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] This application relates to the field of tissue transplantation, and more particularly to methods for preparing biological tissue for transplantation. [Background technology]

[0002] Tissue transplantation 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 typically include bone and cartilage transplants, skin and fat transplants, mucosal transplants, fascia transplants, muscle transplants, nerve transplants, composite tissue transplants, biomaterial implants, 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 materials and bio-derived materials is gradually becoming a more common option for tissue transplantation, but the body usually tends to reject artificial materials. To solve the above problem, decellularization technology, which is a method for processing bio-derived materials, has recently attracted attention.

[0004] Decellularization techniques use chemical and physical methods to remove cells from allogeneic or xenogeneic tissues, resulting in the creation of biological tissue scaffolds with low or no immunogenicity. Methods for decellularizing biological tissues include the use of surfactants, enzymes, acidifiers, and ultra-high pressure hydrostatic pressure. Ultra-high pressure hydrostatic pressure treatment, which does not use toxic chemicals, produces decellularized tissue that is less cytotoxic than chemical methods, meaning that the remaining active substances have a superior effect on inducing host cell differentiation after transplantation compared to chemical methods.

[0005] However, the inventors have found that, although temperature and pressure must be controlled when processing cell tissues using ultra-high hydrostatic pressure, the properties of the decellularized tissue ultimately obtained under the same temperature and pressure processing conditions may not be consistent. This indicates that ultra-high hydrostatic pressure can 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 tissue treatment, resulting in decellularized tissue with low cytotoxicity and excellent cell differentiation induction effects. This 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 uses the following technical solutions. 1. 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.The high-pressure treatment includes high-pressure treatment in which the components of the treatment solution include a nucleic acid degrading enzyme and a buffer solution, and post-washing in which the tissue after the high-pressure treatment is washed.

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

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

[0010] The inventor conducted numerous experiments and studies, but was unable to find a way to improve the stability of the decellularization process. Ultimately, the inventor concluded that the process of obtaining decellularized tissue using ultra-high hydrostatic pressure must be carried out in a fluid medium. While intuitively considering changing the type of fluid medium, he devised a different approach to solve the problem. The inventor pre-packaged the biological tissue in a packaging bag, and placed a mixture of nuclease and buffer solution in the packaging bag as the treatment solution, isolating the biological tissue and treatment solution from the harsh environment of the ultra-high hydrostatic pressure treatment. Ultimately, the inventor found that this procedure significantly improved the stability of the ultra-high hydrostatic pressure in the decellularization process. Furthermore, by placing the treatment solution containing the nuclease and buffer solution in the packaging bag together with the biological tissue, the efficiency of the biological tissue treatment was also significantly improved. The resulting decellularized tissue exhibited low cytotoxicity and excellent cell induction differentiation effects.

[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 obtained can be 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 solution, the inventors discovered that the decellularization process is more stable when the density of the processing liquid is greater than the density of the liquid medium, and the inventors speculated that this is because, in the process of pressure being transmitted to biological cell tissue, the density of the processing liquid is greater, which can reduce the impact of the large environmental pressure of ultra-high purified water on the small environmental pressure of the packaging bag, allowing pressure to be applied to the biological tissue more stably and uniformly.

[0016] Preferably, in the high-pressure treatment step, the packaging bag is placed in a high-voltage device for high-pressure treatment, the high-voltage device comprising a base, a pressurizing chamber and a water injection assembly, the pressurizing chamber including a fixed chamber and a movable chamber, the fixed chamber being fixedly connected to the base, the inner wall of the fixed chamber being a peripheral wall and opening at one end in the axial direction, the outer periphery of the movable chamber being a peripheral wall and opening at one end in the axial direction, the open end of the movable chamber being inserted into the fixed chamber, the outer periphery wall of the movable chamber and the inner periphery wall of the fixed chamber being attached to each other, a first seal being provided at the open end of the fixed chamber and the first seal being abutted against the outer periphery wall of the movable chamber, the base being provided with a drive assembly for driving the movable chamber to slide or rotate on the spot, a guide sleeve being fixedly connected to the inner wall of the fixed chamber, and a carrier cover being provided within the guide sleeve. the guide sleeve and the carrier cover are slidably mounted, and both have water-passing grooves extending therethrough; the carrier cover has an outer peripheral wall formed with a spiral arc groove and a linear arc groove further formed therein, the linear arc groove passing through the carrier cover in the axial direction of the carrier cover, and the linear arc groove and the spiral arc groove are arranged through each other; one end of the carrier cover extending from the guide sleeve is located within the movable chamber, and a guide ball is rotatably connected to the inner wall of the movable chamber, and the guide ball and the linear arc groove are arranged in one-to-one correspondence, and one end of the guide ball remote from the movable chamber is inserted into the linear arc groove or the spiral arc groove, and the spherical wall of the guide ball abuts against the groove wall of the linear arc groove or the groove wall of the spiral arc groove; the water-injection assembly and the fixed chamber are arranged 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 products can be easily stored and removed. The driving assembly allows the moving chamber to slide axially, and the guide ball rolls in the linear arc groove, allowing the moving chamber to slide axially with the fixed chamber, facilitating the tightening of the pressurizing chamber. When the driving assembly drives the carrier cover to rotate in place, the guide ball enters the spiral arc groove accordingly, creating a movement similar to thread feeding between the carrier cover and the moving chamber. The carrier cover can slide on the guide sleeve, and the carrier cover can move in and out of the guide sleeve and then in and out of the fixed chamber. Therefore, the guide ball, linear arc groove, and spiral arc groove of the present application cooperate with the carrier cover and the moving chamber to achieve axial sliding movement of the moving chamber and simultaneously achieve sliding movement of the carrier cover, making it easy to store the moving chamber and carrier cover, and for the operator to easily remove 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 comprises a threaded shaft, a drive ring seat, a compression block, and a lead screw block, the threaded shaft being fixedly connected to one end of the moving chamber extending from the fixed chamber, the drive ring seat being rotatably connected to the base, and the base being 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 being passed through the inner ring wall of the drive ring seat, and 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 of the lead screw blocks being 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, the at least three lead screw blocks being uniformly distributed in the circumferential direction of the drive ring seat, and the lead screw blocks a feed screw block abutting against one end of the threaded shaft and threadedly connected to the threaded shaft; at least two compression blocks 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 gap is formed between the compression blocks and the threaded shaft at one end of the threaded shaft; at least three compression blocks are uniformly distributed in the circumferential direction of the drive ring seat; and 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 the 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 drive ring seat rotates, the meshing effect drives the first gear and the second gear to rotate, and then the direction-changing action of the first bevel gear set and the second bevel gear set rotates the first thread insert and the second thread insert, thereby generating yarn feeding between the first thread insert and the first screw block, and between the second thread insert and the second screw 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 opposite directions, so that the compression block is tightly attached to 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 separates from the threaded shaft, and the feed screw block abuts against the threaded shaft and is thus engaged with the threads on the threaded shaft. At the same time, the guide block and the threaded shaft are fixed relative to each other by the interlocking assembly, so that when the driving ring seat rotates, a thread feed occurs between the feed screw block and the threaded shaft, and the threaded shaft drives the moving chamber to slide axially, which is convenient and fast.

[0022] Preferably, the power assembly includes 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 screw block is fixedly connected to one end of the feed screw block extending into the accommodating cavity, and the first screw block and the inner wall of the first thread insert are threadedly connected to each other, the second thread insert is connected to the second gear via a second bevel gear set, the second screw block is fixedly connected to one end of the compression block extending into the accommodating cavity, and the second screw block and the inner wall of the second thread insert are threadedly connected to each other.

[0023] Preferably, the water injection assembly comprises 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 attachment groove formed in an outer wall away from the movable chamber; the fixed chamber has a first pipe trough and a second pipe trough formed on an inner wall opposite the open end; the first pipe trough and the second pipe trough are both passed through the attachment groove; the water inlet pipe head is slidably mounted on the groove wall of the attachment 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 attachment 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 attachment 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 passing through the attachment groove and one end of the second pipe trough passing through the attachment groove are are arranged side by side in the sliding direction of the water inlet pipe head, a water passage hole 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 connecting pipe is attached to the groove wall of the second pipe trough, one end of the connecting 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 hole passes through the water outlet pipe head or the connecting pipe, the spray ball head is fixedly connected to the inner wall opposite the open end of the fixed chamber, several water passages are formed within the spray ball head, the water passages pass through the spherical wall of the spray ball head extending to the fixed chamber, one end of the connecting 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 is connected to the water outlet pipe head, the water tank, water pipe, and water pump cooperate with the water inlet and outlet pipe heads to properly inject the fluid medium into the pressurized chamber. After the high-pressure treatment is completed, the water inlet pipe head is moved to connect the water inlet to the connecting pipe, and the fluid medium or cleaning water enters the spray ball head and is guided by the water passage. The fluid medium or cleaning water is sprayed at multiple angles into the fixed chamber, cleaning the fixed chamber and the moving chamber to a certain extent, thereby improving the cleanliness of the decellularized tissue treatment process. [Effects of the Invention]

[0025] In summary, the present application has the following beneficial effects: 1. In this application, a mixture of nuclease and buffer solution is used as the treatment solution, and then the treatment solution and biological tissue are pre-packaged and placed in a fluid medium of a high-voltage device, where the biological tissue and treatment solution are relatively isolated from the large environment of the ultra-high hydrostatic pressure treatment, and the biological tissue is subjected to high-pressure treatment at a temperature of 20-25°C. This significantly improves the stability of the ultra-high hydrostatic pressure in the decellularization process of cellular tissue, and significantly improves the efficiency of biological tissue treatment, and the resulting decellularized tissue has low cytotoxicity and excellent cell-induced differentiation effects.

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

[0027] 3. This application provides a high voltage device that allows convenient and quick storage and removal of products. [Brief explanation 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. [Figure 2] FIG. 2 is a cross-sectional view taken along line AA in FIG. [Figure 3]FIG. 3 is an exploded view showing a moving chamber structure in Example 3 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. [Figure 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 Example 3 of the present application. [Figure 10] FIG. 10 is a schematic diagram showing an interlocking assembly structure in Example 3 of the present application. [Figure 11] FIG. 11 is a cross-sectional view taken along line EE in FIG. DETAILED DESCRIPTION OF THE INVENTION

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

[0030] Example Example 1 In this Example 1, a method for processing 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 swine at 4°C into 1 cm long segments.

[0032] Step 2: Pre-wash Physiological saline is used as a washing solution, and the excised pig 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 solution are placed together in a first packaging bag, which is then clad and sealed. The packaging bag containing the vascular tissue is then placed in a second packaging bag, and a liquid medium is added to the second packaging bag. The second packaging bag is then clad and sealed. In this embodiment, there are no bubbles between the two packaging bags.

[0034] Finally, the packaged two-layer packaging bag is placed in a high-pressure processing 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, and the pressure application time is set to 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, and the liquid medium is water.

[0036] Step 4: After washing, The vascular tissue after high-pressure treatment was 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 then washed with HEPES buffer solution with oscillation for 2 weeks to obtain decellularized tissue. The cleaning solution was the same substance as the treatment solution in this example.

[0037] Example 2 In this Example 1, a method for processing 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 edible pigs is stored at 4°C in small blocks measuring 1cm x 1cm.

[0039] Step 2: Pre-wash Physiological saline is used as a washing solution, and the excised pig 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 treatment liquid are placed together in a first packaging bag, which is then clad and sealed. The packaging bag containing the pig bone marrow tissue is then placed in a second packaging bag, and a liquid medium is added to the second packaging bag. The second packaging bag is then clad and sealed. In this embodiment, there are no bubbles between the two packaging bags.

[0041] Finally, the packaged two-layer packaging bag is placed in a high-pressure processing 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, and the pressure application time is set to 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 the nuclease to the HEPES buffer solution is 1:9.3, and the liquid medium is water.

[0043] Step 4: After washing, The high-pressure treated pig bone marrow tissue is placed in a washing solution and washed by oscillation for 2 weeks at 37°C, after which decellularized tissue is obtained. The washing solution is the same substance as the washing solution in this example.

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

[0045] 1 and 2, the high-voltage equipment comprises 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 movable chamber 22. One axial end of the fixed chamber 21 and one axial end of the movable 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 has a horizontal axial direction. The open end of the movable chamber 22 is inserted into the fixed chamber 21, and the outer peripheral wall of the movable chamber 22 and the inner peripheral wall of the fixed chamber 21 are attached to each other. The movable chamber 22 may be able to slide axially within the fixed chamber 21, or may be able to rotate circumferentially. 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 abuts against the outer wall of the movable chamber 22. A drive assembly 4 is provided on the base 1 to drive the rotation or sliding movement of the movable 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 the inner peripheral wall of the fixed chamber 21 on one side 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 formed on the outer peripheral wall of the carrier cover 26, and two straight arc grooves 262 are further formed 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 uniformly distributed in the circumferential direction of the carrier cover 26. 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 located inside the movable chamber 22, a ring set 291 is fixedly connected to the inner wall of the movable chamber 22 via a second base block 221, the ring set 291 and the linear arc groove 262 are arranged in one-to-one correspondence, a guide ball 29 is rotatably connected inside 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 the other end of the guide ball 29 extending from the ring set 291 is inserted into the linear arc groove 262, and the spherical wall of the guide ball 29 is in contact with the groove wall of the linear arc groove 262.

[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. The product is placed on the carrier cover 26, and then the drive assembly 4 is activated, and the moving chamber 22 moves axially toward the fixed chamber 21. The moving chamber 22 is inserted into the fixed chamber 21, and the guide ball 29 rolls in the linear arc groove 262. After the movable chamber 22 reaches the predetermined position, the driving assembly 4 drives the movable chamber 22 to rotate on the spot. At this time, the guide ball 29 enters the spiral arc groove 261, and the spherical wall of the guide ball 29 presses against the groove wall of the spiral arc groove 261 while rolling along the spiral arc groove 261, causing the carrier cover 26 to move axially toward the guide sleeve 25. After the carrier cover 26 reaches the predetermined position, the guide ball 29 moves to the linear arc groove 262. At this time, the driving assembly 4 drives the movable chamber 22 to move axially, allowing the movable chamber 22 to reach the appropriate position in the fixed chamber 21 and completing 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 no bubbles are present in the fixed chamber 21 and the movable chamber 22. Then, the pressurizing chamber 2 is pressurized according to the need. The pressurizing method can be to maintain the volume of the pressurizing chamber 2 as it is and continue to add fluid medium using the water injection assembly 3 to pressurize, or to start the driving assembly 4 to cause the guide ball 29 to continue sliding along the linear arc groove 262, and the movable chamber 22 to slide toward the fixed chamber 21, thereby reducing the volume of the pressurizing 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 drive assembly 4 is started, and the movable chamber 22 reaches a predetermined position in the axial direction away from the fixed chamber 21. Then, the drive assembly 4 drives the movable chamber 22 to rotate on the spot, causing the guide ball 29 to roll in the spiral arc groove 261, thereby extending the carrier cover 26 from the guide sleeve 25. Then, the drive assembly 4 drives the movable chamber 22 to move in the axial direction, causing the guide ball 29 to roll in the linear arc groove 262, thereby separating the movable chamber 22 from the fixed chamber 21. At this time, the product can be removed from the carrier cover 26 exposed to the outside.

[0051] 4 and 5, 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. A rectangular mounting groove 36 is formed on the outer wall of the fixed chamber 21 facing away from the movable chamber 22. The outer peripheral wall of the water inlet pipe head 32 is a rectangular tube that is mounted on the groove wall of the mounting groove 36 so as to be able to slide axially along 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 faces the groove wall of the mounting groove 36. A third base block 37 is fixedly connected to the base 1, and a sliding element 38 is mounted on the third base block 37. The sliding element 38 in this embodiment is a cylinder, and the driving end of the sliding element 38 is fixedly connected to the outer peripheral wall of the water inlet pipe head 32 at one end 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 at the end of the water supply pipe 39 where the water pump 391 is closest to the fixed chamber 21.

[0052] 5 and 6, a first pipe trough 361 and a second pipe trough 362 are arranged side by side on the upper wall of the mounting groove 36 in the axial direction of the fixed chamber 21, with the first pipe trough 361 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 space between the water outlet pipe head 33 and the groove wall of the first pipe trough 361 is sealed and connected, and passes 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, and the water passages 351 penetrate the spherical wall of the spray ball head 35 extending to the fixed chamber 21, 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 of the water passages 351. In this embodiment, a second seal 322 is fixedly connected to one end of the connecting pipe 34 close to the water inlet pipe head 32 and to one end of the water outlet pipe head 33 close to the water inlet pipe head 32. The second seals 322 are sealing rings, and the two second seals 322 are always in contact with the upper wall of the water inlet pipe head 32, through which a water port 321 also penetrates. 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] Initially, the water port 321 passes through the water outlet pipe head 33, and the second seal 322 at the water outlet pipe head 33 is located on the periphery of the water port 321, opening the first control valve 392 and the second control valve 212. At the same time, the second seal 322 at the connecting pipe 34 seals against the water inlet pipe head 32, sealing the connecting pipe 34. The water pump 391 is started, introducing a fluid medium into the fixed chamber 21. After bubbles disappear from the pressurized chamber 2, the second control valve 212 and the first control valve 392 are closed, and the water pump 391 is closed, completing the injection of the fluid medium and allowing high-pressure treatment to begin. After the high-pressure treatment is completed, the third control valve 214 is opened, discharging the fluid medium, and the movable chamber 22 slides out of the fixed chamber 21 to remove the product. After the high-pressure treatment is completed and the product is removed, the sliding element 38 is activated, sliding the water inlet pipe head 32 toward the movable chamber 22, passing the water port 321 through the connecting pipe 34. The second seal 322 of the connecting pipe 34 seals around the water port 321. Cleaning water is then introduced into the water supply tank 31. The first control valve 392 is then opened, and the water pump 391 is activated. The cleaning water enters the water passage 351 through the connecting pipe 34 and is then sprayed into the pressurized chamber 2 via the water passage 351, thereby cleaning the pressurized chamber 2 to a certain extent. 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. The water pipe is connected to the water tank via a pump, and a valve is attached to the end of the pump remote from the water tank. The water injection assembly only completes the task of injecting fluid medium into the pressurized chamber 2; it does not perform cleaning.

[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 movable chamber 22 extending from the fixed chamber 21, and the threaded shaft 41 and the movable 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 movable 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 outer ring wall of the drive ring seat 42. One end of the threaded shaft 41 remote from the moving chamber 22 penetrates the inner ring wall of the drive 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 drive 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 around 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 around 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 slide in opposite directions. A power assembly 6 is provided on the drive ring seat 42 to drive the sliding movement of the feed screw blocks 44 and the compression blocks 43.

[0056] The interlocking assembly 5 is activated to fix the guide block 47 relative to the threaded shaft 41, and the power assembly 6 is activated to bring the four feed screw blocks 44 into contact with the threaded shaft 41 and engage with the threads on the threaded shaft 41. At the same time, the compression block 43 is released from the threaded shaft 41. The drive component 46 is activated to rotate the drive ring seat 42, causing thread feeding between the feed screw blocks 44 and the threaded shaft 41. The threaded shaft 41 can drive the moving chamber 22 to slide axially. The interlocking assembly 5 is operated to separate the guide block 47 from the threaded shaft 41, and the power assembly 6 is activated to move the feed screw blocks 44 away from the threaded shaft 41. At the same time, the compression block 43 is brought into close contact with the threaded shaft 41, rotating the drive ring seat. The compression block 43 drives the threaded shaft 41 to rotate in place, thereby driving the moving chamber 22 to rotate in place.

[0057] 8 and 9, the power assembly 6 includes a drive 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 housing cavity 68 is provided within the drive ring seat 42. The drive ring gear 61 is rotatably connected to the wall of the housing cavity 68 and shares a central axis with the drive ring seat 42. The drive 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 wall of the housing cavity 68. The rotary drive part 69 includes a second electric motor 691 fixedly connected to the wall of the housing cavity 68 and a rotary drive gear 692 fixedly connected to the second electric motor 691. The rotary drive gear 692 meshes with the inner ring wall of the drive 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, and the first gears 62 and the feed screw blocks 44 are provided in one-to-one correspondence, and the second gears 63 and the compression blocks 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 screw block 64 is fixedly connected to one end of the lead screw block 44 that does not extend into the receiving cavity 68, and the first screw block 64 is threadedly connected to the inner peripheral wall of the first thread insert 66. The second thread insert 67 is connected to the second gear 63 via a second bevel gear set 8, which includes a second drive bevel gear 82 coaxially connected to the second gear 63 and a second reverse bevel gear 81 fixedly connected to the second thread insert 67, and the second reverse bevel gear 81 meshes with the second drive bevel gear 82. The second screw block 65 is fixedly connected to one end extending into the accommodating cavity 68 of the compression block 43, and the second screw block 65 is threadedly connected to the inner peripheral wall of the second thread insert 67.

[0059] The second electric motor 691 is driven to rotate the drive ring gear 61 in the forward direction, thereby rotating the first gear 62 and the second gear 63. The first gear 62 drives the rotation of the first threaded insert 66 via the first bevel gear set 7, and the second gear 63 drives the rotation of the second threaded insert 67 via the second bevel gear set 8. The first threaded insert 66 and the first screw block 64 generate yarn feed, and the second threaded 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. Therefore, the compression block 43 and the feed screw block 44 slide in opposite directions, and 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 drive ring gear 61 is rotated in the reverse direction, 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 formed on the upper surface of the support block 51. The groove wall of the first arc trough 55 is smooth and fits against the threaded shaft 41. The press block 52 is slidably mounted on the support block 51 in a vertical direction and is located above the threaded shaft 41. The press block 52 has a second arc trough 55 on the lower surface. The trough 56 is drilled, 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 top surface of the support block 51, and 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, and 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 threadedly connected to the lifting screw 53. The power gear 59 engages with the outer ring wall of the lifting screw ring 54. The third electric motor 58 is driven, and the power gear 59 engages with the lifting screw ring 54, the lifting screw 53 and the lifting screw ring 54 generate yarn feeding, the press block 52 moves downward, and the second arc trough 56 comes into close contact with the threaded shaft 41, thereby fixing the guide block 47 relative to the threaded shaft 41.

[0061] The working principle of Example 3 is as follows: initially, the moving chamber 22 is separated from the fixed chamber 21, and the carrier cover 26 is in a fixed position with most of it extending from the guide sleeve 25. According to the decellularization parameters of Example 1, the packaging bag is placed into the carrier cover 26. The second electric motor 691 is driven, the compression block 43 is separated from the threaded shaft 41, the feed screw block 44 is brought into contact with the threaded shaft 41, and the third electric motor 58 is activated, the press block 52 moves downward, the second arc trough 56 is brought into close contact with the threaded shaft 41, driving the drive element 46, the drive ring seat 42 rotates, and the threaded shaft 41 drives the moving chamber 22 to move axially toward the fixed chamber 21. The moving chamber 22 is inserted into the fixed chamber 21. During this process, the guide ball 29 rolls along the linear arc groove 262.

[0062] After the movable chamber 22 reaches the predetermined position, the second motor 691 is driven, causing the compression block 43 to contact the threaded shaft 41, the feed screw block 44 to separate from the threaded shaft 41, and the driving element 46 is activated, causing the driving ring seat 42 to rotate, and the threaded shaft 41 to drive the movable chamber 22 to rotate in place. At this time, the guide ball 29 enters the spiral arc groove 261, and the spherical wall of the guide ball 29 presses against the groove wall of the spiral arc groove 261 and rolls along the spiral arc groove 261. The carrier cover 26 moves axially toward the guide sleeve 25. After the carrier cover 26 reaches the predetermined position, the guide ball 29 moves into the linear arc groove 262, which then activates the driving element 46 to position the movable chamber 22 at the appropriate position in the fixed chamber 21 and completes the loading of the product.

[0063] When the water pump 391 is started, the fluid medium is introduced into the fixed chamber 21. After the bubbles are removed from 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, and the movable chamber 22 is separated from the fixed chamber 21. During the movement of the movable 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 tested, 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 specimen: The test specimen is immersed in a protease 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 specimen is calculated from the mass of the test specimen and the amount of DNA.

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

[0068] (2) Measurement of DNA content of 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 rate) = (mass of dried test piece) / (mass of test piece).

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

[0071] 2. Calculate the decellularized DNA ratio.

[0072] According to the solutions in steps (1) and (2), the DNA content of the dried specimen of undecellularized tissue is detected. The calculation formula for 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 undecellularized tissue).

[0074] 2. Cell migration test L929 cells (mouse fibroblasts) were cultured in MEM medium at 37°C for 24 hours to perform starvation. 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 placed on the insert (1.0 x 10 5 The cells were seeded at 1000 x 1000 cells / well and cultured at 37°C for 3 hours, and the number of cells that migrated to the bottom of the insert was counted. The cell count was performed 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 cells derived from rat adrenal medulla) were cultured in collagen-coated 24-well culture dishes (1.0 x 10) with RPMI medium containing 10% horse serum and 5% fetal bovine serum (FBS). 4 Cells were seeded at 1000 cells / well and cultured at 37°C for 24 hours. The medium was then switched to RPMI medium containing 0.1% horse serum, and a cell culture insert (pore size 8 μm) was placed in each well. 200 μL of saline containing 5% decellularized tissue was added to the insert, and the culture was continued for 24 hours with or without the addition of 50 ng of nerve growth factor. The cells were observed using a phase-contrast microscope, and the effect of neurite outgrowth was estimated according to the following criteria.

[0076] 2. Decellularization Stability Detection Twenty decellularization treatments were performed according to the decellularization steps of Examples 1 and 2, and then the decellularized DNA ratio was calculated and a cell migration test and a neurite outgrowth test were performed on 20 sets of decellularized tissues of Example 1 and 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 were determined, and the performance deviation value = (|(n performance of Example)-(performance of Example 1)|) / (performance of Example 1)) was calculated, and the average value was finally recorded.

[0077] The detection results are shown in Table 1. Decellularized tissue performance detection table JPEG0007727862000001.jpg85146

[0078] First, as can be seen from the detection results, whether it is blood vessels or bony pig bone marrow, the decellularized tissue obtained in this application has low cytotoxicity and excellent cell induction differentiation effects. Furthermore, after performing the same decellularization process 20 times in both Example 1 and Example 2, the performance deviation of the final decellularized tissue did 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 examples as necessary without making any creative contribution, but all such amendments within the scope of the claims of the present application shall be protected by the Patent Law. [Explanation of symbols]

[0080] 1 base 11 First Base Block 2. Pressure chamber 21 Fixed Chamber 211 Exhaust pipe 212 Second control valve 213 Drainage tube 214 Third control valve 22 Mobile Chamber 221 Second Base Block 23 First Seal 24 Link 1 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 Third 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 First 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 Relief groove 58 Third Electric 59 Power Gear 6 Power Assembly 61 Drive ring gear 62 First Gear 63 2nd Gear 64 First screw block 65 Second screw block 66 First thread insert 67 Second thread insert 68 Containment Cavity 69 Rotational drive parts 691 Second Electric 692 Rotary Drive Gear 7 No. 1 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: Removing tissue of non-human origin; 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, and then the packaging bag is placed in a fluid medium and a water purification pressure is applied to the packaging bag, the treatment solution containing a nuclease and a buffer solution as components; and post-washing the tissue after the high pressure treatment, In the high-pressure treatment step, the packaging bag is placed in a high-voltage device and subjected to high-pressure treatment. The high-voltage device comprises 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 end in the axial direction. The outer periphery of the moving chamber (22) is a peripheral wall that opens at one end in the axial direction. The opening of the moving chamber (22) is The end of the movable chamber (22) is inserted into the fixed chamber (21), and the outer peripheral wall of the movable chamber (22) and the inner peripheral wall of the fixed chamber (21) are in contact with each other. 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 in contact with 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 A carrier cover (26) is slidably mounted within the guide sleeve (25), and 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 in the outer peripheral wall of the carrier cover (26), and a linear arc groove (262) is further formed in 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), and the linear arc groove (262) and the spiral arc groove The carrier cover (26) is in communication with the guide sleeve (25), one end of the carrier cover (26) extending from the guide sleeve (25) is 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, one end of the guide ball (29) that is away from the moving chamber (22) is inserted into the linear arc groove (262) or the spiral arc groove (261), and the spherical wall of the guide ball (29) isThe water injection assembly (3) is in contact with the groove wall of the straight 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 in communication with each other. A method for processing biological tissue for transplantation.

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

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

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

2. The method for treating 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 treating biological tissue for transplantation according to claim 3.

5. A magnetic layer (27) is provided on both the inner peripheral wall of the guide sleeve (25) and the outer peripheral wall of the carrier cover (26).

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

6. The drive assembly (4) comprises a threaded shaft (41), a drive ring seat (42), a compression block (43) and a lead 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 movement of the threaded shaft (41) is One end of the threaded shaft (41) away from the chamber (22) passes through the inner ring wall of the drive 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 drive ring seat (42). At least three of the feed screw blocks (44) 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 at least three feed screw blocks (44) are uniformly distributed in the circumferential direction of the drive ring seat (42), and the feed screw blocks (44) are , one end of the threaded shaft (41) is abutted against the threaded shaft (41) and screw-connected thereto, at least three of the compression blocks (43) 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 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), and at least The three compression blocks (43) and the at least three feed screw blocks (44) slide in opposite directions to each other, and a power assembly (6) for driving the slide movements of the feed screw blocks (44) and the compression blocks (43) is provided on the drive ring seat (42). A guide block (47) is provided on the base (1) so as to be slideable 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).

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

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

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

8. 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 facing the open end. The first pipe trough (361) and the second pipe trough (362) are both passed through the attachment groove (36) and are connected to the front end of the moving chamber (22). The water inlet 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 mounted on the base (1). One end of the water inlet pipe head (32) located within the mounting groove (36) is sealed. A water 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 pipe (39) is connected to the water pump (39). A first control valve (392) is provided at one end of the pipe trough (361) near the fixed chamber (21), one end of the first pipe trough (361) that passes through the mounting groove (36) and one end of the second pipe trough (362) that passes through the mounting groove (36) are provided side by side on the groove wall of the mounting groove (36), a water 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 on 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 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 water passage (351) is provided within the spray ball head (35). The water passage (351) isThe communicating pipe (34) penetrates the spherical wall of the spray ball head (35) extending to the fixed chamber (21), and one end of the communicating 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).

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

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