Method for preparing animal-derived biomaterial and processing device thereof

Abstract

The application provides a preparation method and a processing device of animal-derived biomaterials, which are prepared by combined use of alcohol solution, neutral salt solution, glutaraldehyde solution, detoxification solution and low-concentration sterilization solution. The corresponding processing device comprises a support frame, a honeycomb base, a fixing frame, a fixed container and a peristaltic pump, and the support frame and the honeycomb base can be selected or adjusted in shape and size according to the shape and size of the animal-derived biomaterials. The preparation method and the processing device of the animal-derived biomaterials provided by the application can simultaneously and batch-process multiple pieces of animal-derived biomaterials, significantly improve the processing efficiency, and the prepared animal-derived biomaterials have the advantages of complete cell removal and antigenicity, complete retention of extracellular matrix components, low possibility of calcification and low cytotoxicity.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medical devices, and particularly relates to a preparation method of animal-derived biomaterial and a treatment device thereof. BACKGROUND

[0002] Biological source materials composed of mammalian extracellular matrix are widely used in surgical wound repair, tissue reconstruction and scaffold materials for tissue engineering due to their good biocompatibility. At present, the main components of medical devices such as "patch" and "biological valve" used in clinical practice are mostly derived from the pericardial tissue of pigs, cattle, horses and other animals. Such animal-derived biomaterials need to be treated by decellularization, anti-calcification, multiple rinsing and other operations before being used in the human body. At the same time, due to the serious late calcification and high functional failure rate of animal-derived biomaterials after implantation in the human body, the field pays more attention to the anti-calcification and modification methods of animal-derived biomaterials, and ignores the operation process itself. Therefore, the present application not only provides a preparation method of animal-derived biomaterials, but also provides a treatment device to standardize and simplify the operation process of animal-derived biomaterials.

[0003] There are many kinds of existing anti-calcification methods, and the effects are not accurate. Common methods mainly include surfactants, competitive inhibitors and new cross-linking agents. Surfactants use their washing effect to wash out and reduce phospholipids in the tissue, so that calcium avoids combining with phosphoric acid produced by phospholipids under the action of alkaline phosphatase, thereby inhibiting the formation of calcium salt. Competitive inhibitors use metal ions such as Mg 2+ , Al 3+ , Fe 2+ , etc. to occupy the calcium binding sites at the carboxyl end of collagen molecules or use amino compounds to neutralize aldehyde groups to competitively inhibit the calcium binding sites to achieve the purpose of inhibiting the formation of calcium salt. New cross-linking agents use compounds such as polyepoxy compounds to react with free amino acids of collagen molecules to form covalent bonds to play a cross-linking role.

[0004] Chinese patent document CN202110351059.5 provides a composition for removing phospholipids and cell fragments with alcohols and alcohol ethers, and a method for removing phospholipids and cell fragments on biological tissues. This method only uses alcohols and alcohol ethers to remove phospholipids and cell fragments, and then directly sterilizes the solidified product. Without any elution or other treatment, the cytotoxicity caused by the residues of alcohol and glutaraldehyde after solidification is not considered, which is not conducive to implantation in the body.

[0005] Chinese patent document CN201811391606.7 provides a method for anti-calcification treatment of biological materials, comprising: removing fat; decellularization treatment; glutaraldehyde treatment; soaking in tannic acid solution; soaking in 2,3-butanediol solution; and preserving in glutaraldehyde preservation solution. The decellularization method is not described in detail in this method, tannic acid can stabilize elastic fibers, mucopolysaccharides and other substances, and reduce calcification caused by degradation of the substances, but its cytotoxicity is relatively large, 2,3-butanediol can reduce the cytotoxicity of residual aldehyde groups by combining with aldehyde groups, but the treatment of residual tannic acid toxicity is not considered.

[0006] Chinese patent document CN202021545403.1 provides a tissue fixation treatment device, which is vertically hung by a hook, and a heart pericardium is fixed by a clamp, and the length of the fixing clamp is adjusted to completely unfold the heart pericardium. The problems of this device are: the hook is vertically hung, which cannot fix the position and has low space utilization; the surface of the animal heart pericardium is smooth, and the clamp is easily slipped and easily damages the tissue; the length of the fixing clamp needs to be adjusted for unfolding, which is complex and easily causes uneven stress on the fixed heart pericardium. SUMMARY

[0007] The purpose of the present application is to overcome the deficiencies of the prior art, and to provide not only a preparation method of animal-derived biological materials, but also a device matched with the treatment process.

[0008] The preparation method of animal-derived biological materials provided by the present application comprises the following steps:

[0009] Step 1: removing the fat and loose connective tissue adhered to the animal-derived biological materials;

[0010] Step 2: cleaning the animal-derived biological materials with a neutral salt solution to remove surface fat and free fibers, fixing the animal-derived biological materials on a support frame and clamping them into a fixing frame of a fixing container for corresponding treatment;

[0011] Step 3: adding an alcohol solution to decellularize the animal-derived biological materials;

[0012] Step 4: draining the solution in step 3, adding a neutral salt solution to flush the animal-derived biological materials;

[0013] Step 5: adding a glutaraldehyde solution to solidify the animal-derived biological materials, and preferably replacing the solution every certain period of time, a total of 1-4 times of replacement;

[0014] Step 6: draining the solution in step 5, adding a detoxification solution to reduce the toxicity of glutaraldehyde to the animal-derived biological materials;

[0015] Step 7: draining the solution in step 6, adding a neutral salt solution, and rinsing the animal-derived biomaterial;

[0016] Step 8: draining the solution in step 7, taking the support frame off the fixing frame, cutting the animal-derived biomaterial, and placing it in a neutral salt solution for storage;

[0017] Step 9: sterilizing the animal-derived biomaterial in a sterilization solution, and storing it after sterilization.

[0018] According to the present application, the removal in each of the above steps refers to removing as much as possible the fat, connective tissue, or free fibers, etc. that are desired to be removed, for example, after the removal step, the remaining fat, connective tissue, or free fibers are not more than 5%, preferably not more than 4%, or 3%, more preferably not more than 2% or 1%, still more preferably not more than 0.5%. The most preferred removal step is until no obvious visible oil particles or lint-like tissue adhesion can be seen. The draining refers to draining as much as possible of the target solution, when the next step is rinsing, the animal-derived biomaterial is considered to have been drained when it is completely separated from the solution, preferably the remaining solution after the solution is drained is not more than 5% of the solution before draining, or 4%, or 3%, more preferably not more than 2% or 1%, still more preferably not more than 0.5%, and most preferably there is no residual solution; when the next step is a step that puts the solution in a mixed state, such as detoxification, etc., preferably the remaining solution after the solution is drained is not more than 5% of the solution before draining, considered to have been drained, preferably not more than 4%, or 3%, more preferably not more than 2% or 1%, still more preferably not more than 0.5%, and most preferably there is no residual solution.

[0019] According to the preparation method of the animal-derived biomaterial of the present application, the animal-derived biomaterial in step 1 is the pericardium taken from an animal heart, and the animal is preferably a pig, a cow, or a horse.

[0020] According to the preparation method of the animal-derived biomaterial of the present application, the neutral salt solution in steps 2, 4, 7, and 8 is a mixed solution of phosphate and sodium chloride with a pH value of 7.0-8.0.

[0021] According to the preparation method of the animal-derived biomaterial of the present application, the temperature conditions in steps 2, 3, 4, 5, and 7 are all 10-30°C.

[0022] According to the preparation method of the animal-derived biomaterial of the present application, in steps 3, 4, 5, 6, and 7, the peristaltic pump is turned on, and the speed of the peristaltic pump is adjusted to 400-550 rpm.

[0023] The alcohol in the alcohol solution in step 3 of the preparation method of the animal-derived biomaterial according to the present application is methanol, ethanol, isopropyl alcohol or propanol, preferably ethanol.

[0024] The alcohol solution in step 3 of the preparation method of the animal-derived biomaterial according to the present application has a concentration of 50%-100%, preferably a concentration of 70%, and a treatment time of 48-168 hours.

[0025] The glutaraldehyde solution in step 5 of the preparation method of the animal-derived biomaterial according to the present application has a pH value of 7.0-8.0, a concentration of 0.5%-1% glutaraldehyde solution, and preferably a solution concentration of 0.625%.

[0026] The detoxification solution in step 6 of the preparation method of the animal-derived biomaterial according to the present application is a 4-hydroxyethylpiperazine ethanesulfonic acid solution with a pH value of 5.0-6.0, a treatment temperature condition of 35°C-40°C, and a treatment time of 24-72 hours.

[0027] The preservation temperature condition in step 8 of the preparation method of the animal-derived biomaterial according to the present application is 2°C-8°C, and the preservation time is usually 24 hours to 14 days, and the longest preservation time is 28 days.

[0028] The sterilization solution in step 9 of the preparation method of the animal-derived biomaterial according to the present application is a glutaraldehyde alcohol solution with a pH value of 7.0-8.0 and a concentration of 0.3%-1%, preferably a concentration of 0.4%-0.6%, a treatment temperature condition of 37°C-45°C, a treatment time of 24-48 hours, and a storage temperature condition of 2°C-8°C.

[0029] The alcohol in the glutaraldehyde alcohol solution in the preparation method of the animal-derived biomaterial according to the present application is methanol, ethanol, isopropyl alcohol or propanol, preferably isopropyl alcohol, and the alcohol concentration is 10%-40%, preferably 15%-25%.

[0030] In steps 3, 5 and 6 of the preparation method of the animal-derived biomaterial according to the present application, the peristaltic pump is turned on to keep the solution in a mixed state.

[0031] In steps 4 and 7 of the preparation method of the animal-derived biomaterial according to the present application, the peristaltic pump is turned on to keep the solution in a flowing state, and the flushing time is 10-60 minutes.

[0032] In step 5 of the preparation method of the animal-derived biomaterial according to the present application, the interval time for changing the solution is 24 hours, 48 hours or 72 hours.

[0033] The animal-derived biomaterial processing device provided by the application comprises a support frame, a honeycomb base, a fixing frame, a fixed container and a peristaltic pump; the support frame comprises an inner ring, an outer ring and a buckle; the honeycomb base has a protruding part for supporting animal-derived biomaterials, and the surface of the honeycomb base has a honeycomb hole structure; the shape of the inner ring matches the shape of the protruding part, and the inner circumference of the inner ring matches the outer circumference of the protruding part, so that the inner ring can be sleeved on the outer sidewall of the protruding part; the shape of the outer ring matches the shape of the inner ring, and the inner circumference of the outer ring matches the outer circumference of the inner ring, so that the outer ring and the inner ring can be tightly sleeved together; the fixing frame is detachably installed in the fixed container, and the fixing frame is provided with a clamping groove; the outer ring is provided with a buckle, the buckle can close the outer ring, and the support frame loaded with animal-derived biomaterials is installed on the fixing frame through the cooperation of the buckle and the clamping groove; the peristaltic pump is used in cooperation with the high-level faucet and the low-level faucet on the fixed container, and can realize the solution adding, draining, mixing, liquid changing or flowing operation in the fixed container.

[0034] According to the animal-derived biomaterial processing device, the support frame and the honeycomb base are selected according to the different shapes and sizes of animal-derived biomaterials.

[0035] According to the animal-derived biomaterial processing device, the inner ring and the outer ring of the support frame, and the protruding part of the honeycomb base are made into corresponding shapes and sizes according to the different shapes and sizes of animal-derived biomaterials.

[0036] According to the animal-derived biomaterial processing device, the fixed container is selected according to the number and size of animal-derived biomaterials.

[0037] According to the animal-derived biomaterial processing device, the fixing frame is selected according to the size of the fixed container.

[0038] According to the animal-derived biomaterial processing device, the clamping groove of the fixing frame is determined according to the number of animal-derived biomaterials.

[0039] The animal-derived biomaterial processing device according to the present application, in the solution adding, draining, mixing, liquid changing or flowing operation in the fixed container, the solution adding is low faucet off, high faucet on, the solution only goes in, stops operation after the solution covers the animal-derived biomaterial; the draining is high faucet off, low faucet on, the solution only goes out; the mixing is low faucet out, high faucet in, the existing solution in the container is recycled; the liquid changing is to drain the original solution first and then add new solution; the flowing is to add new solution after the operation is completed, high faucet in, low faucet out, and the flowing state is maintained.

[0040] Technical effects of the present application:

[0041] The method for preparing animal-derived biomaterials and the corresponding processing device with simple structure and easy operation can be used repeatedly, and the two are used together to simultaneously batch process multiple pieces of animal-derived biomaterials, significantly improve the processing efficiency, efficiently and completely remove the cell components and antigenicity causing rejection reaction in the animal-derived biomaterials, retain the extracellular matrix components, and maximally reduce the possibility of calcification and the cytotoxicity, and are widely used in the field of animal-derived biomaterial preparation.

[0042] The preparation method of the present application can effectively and completely destroy cells, remove phospholipids and cell fragments through dehydration, and also has the effects of anti-calcification and depyrogenation by prolonging the processing time of the alcohol solution; the flushing of the neutral salt solution can remove alcohol residues on the animal-derived biomaterials; the glutaraldehyde solidification connects collagen molecules into stable cross-linked structures, thereby covering and closing the active genes in the protein molecules, making them lose antigenicity, and the multiple liquid changes in the solidification process can prevent the concentration of glutaraldehyde in the solution from decreasing due to the consumption of glutaraldehyde by the materials to form polyglutaraldehyde cross-linking; the treatment of detoxification solution after solidification and the multiple flushing of the neutral salt solution can also effectively reduce the residual glutaraldehyde on the surface and gaps of the materials, thereby reducing the possibility of calcification with the proteins in the bound tissues and the chance of toxic reaction in the patient's body; the reduction of the concentration of glutaraldehyde in the glutaraldehyde alcohol-containing sterilization solution can make the residual degree of glutaraldehyde lower after rinsing before implantation under the premise that the sterilization solution does not damage the tissue structure and can effectively sterilize.

[0043] The processing device of the present application, in which the mutual cooperation of each part of the support frame enables each piece of animal-derived biomaterial to be fixed flat, and this fixing method has the advantages of no damage to the material, uniform stress, not easy to slip off, simple operation, etc.; the cooperation of the buckle and the slot not only fixes the position of each piece of material, but also greatly improves the space utilization; the cooperation of the peristaltic pump and the high and low faucets on the fixed container not only makes the rotation speed (i.e. the flow rate of the solution) adjustable, but also realizes multiple operation states of solution adding, draining, mixing and flowing. BRIEF DESCRIPTION OF DRAWINGS

[0044] Figure 1 This is a schematic diagram of the support frame and honeycomb base according to an embodiment of the present invention;

[0045] Figure 2 This is a schematic diagram of the installation of the fixed container and support frame according to an embodiment of the present invention. Detailed Implementation

[0046] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that the embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present invention. Furthermore, it should be understood that after reading the disclosure of this invention, those skilled in the art can make various modifications or alterations to the invention, and these equivalent forms also fall within the scope of protection defined by this invention.

[0047] The embodiments of the method for preparing animal-derived biomaterials and the processing apparatus provided by the present invention include the following steps:

[0048] Step 1: Remove the pericardium from the heart of animals such as pigs, cows, and horses, and carefully remove any adhering fat and loose connective tissue.

[0049] Step 2: Cleanse the pericardium with a neutral salt solution of pH 7.2 to remove surface fat and free fibers. Depending on the size and shape of the pericardium, select a support frame and honeycomb base of appropriate size and shape to simultaneously process pericardiums of different sizes and shapes. For example... Figure 1 As shown, the inner ring 1.2 is fitted onto the protrusion 1.1 of the honeycomb base, and the honeycomb base is placed in a neutral saline solution. The pericardium is then immersed in the neutral saline solution, allowing it to float freely and gently spread on the surfaces of the inner ring 1.2 and the protrusion 1.1 of the honeycomb base. The honeycomb base has a honeycomb-like structure 1.4, which prevents the pericardium from adhering to the honeycomb base and becoming difficult to remove. The pericardium is fixed with the outer ring 1.3 and the clip 1.5. The outer ring 1.3 has two extended ends at its port, and the clip 1.5 can close the outer ring 1.3 by restraining the two ends at its port. Then, the support frame carrying the pericardium is installed into the slot 2.1 of the fixing frame 2 through the clip 1.5. The clip 1.5 and the slot 2.1 are fixed together. Subsequently, the fixing frame 2 is placed in the prepared fixing container 3.

[0050] Step 3: Add 70% ethanol solution, cover with the top cap, turn on the peristaltic pump 4, turn on the low-position tap 3.3 and the high-position tap on the fixed container 3, adjust the speed to 500 rpm to mix the solution, and treat at 23℃ for 48 hours.

[0051] Step 4: open the low-positioned faucet 3.3 on the fixed container 3 to drain the alcohol solution, add the neutral salt solution with pH value of 7.2, open the peristaltic pump 4 at 23℃, adjust the rotating speed to 500 rpm, keep the solution flowing for 45 minutes for flushing treatment;

[0052] Step 5: add the glutaraldehyde solution with pH value of 7.4 and concentration of 0.625% at 23℃, cover the pericardium with the top cover, open the peristaltic pump 4 at 500 rpm, and adjust the rotating speed to 500 rpm, cooperate with the high-positioned faucet and the low-positioned faucet 3.3 on the fixed container, mix the solution thoroughly, change the solution every day during the solidification process, and complete the change for a total of 2 times;

[0053] Step 6: open the low-positioned faucet 3.3 to drain the solidification solution after the completion of Step 5, add the 4-hydroxyethylpiperazine ethanesulfonic acid solution at 37℃, cover the pericardium with the top cover, open the peristaltic pump 4 at 500 rpm, adjust the rotating speed to 500 rpm, cooperate with the high-positioned faucet and the low-positioned faucet 3.3 on the fixed container, mix the solution thoroughly, and perform the glutaraldehyde toxicity reduction treatment on the pericardium for 48 hours. In the initial stage of adding the 4-hydroxyethylpiperazine ethanesulfonic acid solution, the low-positioned faucet 3.3 can be kept open for a period of time to completely drain the solidification solution;

[0054] Step 7: open the low-positioned faucet 3.3 to drain the detoxification solution after the completion of Step 6, add the neutral salt solution with pH value of 7.2, open the peristaltic pump 4 at 23℃, adjust the rotating speed to 500 rpm, keep the solution flowing for 45 minutes for flushing treatment;

[0055] Step 8: open the low-positioned faucet 3.3 to drain the neutral salt solution after the completion of Step 7, remove the support frame from the clamping groove 2.1 of the fixed frame 2, loosen the buckle 1.5, remove the inner ring 1.2, the outer ring 1.3, and the pericardium, cut the flushed pericardium according to the shape of the inner ring 1.2 with dissecting scissors, and store it in the neutral salt solution with pH value of 7.2. The storage temperature is 2℃-8℃, and the storage time is usually 24 hours to 14 days, and the longest storage time is 28 days;

[0056] Step 9: place the pericardium in Step 8 in the glutaraldehyde alcohol solution with pH value of 7.4 and concentration of 0.3%-1%, preferably 0.4%-0.6%, and more preferably 0.5%. The alcohol in the solution is isopropyl alcohol with a concentration of 20%, sterilize it at 41℃ for 48 hours, and store it at 2℃-8℃ after sterilization for future use.

[0057] According to the present application, the removal in each step above means that the fat, connective tissue or free fiber etc. to be removed is removed as much as possible, for example, after the removal step, the remaining fat, connective tissue or free fiber is not more than 5%, preferably not more than 4%, or 3%, more preferably not more than 2% or 1%, still more preferably, not more than 0.5%. The most preferred removal step is until no obvious visible oil particles or floc-like tissue adhesion can be seen. The draining means that the target solution is drained as much as possible, when the next step is washing, the solution is considered to be drained when the animal-derived biological material is completely separated from the solution, preferably the remaining solution after the solution is drained is not more than 5% of the solution before draining, or 4%, or 3%, more preferably not more than 2% or 1%, still more preferably, not more than 0.5%, most preferably there is no residual solution; when the next step is a step such as detoxification etc. to make the solution in a mixed state, preferably the remaining solution after the solution is drained is not more than 5% of the solution before draining, considered to be drained, preferably not more than 4%, or 3%, more preferably not more than 2% or 1%, still more preferably, not more than 0.5%, most preferably there is no residual solution.

[0058] According to another embodiment of the present application, when the pericardium removed from the heart of a pig, a cow, a horse or the like is relatively smooth, only one step can be used to remove the surface fat and free fiber, i.e. step 1 of manually removing the adhered fat and loose connective tissue is omitted, and only a neutral salt solution is used for washing.

Claims

1. An apparatus for processing an animal-derived biological material, characterized by comprising: The processing device comprises a support frame, a honeycomb base, a fixing frame, a fixed container and a peristaltic pump; the support frame comprises an inner ring, an outer ring and a buckle; the honeycomb base has a protruding part for supporting animal-derived biological materials, and the surface of the honeycomb base has a honeycomb hole structure; the shape of the inner ring matches that of the protruding part, and the inner circumference of the inner ring matches the outer circumference of the protruding part, so that the inner ring can be sleeved on the outer sidewall of the protruding part; the shape of the outer ring matches that of the inner ring, and the inner circumference of the outer ring matches the outer circumference of the inner ring, so that the outer ring and the inner ring can be tightly sleeved together; the fixing frame is detachably installed in the fixed container, the fixing frame is provided with a clamping groove, the outer ring is provided with a buckle, the buckle can close the outer ring, and the support frame loaded with animal-derived biological materials is installed on the fixing frame through the cooperation of the buckle and the clamping groove; the peristaltic pump is used in cooperation with the high-position faucet and the low-position faucet on the fixed container, and can realize the adding, draining, mixing, liquid changing or flowing operation of the solution in the fixed container. The support frame and the honeycomb base are selected according to the shape and size of the animal-derived biological materials.

2. The animal-derived biological material processing apparatus according to claim 1, wherein The inner ring and the outer ring of the support frame and the protruding part of the honeycomb base are made into corresponding shapes and sizes according to the shape and size of the animal-derived biological materials.

3. The animal-derived biological material processing apparatus according to claim 1, wherein The fixed container is selected according to the quantity and size of the animal-derived biological materials.

4. The animal-derived biological material processing apparatus according to claim 1, wherein The fixing frame is selected according to the size of the fixed container.

5. The animal-derived biological material processing apparatus according to claim 1, wherein The interval distance of the clamping groove of the fixing frame is determined according to the quantity of the animal-derived biological materials.

6. The animal-derived biological material processing apparatus according to claim 1, wherein In the adding, draining, mixing, liquid changing or flowing operation of the solution in the fixed container, the adding of the solution is that the low-position faucet is closed, the high-position faucet is opened, the solution only enters but does not exit, and the operation is stopped after the solution covers the animal-derived biological materials; the draining is that the high-position faucet is closed, the low-position faucet is opened, and the solution only exits but does not enter; the mixing is that the low-position faucet discharges water, the high-position faucet discharges water, and the existing solution in the container is recycled; the liquid changing is that the original solution is drained first and then the new solution is added; and the flowing is that after the adding operation is completed, the high-position faucet discharges water, the low-position faucet discharges water, and the flowing state is maintained by continuously adding the new solution.

7. The animal-derived biological material processing apparatus according to any one of claims 1 to 6, characterized by, The processing device is prepared by using the processing device of claim 1, comprising the following steps:

8. A method for preparing an animal-derived biomaterial, characterized by, Step 1: removing the fat and loose connective tissue adhered to the animal-derived biological materials; Step 2: cleaning the animal-derived biological materials with a neutral salt solution, removing the surface fat and free fibers, fixing the animal-derived biological materials on the support frame and clamping the support frame into the fixing frame of the fixed container for corresponding processing; Step 3: adding an alcohol solution to the animal-derived biological materials for decellularization treatment, and the treatment time is 48-168 hours; Step 4: draining the solution in step 3, adding a neutral salt solution to the animal-derived biological materials for washing treatment; Step 5: adding a glutaraldehyde solution to the animal-derived biological materials for solidification, and performing liquid changing every certain period of time, and the total number of liquid changing is 1-4 times. ​ Step 6: drain the solution in step 5, add detoxification solution to reduce glutaraldehyde toxicity of the animal-derived biological material; Step 7: drain the solution in step 6, add a neutral salt solution to flush the animal-derived biological material; Step 8: drain the solution in step 7, remove the support frame from the fixing frame, cut the animal-derived biological material, and store it in a neutral salt solution; Step 9: sterilize the animal-derived biological material in a sterilization solution, and store it after sterilization is completed.

9. The method for preparing animal-derived biomaterials according to claim 8, characterized in that, The animal-derived biological material in step 1 is pericardium taken from the heart of an animal, and the animal is a pig, a cow, or a horse.

10. The method for preparing animal-derived biomaterials according to claim 9, characterized in that, The neutral salt solution in steps 2, 4, 7, and 8 is a mixed solution of phosphate and sodium chloride with a pH value of 7.0-8.

0.

11. The method for preparing animal-derived biomaterials according to claim 9, characterized in that, The temperature conditions in steps 2, 3, 4, 5, and 7 are all 10-30°C.

12. The method of claim 9, wherein the animal-derived biomaterial is prepared by the steps of: In steps 3, 4, 5, 6, and 7, the rotation speed of the peristaltic pump is adjusted to 400-550 rpm.

13. The method for preparing animal-derived biomaterials according to claim 9, characterized in that, The alcohol in the alcohol solution in step 3 is methanol, ethanol, isopropanol, or propanol.

14. The method for preparing animal-derived biomaterials according to claim 9, characterized in that, The concentration of the alcohol solution in step 3 is 50-100%.

15. The method of claim 14, wherein the animal-derived biomaterial is prepared by the steps of: The concentration of the alcohol solution in step 3 is 70%.

16. The method of claim 9, wherein the animal-derived biomaterial is prepared by the steps of: The pH value of the glutaraldehyde solution in step 5 is 7.0-8.0, and the concentration is 0.5-1% glutaraldehyde solution.

17. The method of claim 16, wherein the animal-derived biomaterial is prepared by the steps of: The concentration of the glutaraldehyde solution in step 5 is 0.625%.

18. The method for preparing animal-derived biomaterials according to claim 9, characterized in that, The detoxification solution in step 6 is a 4-hydroxyethylpiperazine ethanesulfonic acid solution with a pH value of 5.0-6.0, the temperature condition is 35-40°C, and the time is 24-72 hours.

19. The method for preparing animal-derived biomaterials according to claim 9, characterized in that, The storage temperature condition in step 8 is 2-8°C.

20. The method of claim 8-19, wherein the animal-derived biomaterial is prepared by a method comprising the steps of: The sterilization solution in step 9 is selected to have a pH value of 7.0-8.0, a solution concentration of 0.3-1%, a temperature condition of 37-45°C, a treatment time of 24-48 hours, and a storage temperature condition of 2-8°C.

21. The method of claim 20, wherein the animal-derived biomaterial is prepared by a method comprising the steps of: The sterilization solution in step 9 is a glutaraldehyde alcohol solution with a concentration of 0.4-0.6%.

22. The method of claim 21, wherein the animal-derived biomaterial is prepared by a method comprising the steps of: The alcohol in the glutaraldehyde alcohol solution is methanol, ethanol, isopropanol, or propanol, and the alcohol concentration is 10-40%.

23. The method for preparing animal-derived biomaterials according to claim 22, characterized in that, The alcohol in the glutaraldehyde alcohol solution is isopropanol, and the alcohol concentration is 15-25%.

24. The method of producing an animal-derived biomaterial according to any one of claims 8 to 19, wherein In steps 3, 5, and 6, the peristaltic pump is turned on to keep the solution mixed.

25. The method of producing an animal-derived biomaterial according to any one of claims 8 to 19, wherein In steps 4 and 7, the peristaltic pump is turned on to keep the solution flowing, and the flushing time is 10-60 minutes.

26. The method of producing an animal-derived biomaterial according to any one of claims 8 to 19, wherein In step 5, the interval time for changing the solution is 24 hours, 48 hours, or 72 hours.

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