Decellularized artificial lymph drainage tube as well as preparation method and application thereof

Abstract

The invention discloses a decellularized artificial lymph drainage tube and a preparation method and application thereof, and the preparation method of the decellularized artificial lymph drainage tube comprises the following steps: taking an animal blood vessel as a raw material, and performing decellularization treatment, cleaning treatment, enhancement treatment, hole forming treatment and drying treatment on the raw material. The decellularized artificial lymph drainage tube prepared by the preparation method can solve the problems of long consumed time and difficult operation of the existing lymphatic vessel-vein anastomosis operation, can drain the lymph at the swelling part of a patient to the vein system in time after implantation, and can quickly relieve the swelling symptom. The decellularized artificial lymph drainage tube supports ingrowth, migration and proliferation of autologous cells and is good in biocompatibility, the degradation time can be adjusted to be matched with the reconstruction time of an autologous lymphatic reflux system, and long-term patency is guaranteed.

Description

Technical Field

[0001] The present application relates to the technical field of medical devices and bionic materials, and in particular to a decellularized artificial lymphatic drainage tube and a preparation method and application thereof. Background Art

[0002] Lymphedema is a common chronic disease. Lymphedema is mostly caused by cancer surgery, radiotherapy or infection, which leads to obstruction of lymph circulation and accumulation of tissue fluid. Patients often face problems such as limb swelling, functional impairment and decreased quality of life. The current treatments for lymphedema include conservative treatment (such as pressure therapy, physical therapy) and surgical treatment. Among them, lymphaticovenous anastomosis (LVA) is a minimally invasive surgical method that directly connects tiny (diameter <0.5mm) lymphatic vessels to veins to improve lymphatic return. It is widely used in patients with early lymphedema. The advantages of LVA are small trauma, fast recovery, and significant reduction of swelling within a few days after surgery. However, its main disadvantages include that the success rate is limited by the functional status of the lymphatic vessels, and the requirements for anastomosis equipment and technology are very high.

[0003] Inspired by LVA technology, the drainage end point of an ideal artificial lymphatic drainage tube should be the autologous vein, so that the continuously produced lymph can return in a timely and effective manner. Among them, the autologous vein can be selected from the first-order small branches of the large veins of the upper and lower limbs. These branches usually have valve structures to prevent blood from flowing back into the lymphatic drainage tube. In addition, the selection of appropriate materials is of great significance for the long-term patency, lasting functionality and reconstruction of the autologous lymphatic return system of the artificial lymphatic drainage tube. At present, there have been some reports on the use of synthetic polymer materials to drain lymph (10.1089 / lrb.2018.0042). Artificial lymphatic drainage tubes based on polymer materials usually have good biological inertness and help reduce immune stimulation. However, the hardness of polymer materials is usually large, and they cannot simulate the contraction and relaxation behavior of the human vascular system, and it is difficult to anastomose with human blood vessels; in addition, this type of implanted polymer material is generally non-degradable, cannot support the reconstruction of the lymphatic return system, and there is also a risk of inflammatory hyperplasia-type blockage in the long term. Summary of the invention

[0004] Based on this, it is necessary to provide a decellular artificial lymphatic drainage tube. The decellular artificial lymphatic drainage tube of the present application can solve the problems of the existing lymphatic-venous anastomosis surgery being time-consuming and difficult to operate.

[0005] An embodiment of the present application provides a decellularized artificial lymph drainage tube.

[0006] A method for preparing a decellularized artificial lymph drainage tube comprises the following steps:

[0007] Animal blood vessels are used as raw materials, and the raw materials are subjected to decellularization, cleaning, strengthening, pore making and drying treatments.

[0008] In some embodiments, the animal blood vessels include blood vessels of one or more of pigs, cows, sheep, rabbits, mice, fish, and poultry.

[0009] In some embodiments, the animal blood vessels include one or a combination of arterial blood vessels and venous blood vessels.

[0010] In some embodiments, the decellularization treatment includes one or a combination of alkali treatment, acid treatment, surfactant treatment, and enzyme treatment.

[0011] In some embodiments, the alkali treatment comprises the following steps: placing the raw material in an alkali solution with an alkali concentration of 0.5wt% to 5wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5h to 12h.

[0012] In some embodiments, the acid treatment comprises the following steps: placing the raw material in an acid solution with an acid concentration of 0.1wt% to 0.5wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5h to 12h.

[0013] In some of the embodiments, the surfactant treatment comprises the following steps: placing the raw material in a surfactant solution with a surfactant concentration of 1wt% to 5wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5h to 12h.

[0014] In some embodiments, the enzyme treatment comprises the following steps: placing the raw material in a pancreatic enzyme solution with a pancreatic enzyme concentration of 0.25wt% to 1.0wt%, and performing a decellularization treatment at a temperature of 30°C to 35°C for 4h to 24h.

[0015] In some embodiments, when the decellularization treatment includes a combination of at least two of alkaline treatment, acid treatment, surfactant treatment, and enzyme treatment, a plurality of washing steps are interspersed between the decellularization treatment steps.

[0016] In some embodiments, the drying process includes the following steps: freeze-drying the raw materials after the cleaning process.

[0017] In some embodiments, the reinforcing treatment includes one or a combination of a cross-linking treatment and a surface coating treatment.

[0018] In some embodiments, the cross-linking treatment includes thermal cross-linking, chemical cross-linking, or a combination of both.

[0019] In some embodiments, the thermal crosslinking comprises the following steps: freeze-drying the raw material after the cleaning treatment, and then placing it in a vacuum environment at 60° C. to 120° C. for crosslinking treatment for 1 hour to 24 hours.

[0020] In some of the embodiments, the chemical crosslinking comprises the following steps: placing the cleaned raw material in a chemical crosslinking agent solution with a chemical crosslinking agent concentration of 0.01wt% to 5wt%, and stirring and crosslinking at room temperature or 4°C for 1h to 72h.

[0021] In some embodiments, the chemical crosslinking agent includes one or more combinations of glutaraldehyde, formaldehyde, genipin, butanediol glycidyl ether, divinyl sulfone, oxalic acid dihydrazide, carbodiimide and glycidyl methacrylate.

[0022] In some embodiments, the surface coating treatment includes the following steps: placing the raw material after cleaning in a biocompatible polymer solution with a biocompatible polymer concentration of 0.1wt%~5wt%, soaking for 30s~10min, then taking out the raw material and removing the solvent of the biocompatible polymer.

[0023] In some embodiments, the biocompatible polymer includes one or more combinations of chitosan, cellulose, polylactic acid, silk fibroin, collagen, polyvinyl alcohol, polylactic acid-co-glycolic acid, polymethyl methacrylate, polyamide, polyurethane, and polycaprolactone.

[0024] In some embodiments, the length of the animal blood vessel is 0.5 cm to 50 cm.

[0025] In some embodiments, the outer diameter of the animal blood vessel is 0.1 mm to 6 mm.

[0026] In some embodiments, the animal blood vessel includes one of a straight structure and a bifurcated structure.

[0027] An embodiment of the present application also provides a decellularized artificial lymph drainage tube.

[0028] A decellularized artificial lymph drainage tube is prepared by adopting the above-mentioned preparation method.

[0029] An embodiment of the present application also provides an application of a decellularized artificial lymph drainage tube.

[0030] A decellularized artificial lymphatic drainage tube is used in lymphatic-venous anastomosis, wherein one end of the decellularized artificial lymphatic drainage tube can be anastomosed with an autologous vein or its branch, or with another decellularized artificial lymphatic drainage tube, and the other end of the decellularized artificial lymphatic drainage tube can be anastomosed with an autologous lymphatic vessel, or with another decellularized artificial lymphatic drainage tube or with an opening left in subcutaneous tissue.

[0031] The above-mentioned decellularized artificial lymph drainage tube can solve the problems of time-consuming and difficult operation of existing lymphatic-venous anastomosis surgery, and can drain the lymph from the swollen part of the patient to the venous system in time after implantation, and can quickly relieve the symptoms of swelling. The decellularized artificial lymph drainage tube of the present application supports the growth, migration, and proliferation of autologous cells, has good biocompatibility, and the degradation time can be adjusted to match the reconstruction time of the autologous lymphatic return system to ensure long-term patency. Specifically, the present application has an artificial lymph drainage tube with suitable degradation performance that simulates the elasticity of blood vessels and is of great significance. The decellularized artificial lymph drainage tube based on animal blood vessels as raw materials not only has excellent biocompatibility, but also can retain the elasticity of the blood vessels themselves; its degradation rate can be adjusted according to the degree of enhancement process to match the reconstruction rate of the autologous lymphatic return system, thereby ensuring smooth long-term lymph return. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0033] In order to more completely understand the present application and its beneficial effects, the following description will be made in conjunction with the accompanying drawings. In the following description, the same reference numerals represent the same parts.

[0034] Figure 1 This is a digital photo of the porcine saphenous artery-derived decellularized artificial lymphatic drainage tube described in one embodiment of the present application, wherein Figure 1 A ruler is included to indicate length;

[0035] Figure 2 The outer surface microstructure of the acellular artificial lymph drainage tube derived from the porcine saphenous artery described in one embodiment of the present application;

[0036] Figure 3 The inner surface microstructure of the acellular artificial lymph drainage tube derived from the porcine saphenous artery described in one embodiment of the present application;

[0037] Figure 4This is an H&E stained section image of a porcine saphenous artery before decellularization according to an embodiment of the present application;

[0038] Figure 5 for Figure 4 H&E-stained sections of porcine saphenous artery after decellularization. DETAILED DESCRIPTION

[0039] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0040] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

[0041] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0042] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0043] In the description of the present invention, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, and "above", "below", "within", etc. are understood to include the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0044] In this article, "optionally", "optional", "optional" means optional, that is, it means to be selected from any of the two parallel solutions of "yes" or "no". If there are multiple "optional" in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent. In this application, the descriptions such as "optionally contain" and "optionally include" mean "contain or not contain".

[0045] In the present invention, unless otherwise stated, the sum of the parts of each component in the composition can be 100 parts by weight. Unless otherwise specified, the basis of the percentage (including weight percentage) of the present invention is the total weight of the composition, and "wt%" herein means mass percentage.

[0046] In the present application, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​in the numerical interval is considered to be continuous, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to an integer in the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints, which is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical range disclosed in the present application should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows for a broad range of quantitative intervals such as percentage intervals, ratio intervals, and ratio intervals.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0048] It should be noted that the room temperature herein refers to 18°C ​​to 40°C, preferably, the room temperature refers to 20°C to 30°C.

[0049] The embodiment of the present application provides a decellularized artificial lymphatic drainage tube to improve the problem that some existing polymer drainage materials are usually hard and cannot simulate the contraction and relaxation behavior of the human vascular system; in addition, this type of implanted polymer material is generally non-degradable and cannot support the reconstruction of the lymphatic return system. There is also a risk of inflammatory hyperplasia type blockage in the long term, and further solves the problem that the lymphatic-venous anastomosis surgery in the current technology is time-consuming and difficult to operate. The decellularized artificial lymphatic drainage tube will be described below in conjunction with the accompanying drawings.

[0050] The acellular artificial lymphatic drainage tube provided in the present application is exemplary, see Figure 1 As shown, Figure 1 The schematic diagram of the structure of the decellularized artificial lymphatic drainage tube provided in the embodiment of the present application. The decellularized artificial lymphatic drainage tube of the present application can be used for medical surgery, such as lymphatic-venous anastomosis surgery.

[0051] In order to more clearly illustrate the structure of the decellularized artificial lymph drainage tube, the decellularized artificial lymph drainage tube will be introduced below with reference to the accompanying drawings.

[0052] For example, see Figure 1 As shown, Figure 1 This is a schematic diagram of the decellularized artificial lymph drainage tube provided in an embodiment of the present application.

[0053] It should be noted that, in this article, unless otherwise specified, each reaction step can be carried out in the order described herein, or can be carried out in a non-sequential manner. For example, other steps may be included between each reaction step, and the order of the reaction steps may also be appropriately swapped. This can be determined by the technician based on conventional knowledge and experience. Preferably, the reaction method herein is carried out sequentially.

[0054] A method for preparing a decellularized artificial lymph drainage tube comprises the following steps:

[0055] Animal blood vessels are used as raw materials, and the raw materials are subjected to decellularization, cleaning, strengthening, pore making and drying treatments.

[0056] In some embodiments, the animal blood vessels include blood vessels of one or more of pigs, cows, sheep, rabbits, mice, fish and birds.

[0057] In some embodiments, the animal blood vessel includes one or a combination of arterial blood vessels and venous blood vessels.

[0058] In some embodiments, the decellularization treatment includes one or a combination of alkali treatment, acid treatment, surfactant treatment, and enzyme treatment.

[0059] In some embodiments, the alkali treatment comprises the following steps: placing the raw material in an alkali solution with an alkali concentration of 0.5wt% to 5wt%, and performing a decellularization treatment at room temperature for 0.5h to 12h. Preferably, the concentration of the alkali solution is 0.8wt% to 4wt%. Further preferably, the concentration of the alkali solution is 1.5wt% to 3.5wt%. During the alkali treatment, the value of the decellularization treatment time includes but is not limited to: 0.5h, 1h, 2h, 3h, 4h, 4.5h, 5h, 6h, 7h, 8h, 8.5h, 9h, 10h, 11h, 12h or a range between any two of the foregoing.

[0060] In some embodiments, the alkaline solution includes sodium hydroxide, potassium hydroxide, etc.

[0061] In some embodiments, the acid treatment comprises the following steps: placing the raw material in an acid solution with an acid concentration of 0.1wt% to 0.5wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5h to 12h. Preferably, the concentration of the acid solution is 0.2wt% to 0.4wt%. During the acid treatment, the decellularization time includes but is not limited to: 0.5h, 1h, 2h, 3h, 4h, 4.5h, 5h, 6h, 7h, 8h, 8.5h, 9h, 10h, 11h, 12h or a range between any two of the foregoing.

[0062] In some embodiments, the acid solution includes dilute hydrochloric acid, dilute acetic acid, dilute sulfuric acid, etc.

[0063] In some embodiments, the surfactant treatment includes the following steps: placing the raw material in a surfactant solution with a surfactant concentration of 1wt% to 5wt%, and decellularizing at room temperature or -4°C for 0.5h to 12h. Preferably, the surfactant concentration is 0.8wt% to 4wt%. Further preferably, the surfactant concentration is 1.5wt% to 3.5wt%. During the surfactant treatment, the value of the decellularization time includes but is not limited to: 0.5h, 1h, 2h, 3h, 4h, 4.5h, 5h, 6h, 7h, 8h, 8.5h, 9h, 10h, 11h, 12h or a range between any two of the foregoing.

[0064] In some embodiments, the surfactant includes one or a combination of cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), Tween 20, Tween 80, Triton X-100 and glycerol glucoside.

[0065] In some embodiments, the enzyme treatment comprises the following steps: placing the raw material in a pancreatic enzyme solution with a pancreatic enzyme concentration of 0.25wt% to 1wt%, and decellularizing at a temperature of 30°C to 35°C for 4h to 24h. Preferably, the concentration of the pancreatic enzyme solution is 0.5wt% to 0.8wt%. During the enzyme treatment, the value of the decellularization time includes but is not limited to: 4h, 5h, 6h, 7h, 8h, 8.5h, 9h, 10h, 11h, 12h, 14h, 15h, 18h, 20h, 22h, 24h or a range between any two of the foregoing.

[0066] In some embodiments, the washing process includes the following steps: washing the decellularized raw material in pure water or PBS buffer for 5 to 30 minutes. The washing time includes but is not limited to: 5 minutes, 6 minutes, 8 minutes, 10 minutes, 12 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes or a range between any two of the foregoing.

[0067] In some embodiments, the raw material after decellularization is washed several times or at least once during the washing process.

[0068] In some embodiments, the drying process includes the following steps: freeze-drying the raw materials after the cleaning process.

[0069] In some of the embodiments, the reinforcing treatment includes one or a combination of a cross-linking treatment and a surface coating treatment.

[0070] In some embodiments, the cross-linking treatment includes one or a combination of thermal cross-linking and chemical cross-linking.

[0071] In some embodiments, the thermal crosslinking comprises the following steps: freeze-drying the cleaned raw material, and then placing it in a vacuum environment at 60°C to 120°C for crosslinking for 1h to 24h. The value of the thermal crosslinking time includes but is not limited to: 1h, 6h, 10h, 18h, 24h, or a range between any two of the foregoing.

[0072] In some embodiments, the chemical crosslinking comprises the following steps: placing the cleaned raw material in a chemical crosslinking agent solution with a chemical crosslinking agent concentration of 0.01wt% to 0.5wt%, and stirring and crosslinking for 1h to 72h at room temperature or 4°C. The value of the light crosslinking time includes but is not limited to: 1h, 5h, 10h, 24h, 36h, 48h, 72h, or a range between any two of the foregoing.

[0073] In some embodiments, the chemical cross-linking agent includes one or more combinations of genipin, butanediol glycidyl ether, divinyl sulfone, oxalic acid dihydrazide, carbodiimide, and glycidyl methacrylate.

[0074] In some of the embodiments, the surface coating treatment includes the following steps: placing the cleaned raw material in a biocompatible polymer solution with a biocompatible polymer concentration of 0.1wt%~5wt%, soaking for 30s~10min, then taking out the raw material and removing the solvent of the biocompatible polymer.

[0075] In some embodiments, the biocompatible polymer includes one or more combinations of chitosan, cellulose, polylactic acid, silk fibroin, collagen, polyvinyl alcohol, polylactic acid-co-glycolic acid, polymethyl methacrylate, polyamide, polyurethane and polycaprolactone.

[0076] In some embodiments, the length of the animal blood vessel is 0.5 cm to 50 cm. The length of the animal blood vessel includes but is not limited to: 0.5 cm, 5 cm, 10 cm, 15 cm, 25 cm, 30 cm, 35 cm, 40 cm, 50 cm or a range between any two of the foregoing.

[0077] In some embodiments, the outer diameter of the animal blood vessel is 0.1 mm to 6 mm. The value of the outer diameter of the animal blood vessel includes but is not limited to: 0.1 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm or a range between any two of the foregoing.

[0078] In some embodiments, the animal blood vessel includes one of a straight structure and a bifurcated structure.

[0079] An embodiment of the present application also provides a decellularized artificial lymph drainage tube.

[0080] A decellularized artificial lymph drainage tube is prepared by adopting the above-mentioned preparation method.

[0081] An embodiment of the present application also provides an application of a decellularized artificial lymph drainage tube.

[0082] A decellularized artificial lymphatic drainage tube is used in lymphatic-venous anastomosis, wherein one end of the decellularized artificial lymphatic drainage tube can be anastomosed with an autologous vein or its branch, or with another decellularized artificial lymphatic drainage tube, and the other end of the decellularized artificial lymphatic drainage tube can be anastomosed with an autologous lymphatic vessel, or with another decellularized artificial lymphatic drainage tube or with an opening left in subcutaneous tissue.

[0083] Example 1

[0084] This embodiment provides a decellularized artificial lymph drainage tube.

[0085] The acellular artificial lymph drainage tube of this embodiment is prepared by the following preparation method:

[0086] (1) Obtain a whole piece of subcutaneous connective tissue from the pig's hind limb and separate a pig saphenous artery with a length of approximately 10 cm.

[0087] (2) Using porcine saphenous arteries as raw materials, the obtained porcine saphenous arteries were placed in a 2 wt % NaOH solution and decellularized at room temperature for 4 h.

[0088] (3) Then rinse thoroughly with purified water for 5 minutes.

[0089] (4) The cleaned decellularized porcine saphenous artery was then placed in a 0.05wt% glutaraldehyde solution for cross-linking treatment for 6 hours.

[0090] (5) After the cross-linked decellularized saphenous artery is thoroughly cleaned, a needle with an outer diameter of about 0.5 mm is used to evenly insert holes at intervals of about 0.5 cm on the surface of the cross-linked decellularized abdominal aorta.

[0091] (6) Freeze-drying to obtain a cell-free artificial lymph drainage tube derived from the porcine saphenous artery, see Figure 1 The scanning electron microscopy of the decellularized artificial lymphatic drainage tube derived from the porcine saphenous artery is shown in the attached Figure 2 and attached Figure 3 , Figure 2 The outer surface microstructure of the decellularized artificial lymph drainage tube derived from the porcine saphenous artery of this embodiment; Figure 3 The inner surface microstructure of the decellularized artificial lymph drainage tube derived from the porcine saphenous artery of this embodiment; Figure 4 This is the H&E stained section of the material before decellularization; Figure 5 These are H&E stained sections of decellularized material.

[0092] Example 2

[0093] This embodiment provides a decellularized artificial lymph drainage tube.

[0094] The acellular artificial lymph drainage tube of this embodiment is prepared by the following preparation method:

[0095] (1) Euthanized rats were obtained and the abdominal aorta with a length of about 10 cm was isolated.

[0096] (2) Using the abdominal aorta as raw material, the obtained abdominal aorta was placed in a 0.25wt% pancreatic enzyme solution and decellularized at 33°C for 4 hours.

[0097] (3) Then wash thoroughly with PBS buffer for 10 minutes.

[0098] (4) The cleaned decellularized abdominal aorta was then placed in a 0.01 wt % genipin solution and stirred for cross-linking at room temperature for 2 h.

[0099] (5) After the cross-linked decellularized abdominal aorta is thoroughly washed with pure water, a needle with an outer diameter of about 0.5 mm is used to evenly make holes at intervals of about 0.5 cm on the surface of the cross-linked decellularized abdominal aorta.

[0100] (6) Freeze-drying to obtain a decellularized artificial lymph drainage tube derived from the rat abdominal aorta.

[0101] The performance tests were performed on the acellular artificial lymph drainage tubes in Example 1 and Example 2, and the test results are shown in Table 1.

[0102] Table 1

[0103]

[0104] Among them, the in vitro degradation time is obtained by immersing the material in a 60U / mL collagenase solution and recording the time when the mass decreases by more than 95%.

[0105] The cell viability rate is measured by placing the material in MEM culture medium at a ratio of 0.2 g / mL and immersing it for 72 hours, and then measuring the 24-hour survival rate of mouse fibroblasts in the extract.

[0106] In summary, the above-mentioned decellularized artificial lymph drainage tube can solve the problems of time-consuming and difficult operation of existing lymphatic-venous anastomosis surgery, and can drain the lymph from the swollen part of the patient to the venous system in time after implantation, and can quickly relieve the symptoms of swelling. The decellularized artificial lymph drainage tube of the present application supports the growth, migration, and proliferation of autologous cells, has good biocompatibility, and the degradation time can be adjusted to match the reconstruction time of the autologous lymphatic return system to ensure long-term patency. Specifically, the present application has an artificial lymph drainage tube with suitable degradation performance that simulates the elasticity of blood vessels and is of great significance. The decellularized artificial lymph drainage tube based on animal blood vessels as raw materials not only has excellent biocompatibility, but also can retain the elasticity of the blood vessels themselves; its degradation rate can be adjusted according to the degree of cross-linking process to match the reconstruction rate of the autologous lymphatic return system, thereby ensuring smooth long-term lymph return.

[0107] In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0108] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0109] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for preparing an acellular artificial lymph drainage tube, characterized in that: The steps include: Animal blood vessels are used as raw materials, and the raw materials are subjected to decellularization, cleaning, strengthening, pore making and drying treatments.

2. The method for preparing the acellular artificial lymph drainage tube according to claim 1, characterized in that: The animal blood vessels include one or more blood vessels of pigs, cattle, sheep, rabbits, mice, fish and poultry; And / or, the animal blood vessels include one or a combination of arterial blood vessels and venous blood vessels.

3. The method for preparing the acellular artificial lymph drainage tube according to claim 1, characterized in that: The decellularization treatment includes one or a combination of alkali treatment, acid treatment, surfactant treatment and enzyme treatment.

4. The method for preparing the acellular artificial lymph drainage tube according to claim 3, characterized in that: The alkali treatment comprises the following steps: placing the raw material in an alkali solution with an alkali concentration of 0.5wt%-5wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5h-12h; And / or, the acid treatment comprises the following steps: placing the raw material in an acid solution with an acid concentration of 0.1wt% to 0.5wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5h to 12h; And / or, the surfactant treatment comprises the following steps: placing the raw material in a surfactant solution with a surfactant concentration of 1 wt% to 5 wt%, and performing a decellularization treatment at room temperature or -4°C for 0.5 h to 12 h; And / or, the enzyme treatment comprises the following steps: placing the raw material in a pancreatic enzyme solution with a pancreatic enzyme concentration of 0.25wt% to 1.0wt%, and performing a decellularization treatment at a temperature of 30°C to 35°C for 4h to 24h; And / or, when the decellularization treatment includes a combination of at least two of the alkaline treatment, the acid treatment, the surfactant treatment, and the enzyme treatment, a plurality of washing steps are interspersed between each decellularization treatment step.

5. The method for preparing the acellular artificial lymph drainage tube according to any one of claims 1 to 4, characterized in that: The reinforcing treatment includes one or a combination of a cross-linking treatment and a surface coating treatment.

6. The method for preparing the acellular artificial lymph drainage tube according to claim 5, characterized in that: The cross-linking treatment includes one or a combination of thermal cross-linking and chemical cross-linking.

7. The method for preparing the acellular artificial lymph drainage tube according to claim 6, characterized in that: The thermal cross-linking comprises the following steps: freeze-drying the raw material after the cleaning treatment, and then placing it in a vacuum environment at 60° C. to 120° C. for cross-linking treatment for 1 h to 24 h; And / or, the chemical cross-linking comprises the following steps: placing the raw material after the cleaning treatment in a chemical cross-linking agent solution with a chemical cross-linking agent concentration of 0.01wt% to 5wt%, stirring and cross-linking at room temperature or 4°C for 1h to 72h; And / or, the chemical cross-linking agent includes one or more combinations of glutaraldehyde, formaldehyde, genipin, butanediol glycidyl ether, divinyl sulfone, oxalic acid dihydrazide, carbodiimide and glycidyl methacrylate.

8. The method for preparing the acellular artificial lymph drainage tube according to claim 5, characterized in that: The surface coating treatment comprises the following steps: placing the raw material after the cleaning treatment in a biocompatible polymer solution with a biocompatible polymer concentration of 0.1wt% to 5wt%, soaking for 30s to 10min, then taking out the raw material and removing the solvent of the biocompatible polymer; And / or, the biocompatible polymer includes one or more combinations of chitosan, cellulose, polylactic acid, silk fibroin, collagen, polyvinyl alcohol, polylactic acid-co-glycolic acid, polymethyl methacrylate, polyamide, polyurethane and polycaprolactone.

9. The method for preparing the acellular artificial lymph drainage tube according to any one of claims 1 to 4 and 6 to 8, characterized in that: The length of the animal blood vessel is 0.5 cm to 50 cm; And / or, the outer diameter of the animal blood vessel is 0.1 mm to 6 mm; And / or, the animal blood vessel includes a straight structure and a bifurcated structure.

10. An acellular artificial lymph drainage tube, characterized in that: The method is prepared by any one of claims 1 to 9.

11. Use of the acellular artificial lymphatic drainage tube according to claim 10 in lymphatic-venous anastomosis, wherein: One end of the decellularized artificial lymph drainage tube can be anastomosed with an autologous vein or its branch, or with another decellularized artificial lymph drainage tube, and the other end of the decellularized artificial lymph drainage tube can be anastomosed with an autologous lymphatic vessel, or with another decellularized artificial lymph drainage tube or with an opening left in the subcutaneous tissue.

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