A stapler tissue engineering composite patch and its preparation method and application

Abstract

The invention discloses a method for preparing a stapler tissue engineering composite patch, which comprises removing cells from an extracellular matrix layer by using a chemical reagent until DNA is less than 100 ng / mg and an α-Gal clearance rate is greater than 90%, wherein the extracellular matrix layer comprises a first biogenic material and / or a tissue engineering material; coating the outside of the extracellular matrix layer with a hydrophilic material so that the suture tension is greater than 2N and the tensile strength is greater than>2N; combining and fixing the treated extracellular matrix layer with a base layer so that the tear strength between the two is greater than 2N, removing moisture to a moisture content of 0-30%, and sterilizing to obtain the stapler tissue engineering composite patch, wherein the base layer comprises a synthetic material and / or a second biogenic material.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to a stapler tissue engineering composite patch and a preparation method and application thereof. Background Art

[0002] Staplers are medical devices that replace manual excision and suturing. They are widely used in minimally invasive surgery because of their safety and ease of operation. On the one hand, minimally invasive surgery has the advantages of less surgical trauma, less pain and postoperative complications, and faster patient recovery. It has gradually expanded to various departments. The popularity of minimally invasive surgery will drive the continued growth of the stapler market. On the other hand, with the continuous improvement of China's medical level, doctors and patients have increasingly higher requirements for the accuracy and safety of surgical operations. Staplers can improve surgical efficiency and quality, shorten recovery time, and the development trend of the stapler market continues to improve. The advancement of modern surgical instruments has promoted the advancement of surgical technology. In the field of gastrointestinal surgery, the emergence of staplers has brought digestive tract reconstruction to a new stage, shortened operation time, reduced tissue damage, and shortened hospital stays.

[0003] According to relevant analysis, the global stapler market size will reach US$8.91 billion in 2022, with a compound growth rate of 4.9% from 2017 to 2022, of which the electric laparoscopic stapler market size will reach US$1.66 billion, accounting for 18.7% of the total; the global stapler market size is expected to grow to US$10.43 billion by 2025, with a compound growth rate of 5.4% from 2022 to 2025, of which the market size of electric laparoscopic staplers is expected to grow to US$2.25 billion, accounting for 21.6% of the total; the global stapler market size is expected to further grow to US$12.93 billion by 2030, with a compound growth rate of 4.4% from 2025 to 2030, of which the electric laparoscopic stapler market size is expected to further grow to US$4.33 billion.

[0004] The growth rate of China's stapler market size is higher than the global average. In 2022, the domestic stapler market size will reach 8.63 billion yuan, with a compound growth rate of 12.5% ​​from 2017 to 2022, of which the electric laparoscopic stapler market size will reach 2.82 billion yuan; it is expected that by 2025, the size of China's stapler market will grow to 11.89 billion yuan, with a compound growth rate of 11.3% from 2022 to 2025, of which the proportion of electric laparoscopic staplers is expected to gradually increase, and is expected to grow to 6.51 billion yuan in 2025, accounting for 54.8% of the total; it is expected that by 2030, the size of China's stapler market will further grow to 20.50 billion yuan, with a compound growth rate of 11.5% from 2025 to 2030, of which the size of the electric laparoscopic stapler market is expected to further grow to 12.74 billion yuan, accounting for 62.2% of the total.

[0005] Staplers are used in surgical operations to sew up the anastomosis of tissues, blood vessels, catheters, shunts, or other objects or body parts involved in specific operations. These anastomosis can be channels or internal organs in the body, or they can be punctured tissues and cut tissues or blood vessels formed during the operation to form a bridge or anastomosis. During the driving process of the stapler, tissue movement or slippage caused by the application of pressure may damage the integrity of the staple line. Tissue movement and slippage may increase surgical complications and adverse clinical outcomes, such as surgical bleeding, leakage, and infection, which will not only affect the patient's postoperative healing, but also increase the cost of patients and medical institutions. When the staples of the stapler penetrate the tissue, leakage may occur due to the holes formed by the staples, so that blood, air, gastrointestinal fluid, and other fluids can seep out through the openings formed by the staples, causing trauma and inflammation.

[0006] Stapler patches are used to repair injured or traumatized body tissues and help reconstruct damaged tissues such as tissues, cartilage, skin, and muscles. They not only provide structural support for tissue repair, but also promote "inward" growth of tissues. There are currently two types of patches used clinically, synthetic materials and biomaterials. Synthetic materials are non-absorbable materials of the body. This type of material has poor compliance and can cause discomfort to patients after implantation. Long-term implantation may lead to infection. Biomaterials have good compatibility with the body, but they lack the mechanical properties required for patches and have poor support performance. It is difficult to make biomaterials into precise shapes and thicknesses.

[0007] Currently, adhesions between internal organs and surgical sites are very likely to occur after clinical surgery, which is difficult to avoid even for the most cautious surgeons. Adhesions will form scar tissue at the affected site, causing complications such as intestinal obstruction, female infertility, and chronic debilitating pain. In addition, adhesions increase the medical costs of subsequent surgeries and medications, and will prolong the patient's recovery time and increase postoperative risks. Summary of the invention

[0008] The main purpose of the present invention is to provide a stapler tissue engineering composite patch and its preparation method and application, aiming to solve the above problems.

[0009] To achieve the above object, the present invention provides a method for preparing a stapler tissue engineering composite patch, the method for preparing the stapler tissue engineering composite patch comprising:

[0010] Using chemical reagents to remove cells from the extracellular matrix layer until DNA is less than 100 ng / mg and α-Gal clearance rate is greater than 90%, wherein the extracellular matrix layer includes a first biogenic material and / or a tissue engineering material;

[0011] The hydrophilic material is coated on the outside of the extracellular matrix layer so that the suture tension is greater than 2N and the tensile strength is greater than 2N;

[0012] The treated extracellular matrix layer is combined and fixed with the base layer to achieve a tear strength greater than 2N, moisture is removed to a moisture content of 0-30%, and after sterilization, a stapler tissue engineering composite patch is obtained, wherein the base layer includes a synthetic material and / or a second biogenic material.

[0013] Preferably, in the method for preparing the stapler tissue engineering composite patch, the first biogenic material / the second biogenic material is a mammalian material;

[0014] The tissue engineering material is prepared by planting seed cells in a degradable polymer material for cultivation.

[0015] Preferably, in the method for preparing the stapler tissue engineering composite patch, the first biogenic material / the second biogenic material is one or more of liver, pericardium, connective tissue, peritoneum, dura mater, and dermis of a mammal;

[0016] The degradable polymer material used in the tissue engineering material is one or more of polylactic acid, polyvinyl alcohol, polycarbonate, polyglycolide, polyglycolic acid, and polylactide.

[0017] Preferably, in the method for preparing the stapler tissue engineering composite patch, the extracellular matrix layer includes one or more of collagen, elastin, fibronectin, fibrin, laminin, extracellular matrix-associated growth proteins, glycosaminoglycans, matrix materials, reticular fibers and thrombospondin.

[0018] Preferably, in the method for preparing the stapler tissue engineering composite patch, in the step of removing cells from the extracellular matrix layer using a chemical reagent until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90%, the chemical reagent is a detergent, and the detergent includes a non-ionic detergent and a zwitterionic detergent;

[0019] Accordingly, the method of removing cells from the extracellular matrix layer using chemical reagents until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90% comprises:

[0020] The cells in the extracellular matrix layer are removed by detergent until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90%. The treatment temperature for removing cells by detergent is 25-37°C, the treatment time is 2-72 hours, and the detergent-containing solution is replaced every 2-12 hours.

[0021] Preferably, in the method for preparing the stapler tissue engineering composite patch, in the step of coating the outside of the extracellular matrix layer with a hydrophilic material so that the suture tension is greater than 2N and the tensile strength is greater than 2N, the hydrophilic material is one or more of hyaluronic acid, carrageenan, gelatin, oxidized regenerated cellulose, and collagen;

[0022] and / or,

[0023] The concentration of the hydrophilic material is 0.01-2%.

[0024] Preferably, in the preparation method of the stapler tissue engineering composite patch, in the step of combining and fixing the treated extracellular matrix layer and the base layer so that the tear strength between the two is greater than 2N, removing moisture to a moisture content of 0-30%, and sterilizing to obtain the stapler tissue engineering composite patch, the base layer is a synthetic material or a second biogenic material, the synthetic material is a mixture of degradable polymer fibers and non-degradable polymer fibers, the content of the degradable polymer fibers is greater than the content of the non-degradable polymer fibers, and the density of the synthetic material is 20 mg / cm 3 ~500mg / cm 3 ;

[0025] Wherein, the degradable polymer is one or more of polylactic acid, polyphosphate, polyamino acid, polyvinyl alcohol, polycarbonate, polylactic acid, polyglycolic acid, polyamide, pectin, fibrin, and chitosan;

[0026] The non-degradable polymer fiber is one or more of polypropylene, polyethylene, polyester, nylon, polyacrylate, polymethacrylate, polyurethane, polystyrene, polyvinyl chloride, vinyl fluoride, ePTFE and cellulose acetate;

[0027] The second biogenic material is one or more of mammalian dermis, connective tissue, pericardium, and peritoneum.

[0028] Preferably, in the method for preparing the stapler tissue engineering composite patch, the synthetic material is a filamentous polymer material, the filament diameter of the filamentous polymer material is 10 to 100 deniers, and the filamentous polymer material is prepared by weaving, non-woven to prepare a sheet or mesh structure, solution casting, electrospinning, 3D printing, injection molding, blow molding, extrusion; and / or,

[0029] The two polymers in the composite material have different melting points.

[0030] Preferably, in the method for preparing the stapler tissue engineering composite patch, in the step of combining and fixing the treated extracellular matrix layer and the basal layer so that the tear strength between the two is greater than 2N, removing moisture to a moisture content of 0-30%, and sterilizing to obtain the stapler tissue engineering composite patch, the extracellular matrix layer and the basal layer are laid in the following manner: one or more layers of the treated extracellular matrix layer are laid on top of one or more layers of the basal layer; and / or,

[0031] The combination of the extracellular matrix layer and the basal layer is fixed by sutures or coating with hydrogel; and / or,

[0032] The moisture is removed by drying or freeze-drying. When drying is used to remove moisture, the drying temperature is 25 to 37° C. and the drying time is 1 to 48 hours. When freeze-drying is used to remove moisture, the freeze-drying temperature is -20° C. to -80° C. and the freeze-drying time is 1 to 96 hours.

[0033] In order to achieve the above-mentioned purpose, the present invention further provides a stapler tissue engineering composite patch, which is prepared by the above-mentioned stapler tissue engineering composite patch preparation method.

[0034] In order to achieve the above object, the present invention also provides an application of the above stapler tissue engineering composite patch, wherein the stapler tissue engineering composite patch is used for reinforcement during gastrectomy, gastric banding, small intestine, colon, and colorectal surgery; and / or,

[0035] The stapler tissue engineering composite patch is used for reinforcement in lung resection and other lung and bronchial incisions; and / or,

[0036] The stapler tissue engineering composite patch is used in gynecology, neurosurgery, arthroscopy, orthopedics, plastic surgery, reconstruction, muscle and tendon surgery.

[0037] The present invention has at least the following beneficial effects:

[0038] The preparation method of the stapler tissue engineering composite patch provided by the present invention comprises the following steps: removing cells from the extracellular matrix layer using chemical reagents until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90%, wherein the extracellular matrix layer comprises a first biogenic material and / or a tissue engineering material; coating the outside of the extracellular matrix layer with a hydrophilic material so that the suture tension is greater than 2N and the tensile strength is greater than>2N; combining and fixing the treated extracellular matrix layer with a base layer so that the tear strength between the two is greater than 2N, removing moisture to a moisture content of 0-30%, and sterilizing to obtain the stapler tissue engineering composite patch, wherein the base layer comprises a synthetic material and / or a second biogenic material. The composite patch provided in this way can support tissue reconstruction, resist infection, and have no immunogenicity, and has both strength and toughness, excellent mechanical properties, and completely degrades within a certain period of time after being implanted in the body, thereby reducing scarring and fibrosis of the affected area during wound healing.

[0039] Furthermore, the present invention uses degradable polymer materials / biomaterials as the base layer and biogenic / tissue engineering materials as the matrix layer. It has good biocompatibility and reduces immunogenicity after removing cells. While providing mechanical properties and strengthening tissues, the extracellular matrix provides tissue strengthening, reduces inflammation, and promotes cell growth, effectively inhibiting fibrosis, scar formation and surgical adhesions. Its thickness can also be applied to different surgical sites.

[0040] Furthermore, in the solution provided by the present invention, the extracellular matrix material is not chemically cross-linked and can be completely absorbed by the body within a certain period of time after being implanted in the body. It is not a permanent implant in the traditional sense. The tissue engineering material is not of animal origin, eliminating the risk of viral infection. The tissue engineering material is cultured on a large scale in vitro and is completely renewable, and is not affected by animal species and individual differences. The size and thickness of the material can be customized according to needs and can be used on demand, thereby expanding the scope of clinical use of patches. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0042] Figure 1 The present invention is a schematic flow chart of an embodiment of a method for preparing a stapler tissue engineering composite patch provided by the present invention.

[0043] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0044] To make the purpose, technical scheme and advantages of the embodiment of the present invention clearer, the technical scheme in the embodiment of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiment, it is carried out according to the normal conditions or the conditions recommended by the manufacturer. If the reagents or instruments used do not specify the manufacturer, they are all conventional products that can be purchased commercially. In addition, the meaning of "and / or" appearing in the full text includes three parallel schemes, taking "A and / or B" as an example, including scheme A, or scheme B, or schemes that A and B meet at the same time. In addition, the technical schemes between the various embodiments can be combined with each other, but must be based on the ability of ordinary technicians in the field to achieve. When the combination of technical solutions is contradictory or cannot be achieved, it should be considered that the combination of such technical solutions does not exist, and is not within the scope of protection required by the present invention. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work belong to the scope of protection of the present invention.

[0045] Figure 1 The figure shows a flow chart of an embodiment of a method for preparing a stapler tissue engineering composite patch provided by the present invention. Figure 1 In step S100, cells are removed from the extracellular matrix layer using chemical reagents until DNA is less than 100 ng / mg and α-Gal clearance is greater than 90%, wherein the extracellular matrix layer includes a first biogenic material and / or a tissue engineering material.

[0046] The concentration of DNA may be 90 ng / mg, 80 ng / mg, 70 ng / mg, etc. The α-Gal clearance rate may be 92%, 94%, 96% or 100%.

[0047] The first biogenic material may be a mammalian material. Specifically, the first biogenic material is one or more of the liver, pericardium, connective tissue, peritoneum, dura mater, and dermis of a mammal. The mammalian material may be a mammalian material derived from cattle, pigs, dogs, horses, sheep, etc., and is not specifically limited here.

[0048] The tissue engineering material is prepared by planting seed cells in a degradable polymer material and then culturing the same. Specifically, the degradable polymer material used can be, but is limited to, one or more of polylactic acid, polyvinyl alcohol, polycarbonate, polyglycolide, polyglycolic acid, and polylactide.

[0049] More specifically, the extracellular matrix layer includes one or more of collagen, elastin, fibronectin, fibrin, laminin, extracellular matrix-related growth proteins (such as growth factors and cytokines), glycosaminoglycans, matrix substances, reticular fibers, and thrombospondin.

[0050] Wherein, the chemical reagent is a detergent, and the detergent includes a non-ionic detergent and a zwitterionic detergent; accordingly, step S100 includes:

[0051] The cells in the extracellular matrix layer are removed by detergent until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90%. The treatment temperature for removing cells by detergent is 25-37°C, the treatment time is 2-72 hours, and the detergent-containing solution is replaced every 2-12 hours.

[0052] The processing temperature may also be 28°C, 30°C, or 35°C.

[0053] The processing time can also be 10h, 20h, 30h, 40h, or 50h.

[0054] The time interval for replacing the detergent-containing solution may also be 5h, 8h, or 10h.

[0055] Step S200, coating the outside of the extracellular matrix layer with a hydrophilic material, so that the suture tension is greater than 2N and the tensile strength is greater than 2N. The hydrophilic material may be, but is not limited to, one or more of hyaluronic acid, carrageenan, gelatin, oxidized regenerated cellulose, and collagen. Preferably, the concentration of the hydrophilic material is 0.01-2%. The concentration of the hydrophilic material may also be 0.1%, 0.5%, 1%, or 1.5%.

[0056] Step S300, the treated extracellular matrix layer is combined and fixed with the base layer so that the tear strength between the two is greater than 2N, moisture is removed to a moisture content of 0-30%, and after sterilization, a stapler tissue engineering composite patch is obtained, wherein the base layer includes a synthetic material and / or a second biogenic material.

[0057] The base layer is a synthetic material or a second biogenic material, the synthetic material is a mixture of degradable polymer fibers and non-degradable polymer fibers, the content of the degradable polymer fibers is greater than the content of the non-degradable polymer fibers, and the density of the synthetic material is 20 mg / cm 3 ~500mg / cm 3 .

[0058] The density of the synthetic material can also be 100mg / cm 3 、200mg / cm 3 、300mg / cm 3 , or 400mg / cm 3 .

[0059] It should be noted that the selection of the second biogenic material can refer to the above-mentioned first biogenic material, that is, the second biogenic material can also be a mammalian material. Specifically, the second biogenic material is one or more of the liver, pericardium, connective tissue, peritoneum, dura mater, and dermis of a mammal. The mammalian material can be a mammalian material derived from cattle, pigs, dogs, horses, sheep, etc., and is not specifically limited here.

[0060] When making a specific selection, the first biogenic material and the second biogenic material may be made of the same material or different materials, which may be set according to needs.

[0061] Wherein, the degradable polymer is one or more of polylactic acid, polyphosphate, polyamino acid, polyvinyl alcohol, polycarbonate, polylactic acid, polyglycolic acid, polyamide, pectin, fibrin, and chitosan. The non-degradable polymer fiber is one or more of polypropylene, polyethylene, polyester, nylon, polyacrylate, polymethacrylate, polyurethane, polystyrene, polyvinyl chloride, vinyl fluoride, ePTFE, and cellulose acetate. The second biogenic material is one or more of mammalian dermis, connective tissue, pericardium, and peritoneum.

[0062] The synthetic material is a filamentous polymer material, the filament diameter of the filamentous polymer material is 10 to 100 deniers, and the filamentous polymer material is prepared by weaving, non-woven to prepare a sheet or mesh structure, solution casting, electrostatic spinning, 3D printing, injection molding, blow molding, and extrusion. The two polymers in the synthetic material have different melting points.

[0063] The laying method of the extracellular matrix layer and the base layer can be, but is not limited to, laying one or more layers of treated extracellular matrix layer on top of one or more layers of base layer, or the combination of the extracellular matrix layer and the base layer can be fixed by sutures or coated with hydrogel.

[0064] In addition, in step S300, the water is removed by drying or freeze drying. When drying is used to remove water, the drying temperature is 25-37° C. and the drying time is 1-48 hours. The drying temperature can also be 28° C., 30° C., 33° C., or 35° C. The drying time can also be 10 hours, 20 hours, 30 hours, or 40 hours.

[0065] When freeze drying is used to remove moisture, the freeze drying temperature is -20°C to -80°C, and the freeze drying time is 1 to 96 hours. The freeze drying temperature can also be -30°C, -40°C, -50°C or -60°C. The freeze drying time can also be 10 hours, 30 hours, 50 hours, 70 hours, or 90 hours.

[0066] The present invention also provides a stapler tissue engineering composite patch, which is prepared by using the above-mentioned stapler tissue engineering composite patch preparation method.

[0067] The present invention also provides an application of the stapler tissue engineering composite patch, which is used for reinforcement during gastrectomy, gastric banding, small intestine, colon, and colorectal surgery. The stapler tissue engineering composite patch can also be used for reinforcement during lung resection and other lung and bronchial incisions. The stapler tissue engineering composite patch can also be used in gynecology, neurosurgery, arthroscopy, orthopedics, plastic surgery, reconstruction, muscle, and tendon surgery.

[0068] In order to verify the beneficial effects of the solution provided by the present invention, examples and comparative examples are used for verification.

[0069] Example 1

[0070] (1) Obtain the pericardium of a healthy, quarantined cow weighing 900 kg from a slaughterhouse. Use medical scissors to carefully remove the fat and connective tissue outside the pericardium to obtain the extracellular matrix layer;

[0071] (2) The obtained extracellular matrix layer was treated with 500 mmol / L polyoxyethyleneamide and 800 mmol / L deoxycholine sulfobetaine at 37°C for 48 h, with the liquid replaced every 6 h. The DNA content was 57 ng / mg and the α-Gal clearance rate was 96.2%;

[0072] (3) After the bovine pericardium was soaked in 0.05% hyaluronic acid with a molecular weight of 500,000 for 4 hours, the suture tension was 3.6±0.2N and the tensile strength was 2.83±0.31N;

[0073] (4) A woven polypropylene polymer layer was used as the base layer with a thickness of 0.2 mm. One layer of polypropylene and one layer of bovine pericardium were superimposed and sutured using 4-0 sutures at the edges of the sutures. The tear strength of polypropylene and bovine pericardium was 4.33±1.27N. The composite patch was freeze-dried at -60°C for 48 hours with a moisture content of 0.5%. After double-layer packaging and irradiation sterilization, a stapler tissue engineering composite patch was obtained.

[0074] Example 2

[0075] (1) Obtain the small intestine of a healthy, quarantined pig weighing 700 kg from a slaughterhouse. Use medical scissors to carefully remove the fat and connective tissue outside the small intestine, separate the small intestinal submucosa, and obtain the extracellular matrix layer;

[0076] (2) The obtained extracellular matrix layer was treated with 400 mmol / L alkyl polyglycosides and 700 mmol / L lauryl sulfobetaine at 25°C for 48 h, with the liquid replaced every 4 h. The DNA content was 65 ng / mg and the α-Gal clearance rate was 92.8%;

[0077] (3) The decellularized small intestinal submucosa was treated with 20% carboxyl content and 0.5 mg / ml oxidized carboxycellulose for 6 h, and the suture tension was 3.78 ± 0.21 N;

[0078] (4) 200°C injection molded polyethylene with a thickness of 0.3 mm was used as the base layer, and two layers of small intestinal submucosal layer were used as the extracellular matrix layer. The edges were sutured with 4-0 sutures, with a tear strength of 3.21±0.36N. After freeze-drying at -60°C for 72h, the moisture content was 0.76%, and the composite patch was sterilized with propylene oxide after double-layer packaging to form a stapler tissue engineering composite patch.

[0079] Comparative Example 1

[0080] (1) Obtain the peritoneum of healthy sheep weighing 200 kg from a slaughterhouse. Carefully remove the fat and connective tissue outside the peritoneum using medical scissors;

[0081] (2) The peritoneum was treated with 300mmol / L monoglyceride and 900mmol / L myristoyl sulfobetaine for 36 hours, with the fluid replaced every 12 hours; the DNA content was 125ng / mg, and the α-Gal clearance rate was 72.8%

[0082] (3) The decellularized peritoneum was treated with 10% gelatin solution for 3 h, with a suture tension of 1.51 ± 0.32 N and a leakage of 0.5 mg / min at 120 mmHg;

[0083] (4) A 300 μm thick PGA substrate layer was prepared from a 6% PGA electrospinning solution. After the PGA layer was sutured to the peritoneum using a 4-0 suture thread, the tear strength was 1.51±0.21 N. After drying at 25°C for 12 h, the moisture content was 11.2%.

[0084] Table 1 Comparison of effects of embodiments and comparative examples

[0085]

[0086] It can be seen from Table 1 and the above embodiments and comparative examples that the solution provided by the present invention has high conformity (no leakage), is anti-infective, non-immunogenic, has both strength and toughness, and has excellent mechanical properties. It is completely degraded within a certain period of time after implantation in the body, and reduces scarring and fibrosis of the affected area during wound healing.

[0087] The above are only preferred embodiments of the present invention, and do not limit the patent scope of the present invention. For those skilled in the art, the present invention may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the patent protection scope of the present invention.

Claims

1. A method for preparing a stapler tissue engineering composite patch, characterized in that: include: Using chemical reagents to remove cells from the extracellular matrix layer until DNA is less than 100 ng / mg and α-Gal clearance rate is greater than 90%, wherein the extracellular matrix layer includes a first biogenic material and / or a tissue engineering material; The hydrophilic material is coated on the outside of the extracellular matrix layer, so that the suture tension is greater than 2N and the tensile strength is greater than 2N; The treated extracellular matrix layer is combined and fixed with the base layer to achieve a tear strength greater than 2N, moisture is removed to a moisture content of 0-30%, and after sterilization, a stapler tissue engineering composite patch is obtained, wherein the base layer includes a synthetic material and / or a second biogenic material.

2. The method for preparing the stapler tissue engineering composite patch according to claim 1, characterized in that: The first biogenic material / the second biogenic material is a mammalian material; The tissue engineering material is prepared by planting seed cells in a degradable polymer material for cultivation.

3. The method for preparing the stapler tissue engineering composite patch according to claim 2, characterized in that: The first biogenic material / the second biogenic material is one or more of liver, pericardium, connective tissue, peritoneum, dura mater, and dermis of a mammal; The degradable polymer material used in the tissue engineering material is one or more of polylactic acid, polyvinyl alcohol, polycarbonate, polyglycolide, polyglycolic acid, and polylactide.

4. The method for preparing the stapler tissue engineering composite patch according to claim 1, characterized in that: The extracellular matrix layer includes one or more of collagen, elastin, fibronectin, fibrin, laminin, extracellular matrix-related growth protein, glycosaminoglycan, matrix material, reticular fiber and thrombospondin.

5. The method for preparing the stapler tissue engineering composite patch according to claim 1, characterized in that: In the step of removing cells from the extracellular matrix layer using chemical reagents until DNA is less than 100 ng / mg and α-Gal clearance is greater than 90%, the chemical reagent is a detergent, and the detergent includes a non-ionic detergent and a zwitterionic detergent; Accordingly, the method of removing cells from the extracellular matrix layer using chemical reagents until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90% comprises: The cells in the extracellular matrix layer are removed by detergent until DNA is less than 100 ng / mg and the α-Gal clearance rate is greater than 90%. The treatment temperature for removing cells by detergent is 25-37°C, the treatment time is 2-72 hours, and the detergent-containing solution is replaced every 2-12 hours.

6. The method for preparing the stapler tissue engineering composite patch according to claim 1, characterized in that: In the step of coating the outside of the extracellular matrix layer with a hydrophilic material so that the suture tension is greater than 2N and the tensile strength is greater than 2N, the hydrophilic material is one or more of hyaluronic acid, carrageenan, gelatin, oxidized regenerated cellulose, and collagen; and / or, The concentration of the hydrophilic material is 0.01-2%.

7. The method for preparing the stapler tissue engineering composite patch according to claim 1, characterized in that: In the step of combining and fixing the treated extracellular matrix layer and the base layer so that the tear strength between the two is greater than 2N, removing moisture to a moisture content of 0-30%, and sterilizing to obtain a stapler tissue engineering composite patch, the base layer is a synthetic material or a second biogenic material, the synthetic material is a mixture of degradable polymer fibers and non-degradable polymer fibers, the content of the degradable polymer fibers is greater than the content of the non-degradable polymer fibers, and the density of the synthetic material is 20 mg / cm 3 ~500mg / cm 3 ; Wherein, the degradable polymer is one or more of polylactic acid, polyphosphate, polyamino acid, polyvinyl alcohol, polycarbonate, polylactic acid, polyglycolic acid, polyamide, pectin, fibrin, and chitosan; The non-degradable polymer fiber is one or more of polypropylene, polyethylene, polyester, nylon, polyacrylate, polymethacrylate, polyurethane, polystyrene, polyvinyl chloride, vinyl fluoride, ePTFE and cellulose acetate; The second biogenic material is one or more of mammalian dermis, connective tissue, pericardium, and peritoneum.

8. The method for preparing the stapler tissue engineering composite patch according to claim 7, characterized in that: The synthetic material is a filamentous polymer material, the filament diameter of the filamentous polymer material is 10 to 100 deniers, and the filamentous polymer material is prepared by weaving, non-woven to prepare a sheet or mesh structure, solution casting, electrostatic spinning, 3D printing, injection molding, blow molding, extrusion; and / or, The two polymers in the composite material have different melting points.

9. The method for preparing the stapler tissue engineering composite patch according to claim 7, characterized in that: In the step of combining and fixing the treated extracellular matrix layer and the basal layer so that the tear strength between the two is greater than 2N, removing moisture to a moisture content of 0-30%, and sterilizing to obtain a stapler tissue engineering composite patch, the extracellular matrix layer and the basal layer are laid in the following manner: one or more layers of the treated extracellular matrix layer are laid on top of one or more layers of the basal layer; and / or, The combination of the extracellular matrix layer and the basal layer is fixed by sutures or coating with hydrogel; and / or, The moisture is removed by drying or freeze-drying. When drying is used to remove moisture, the drying temperature is 25 to 37° C. and the drying time is 1 to 48 hours. When freeze-drying is used to remove moisture, the freeze-drying temperature is -20° C. to -80° C. and the freeze-drying time is 1 to 96 hours.

10. A stapler tissue engineering composite patch, characterized in that: The stapler tissue engineering composite patch is prepared by the preparation method of the stapler tissue engineering composite patch according to any one of claims 1 to 9.

11. An application of the stapler tissue engineering composite patch as claimed in claim 10, characterized in that: The stapler tissue engineering composite patch is used for reinforcement during gastrectomy, gastric banding, small intestine, colon, and colorectal surgery; and / or, The stapler tissue engineering composite patch is used for reinforcement in lung resection and other lung and bronchial incisions; and / or, The stapler tissue engineering composite patch is used in gynecology, neurosurgery, arthroscopy, orthopedics, plastic surgery, reconstruction, muscle and tendon surgery.

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