second gear 63 and a second reversing bevel gear 81 fixedly connected to the second threaded insert 67, and the second reversing bevel gear 81 meshes with the second driving bevel gear 82. The second threaded block 65 is fixedly connected to one end extending into the receiving cavity 68 of the compression block 43, and the second threaded block 65 is threadedly connected to the inner peripheral wall of the second threaded insert 67.

[0059] The second electric machine 691 is driven to rotate the driving ring gear 61 in the forward direction, so that the first gear 62 and the second gear 63 rotate, the first gear 62 drives the rotation of the first screw insert 66 through the first bevel gear set 7, the second gear 63 drives the rotation of the second screw insert 67 through the second bevel gear set 8, the first screw insert 66 and the first screw block 64 generate yarn feed, the second screw insert 67 and the second screw block 65 generate yarn feed, and the first screw block 64 and the second screw block 65 have opposite yarn feed directions, so that the compression block 43 and the feed screw block 44 slide in opposite directions, the compression block 43 moves away from the threaded shaft 41, and the feed screw block 44 abuts against the threaded shaft 41. Similarly, when the driving ring gear 61 is reversed, the compression block 43 is brought into close contact with the threaded shaft 41, and the feed screw block 44 moves away from the threaded shaft 41.

[0060] As shown in Figures 10 and 11, the interlocking assembly 5 includes a support block 51, a press block 52, a lifting screw 53 and a lifting screw ring 54. The support block 51 is fixedly connected to the top end of the guide block 47. A first arc trough 55 is opened on the upper surface of the support block 51. The groove wall of the first arc trough 55 is smooth and the groove wall of the first arc trough 55 is in tension with the threaded shaft 41. The press block 52 is vertically slidably mounted on the support block 51 and is located above the threaded shaft 41. The press block 52 has a second arc trough 55 on the lower surface. A trough 56 is opened, and the groove wall of the second arc trough 56 is rough. The lifting screw 53 is fixedly connected to the bottom end of the press block 52. A recess 57 is provided on the upper surface of the support block 51. The bottom end of the lifting screw 53 extends into the recess 57. A third electric motor 58 is fixedly connected to the support block 51. A power gear 59 is fixedly connected to the driving end of the third electric motor 58. The lifting screw ring 54 is rotatably connected to the support block 51 and is screwed to the lifting screw 53. The power gear 59 meshes with the outer ring wall of the lifting screw ring 54. By driving the third electric motor 58, the power gear 59 meshes with the lifting screw ring 54, the lifting screw 53 and the lifting screw ring 54 generate yarn feed, the press block 52 moves downward, and the second arc trough 56 comes into close contact with the threaded shaft 41, so that the guide block 47 can be fixed relatively to the threaded shaft 41.

[0061] The working principle of the present embodiment 3 is that, at the beginning, the moving chamber 22 is separated from the fixed chamber 21, the carrier cover 26 is mostly extended from the guide sleeve 25 and is in a fixed position, and the packaging bag is put into the carrier cover 26 according to the decellularization parameters in embodiment 1. The second electric motor 691 is driven, the compression block 43 is separated from the threaded shaft 41, the feed screw block 44 is abutted against the threaded shaft 41, the third electric motor 58 is started, the press block 52 moves downward, the second arc trough 56 is in close contact with the threaded shaft 41, the driving part 46 is driven, the driving ring seat 42 rotates, the threaded shaft 41 drives the moving chamber 22 to move axially to the fixed chamber 21, and the moving chamber 22 is inserted into the fixed chamber 21, and in this process, the guide ball 29 rolls in the linear arc groove 262.

[0062] After the moving chamber 22 reaches the predetermined position, the second electric motor 691 is driven, the compression block 43 is tightly attached to the threaded shaft 41, the feed screw block 44 is separated from the threaded shaft 41, the driving element 46 is started, the driving ring seat 42 is rotated, and the threaded shaft 41 is driven to rotate the moving chamber 22 on the spot, at this time, the guide ball 29 enters the spiral arc groove 261, the spherical wall of the guide ball 29 presses against the groove wall of the spiral arc groove 261 and rolls in the spiral arc groove 261, the carrier cover 26 moves axially to the guide sleeve 25, and after the carrier cover 26 reaches the predetermined position, the guide ball 29 moves into the linear arc groove 262, at this time, the driving element 46 is further started, the moving chamber 22 is positioned at the appropriate position of the fixed chamber 21, and the loading of the product is completed.

[0063] When the water pump 391 is started, the fluid medium is introduced into the fixed chamber 21, and after there are no bubbles in the pressurized chamber 2, the second control valve 212 and the first control valve 392 are closed, the water pump 391 is closed, the injection of the fluid medium is completed, and then the high pressure treatment is performed. After the treatment is completed, the fluid medium is discharged, the moving chamber 22 is separated from the fixed chamber 21, and during the moving process of the moving chamber 22, the carrier cover 26 extends from the guide sleeve 25, and then the packaging bag is removed to complete the high pressure treatment.

[0064] Performance Detection The performance of the decellularized tissues obtained in Examples 1 and 2 was examined, and the results are specifically as follows.

[0065] 1. Calculation of decellularized DNA ratio: The mass of each decellularized tissue is measured and prepared as a test piece for preparation.

[0066] 1. Measurement of DNA content of test piece: The test piece is immersed in a protein decomposition enzyme solution to dissolve it, then treated with phenol / chloroform to remove proteins, and DNA is recovered by ethanol precipitation. The recovered DNA is fluorescently stained with PicoGreen (Life Technologies), and the DNA is quantified by measuring the fluorescence intensity. The DNA content of the test piece is calculated from the mass of the test piece and the amount of DNA.

[0067] (DNA content of test strip) = (amount of DNA) / (mass of test strip).

[0068] (2) Measurement of DNA content in dried test specimens: After holding the test specimens in a constant temperature bath at 60°C for 12 hours, their masses are detected and the rate of loss on drying is calculated. The DNA content per unit of dry mass of the test specimens is calculated from the DNA content and rate of loss on drying of the test specimens.

[0069] (Loss on drying) = (mass of dried test piece) / (mass of test piece).

[0070] (DNA content of dried test specimen) = (DNA content of test specimen) / (loss on drying).

[0071] 2. Calculate the decellularized DNA ratio.

[0072] According to the solutions in step (1) and step (2), detect the DNA content of the dry specimen of non-decellularized tissue. The calculation method of the decellularized DNA ratio is as follows:

[0073] (Decellularized DNA ratio)=(DNA content of dried specimen of decellularized tissue) / (DNA content of dried specimen of non-decellularized tissue).

[0074] 2. Cell migration test Starvation was performed by culturing L929 cells (mouse fibroblasts) in MEM medium at 37°C for 24 hours. 1 mL of MEM medium containing 5% decellularized tissue was added to a 24-well culture dish, and a cell culture insert (pore size 8 μm) was placed in each well. The L929 cells cultured in MEM medium were then transferred to the insert (1.0x10 5 The cells were seeded at 1000 x 1000 cells / well and cultured at 37° C. for 3 hours, and the number of cells in the wells that migrated to the bottom of the insert was measured. The cell number was measured using a fluorescent microscope after DAPI staining, and the average number N of five wells was calculated.

[0075] 1. Neurite Outgrowth Test PC12 cells (pheochromocytoma derived from rat adrenal medulla) were cultured in collagen-coated 24-well culture dishes (1.0x10 4 The cells are seeded at 1000 x 1000 cells / well) and cultured at 37°C for 24 hours. The medium is switched to RPMI medium containing 0.1% horse serum, and a cell culture insert (pore size 8 μm) is placed in each well. 200 μL of saline containing 5% decellularized tissue is added to the insert, and the culture is continued for 24 hours with and without the addition of 50 ng of nerve growth factor. The cells are observed using a phase contrast microscope, and the effect of neurite outgrowth is estimated according to the following criteria:

[0076] 2. Decellularization Stability Detection According to the decellularization steps of Example 1 and Example 2, 20 decellularization processes are performed, and then the decellularized DNA ratio is calculated, and a cell migration test and a neurite outgrowth test are performed on the 20 sets of decellularized tissues of Example 1 and the 20 sets of decellularized tissues of Example 2. Using the first set of decellularized tissue as a standard, the performance deviation values ​​of the other sets of decellularized tissues are detected, and the performance deviation value = (|(n performance of Example)-(performance of Example 1)|) / (performance of Example 1) is detected, and finally the average value is recorded.

[0077] The detection results are shown in Table 1. Decellularized tissue performance detection table JPEG2025515929000002.jpg85146

[0078] First, as can be seen from the detection results, whether it is blood vessels or pig bone marrow, the decellularized tissue obtained in this application has low cytotoxicity and excellent cell induction differentiation effect. Furthermore, whether it is in Example 1 or Example 2, after performing the same decellularization steps 20 times, the performance deviation of the final decellularized tissue does not exceed 0.01%, indicating that the decellularization process of this application has good stability.

[0079] The specific examples are merely illustrative of the present application and do not limit the present application. After reading this specification, a person skilled in the art may make amendments to the present examples as necessary without making any creative contribution thereto, but as long as they are within the scope of the claims of this application, they shall be protected by the Patent Law. [Explanation of symbols]

[0080] 1 Base 11 First Base Block 2. Pressurized chamber 21 Fixed Chamber 211 Exhaust pipe 212 Second control valve 213 Drainage pipe 214 Third control valve 22 Mobile Chamber 221 Second Base Block 23 First Seal 24 First link 25 Guide sleeve 26 Carrier cover 261 Spiral Arc Groove 262 Straight arc groove 27 Magnetic layer 28 Water channel 29 Guide Ball 291 Ring Set 3 Water Injection Assembly 31 Water tank 32 Water inlet pipe head 321 Water Inlet 322 Second Seal 33 Water outlet pipe head 34 Communication pipe 35 Spray ball head 351 Water passage 36 Mounting groove 361 No. 1 Pipe Trough 362 Second Pipe Trough 37 3rd Base Block 38 Sliding parts 39 Water pipe 391 Water Pump 392 First control valve 4 Drive Assembly 41 Threaded Shaft 42 Drive ring seat 43 Compressed Blocks 44 Lead screw block 45 1st shaft seat 46 Drive parts 461 First Electric 462 Drive Gear 47 Guide Block 5 Interlocking Assembly 51 Support Block 52 Press Block 53 Lifting screw 54 Lifting screw ring 55 First Arc Trough 56 Second Arc Trough 57 Undercut 58 Third Electric 59 Power Gear 6 Power Assembly 61 Drive ring gear 62 1st Gear 63 2nd Gear 64 First Screw Block 65 Second screw block 66 First thread insert 67 2nd thread insert 68 Containment Cavity 69 Rotary drive parts 691 Second Electric 692 Rotary Drive Gear 7 1st bevel gear set 71 1st reversing bevel gear 72 1st drive bevel gear 8 No. 2 bevel gear set 81 2nd reversing bevel gear 82 Second driving bevel gear

Claims

1. 1. A method for preparing biological tissue for transplantation, comprising the steps of: Removing tissue; A pre-cleaning step; a high-pressure treatment in which the tissue and a treatment solution are placed together in a packaging bag, the packaging bag is sealed, the packaging bag is then placed in a fluid medium, and a water purification pressure is applied to the packaging bag, the high-pressure treatment comprising components of the treatment solution including a nucleic acid degrading enzyme and a buffer solution; and post-cleaning the tissue after high pressure treatment. A method for processing biological tissue for transplantation, comprising:

2. The buffer solution includes at least one of physiological saline, a PBS buffer solution, and a HEPES buffer solution.

2. The method for preparing biological tissue for transplantation according to claim 1.

3. In the high pressure treatment, the packaging bag is at least two-layered, the tissue and the treatment liquid are located in the innermost packaging bag, and the remaining packaging bags are filled with a liquid medium.

2. The method for preparing biological tissue for transplantation according to claim 1.

4. In the high pressure treatment, the packaging bag is double-layered, the tissue and the treatment liquid are located in the innermost packaging bag, and the outer packaging bag is filled with a liquid medium, and the density of the treatment liquid is greater than that of the liquid medium.

4. The method for preparing biological tissue for transplantation according to claim 3.

5. In the high-pressure treatment step, the packaging bag is placed in a high-voltage device to perform high-pressure treatment. The high-voltage device includes a base (1), a pressurized chamber (2) and a water injection assembly (3). The pressurized chamber (2) includes a fixed chamber (21) and a moving chamber (22). The fixed chamber (21) is fixedly connected to the base (1). The inner wall of the fixed chamber (21) is a peripheral wall that opens at one axial end. The outer periphery of the moving chamber (22) is a peripheral wall that opens at one axial end. The opening of the moving chamber (22) is a The end of the movable chamber (22) is inserted into the fixed chamber (21), the outer peripheral wall of the movable chamber (22) and the inner peripheral wall of the fixed chamber (21) are stuck together, a first seal (23) is provided at the open end of the fixed chamber (21), and the first seal (23) and the outer peripheral wall of the movable chamber (22) are abutted against each other, the base (1) is provided with a drive assembly (4) for driving the movable chamber (22) to slide or rotate in place, a guide sleeve (25) is fixedly connected to the inner wall of the fixed chamber (21), and the drive assembly (4) is provided with a guide sleeve (25) fixedly connected to the inner wall of the fixed chamber (21). A carrier cover (26) is slidably mounted within the inner sleeve (25). A water passage groove (28) is formed through both the guide sleeve (25) and the carrier cover (26). A spiral arc groove (261) is formed on the outer peripheral wall of the carrier cover (26). A linear arc groove (262) is further formed on the outer peripheral wall of the carrier cover (26). The linear arc groove (262) penetrates the carrier cover (26) in the axial direction of the carrier cover (26). The linear arc groove (262) and the spiral arc groove (263) are aligned in a line. The carrier cover (26) has one end extending from the guide sleeve (25) located within the moving chamber (22), a guide ball (29) is rotatably connected to the inner wall of the moving chamber (22), the guide ball (29) and the linear arc groove (262) are provided in one-to-one correspondence, the guide ball (29) has one end remote from the moving chamber (22) inserted into the linear arc groove (262) or the spiral arc groove (261), and the spherical wall of the guide ball (29) isabutting against the groove wall of the linear arc groove (262) or the groove wall of the spiral arc groove (261), and the water injection assembly (3) and the fixed chamber (21) are provided through each other; 2. The method for preparing biological tissue for transplantation according to claim 1.

6. The inner peripheral wall of the guide sleeve (25) and the outer peripheral wall of the carrier cover (26) are both provided with a magnetic layer (27).

6. The method for preparing biological tissue for transplantation according to claim 5.

7. The drive assembly (4) comprises a threaded shaft (41), a drive ring seat (42), a compression block (43) and a feed screw block (44), the threaded shaft (41) is fixedly connected to one end of the moving chamber (22) extending from the fixed chamber (21), the drive ring seat (42) is rotatably connected to the base (1), the base (1) is provided with a drive part (46) for driving the rotation of the drive ring seat (42), and the threaded shaft (41) is fixed to one end of the moving chamber (22) extending from the fixed chamber (21). One end of the threaded shaft (41) that is away from the moving chamber (22) passes through the inner ring wall of the driving ring seat (42), and there is a gap between the outer peripheral wall of the threaded shaft (41) and the inner ring wall of the driving ring seat (42). At least three of the feed screw blocks (44) are connected to the inner ring wall of the driving ring seat (42) so as to be slidable in the radial direction of the driving ring seat (42), and the at least three feed screw blocks (44) are uniformly distributed in the circumferential direction of the driving ring seat (42), and the feed screw blocks (44) is abutted against the threaded shaft (41) at one end of the threaded shaft (41) and screw-connected thereto, and at least three of the compression blocks (43) are connected to the ring inner wall of the drive ring seat (42) so as to be slidable in the radial direction of the drive ring seat (42), and there is a gap between the compression block (43) and the threaded shaft (41) at one end of the threaded shaft (41), and the at least three compression blocks (43) are uniformly distributed in the circumferential direction of the drive ring seat (42). The at least three compression blocks (43) are arranged in a direction opposite to the sliding movement direction of the at least three feed screw blocks (44), the drive ring seat (42) is provided with a power assembly (6) for driving the sliding movement of the feed screw blocks (44) and the compression blocks (43), the base (1) is provided with a guide block (47) slidably movable in the axial direction of the threaded shaft (41), and an interlocking assembly (5) is provided between the guide block (47) and the threaded shaft (41).

6. The method for treating biological tissue for transplantation according to claim 5.

8. The power assembly (6) includes a driving ring gear (61), a first gear (62), a second gear (63), a first screw block (64), a second screw block (65), a first screw insert (66) and a second screw insert (67). A receiving cavity (68) is provided in the driving ring seat (42). The driving ring gear (61) is rotatably connected to a cavity wall of the receiving cavity (68). A rotary drive part (69) for driving the rotation of the driving gear (462) is provided on the cavity wall of the receiving cavity (68). The first gear (62) and the second gear (63) are both rotatably connected to the cavity wall of the receiving cavity (68) and are both meshed with the driving ring gear (61). The first gear (62) and the feed screw block (44) correspond to each other one-to-one. the second gear (63) and the compression block (43) are provided in one-to-one correspondence, the first thread insert (66) is connected to the first gear (62) via a first bevel gear set (7), the first thread block (64) is fixedly connected to one end of the feed screw block (44) extending into the receiving cavity (68), and the first thread block (64) and the inner peripheral wall of the first thread insert (66) are threadedly connected to each other, the second thread insert (67) is connected to the second gear (63) via a second bevel gear set (8), the second thread block (65) is fixedly connected to one end of the compression block (43) extending into the receiving cavity (68), and the second thread block (65) and the inner peripheral wall of the second thread insert (67) are threadedly connected to each other.

8. The method for preparing biological tissue for transplantation according to claim 7.

9. The water injection assembly (3) comprises a water supply tank (31), a water inlet pipe head (32), a water outlet pipe head (33), a connecting pipe (34) and a spray ball head (35). The fixed chamber (21) has an attachment groove (36) on its outer wall away from the moving chamber (22). The fixed chamber (21) has a first pipe trough (361) and a second pipe trough (362) on its inner wall opposite to the open end. The first pipe trough (361) and the second pipe trough (362) are both passed through the attachment groove (36). The water inlet The pipe head (32) is slidably mounted on the groove wall of the mounting groove (36), and a sliding part (38) for driving the sliding movement of the water inlet pipe head (32) is provided on the base (1). The water inlet pipe head (32) is mounted with one end located within the mounting groove (36) sealed. A water supply pipe (39) and a water pump (391) are provided between the one end of the water inlet pipe head (32) extending from the mounting groove (36) and the water supply tank (31). The water supply pipe (39) is fixed to the water pump (391). A first control valve (392) is provided at one end close to the chamber (21), one end of the first pipe trough (361) which passes through the mounting groove (36) and one end of the second pipe trough (362) which passes through the mounting groove (36) are arranged side by side in the sliding movement direction of the water inlet pipe head (32), a water passage port (321) is provided through the peripheral wall of the water inlet pipe head (32), the water outlet pipe head (33) is attached to the groove wall of the first pipe trough (361), and the communicating pipe (34) is provided in the groove wall of the second pipe trough (362). A second seal (322) is provided at one end of the communicating pipe (34) close to the water inlet pipe head (32) and at one end of the water outlet pipe head (33) close to the water inlet pipe head (32). The groove wall of the water port (321) passes through the water outlet pipe head (33) or passes through the communicating pipe (34). The spray ball head (35) is fixedly connected to the inner wall facing the open end of the fixed chamber (21). A number of water passages (351) are provided in the spray ball head (35). The water passages (351) areThe spray ball head (35) penetrates the spherical wall extending to the fixed chamber (21), and one end of the communication pipe (34) remote from the water inlet pipe head (32) is connected to the spray ball head (35) and passes through all of the water passages (351). An exhaust pipe (211) is connected to the top end of the fixed chamber (21), and a second control valve (212) is attached to the exhaust pipe (211).

6. The method for preparing biological tissue for transplantation according to claim 5.

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