Albumin medical tissue adhesive and preparation method and use thereof

Abstract

The present invention discloses a medical tissue adhesive containing recombinant human albumin and a preparation method thereof, belonging to the field of biomedical materials. The recombinant human albumin medical tissue adhesive of the present invention is cross-linked via non-covalent interactions between recombinant human albumin and polyethylene glycol, resulting in strong interfacial adhesion and internal cohesion. The adhesive has a simple preparation process, good biocompatibility and biodegradability, and can provide non-invasive adhesion and immediate sealing, making it suitable for skin cosmetic repair and postoperative wound healing.

Description

Technical Field

[0001] The present invention relates to the field of novel biomaterials, in particular to an albumin medical tissue adhesive and a preparation method and application thereof. Background Art

[0002] Medical tissue adhesives have gained widespread attention in the medical field due to their significant advantages such as ease of operation, non-invasive bonding, instant sealing and hemostasis, and effective shortening of surgical time. They have gradually become an effective auxiliary or alternative to traditional surgical sutures. Currently, a variety of commercial medical tissue adhesives based on various materials have been approved for the repair and regeneration of soft tissues such as the heart, blood vessels, and lungs. However, most of the existing commercially available medical tissue adhesives still face challenges such as insufficient bonding strength, single function, expensive raw materials, poor biocompatibility, and safety risks. The clinical demand for the development of new, efficient, multifunctional soft tissue adhesives is extremely urgent.

[0003] Albumin is one of the most important proteins in animal plasma and is able to maintain the body's nutrition and osmotic pressure. Currently, a variety of albumin-based medical tissue adhesives have been developed. BioGlue (CryoLife Inc.) is an FDA-approved albumin adhesive that can be used to repair vascular injuries during open surgery on large blood vessels (such as the aorta, femoral artery, and carotid artery). Bioglue is made from albumin and glutaraldehyde, and in practical applications, the adhesive's bonding strength can be optimized by varying the ratio and concentration of the two components [Journal of Cardiac Surgery, 2003, 18(6):500-503.]. However, because albumin is typically extracted from animals, it can cause a range of allergic reactions and some blood-borne diseases, posing potential safety risks and limiting its widespread application.

[0004] PEG-based medical tissue adhesives are mainly composed of chemically modified linear or branched PEG molecules [Advanced Materials, 2019, 31(49): e201905761; Biomacromolecules, 2018, 19(3): 980-988]. Coseal glue (Cohesion Technologies, Inc.) is an FDA-approved PEG adhesive for sealing vascular wounds. It is composed of two chemically modified four-arm PEGs, one of which is modified with glutaric acid-succinimidyl ester and the other with thiol. After the two are mixed with a double-barreled syringe, a chemical reaction occurs between the thiol and the carbonyl groups of the succinimidyl ester to form a cross-linked network [Journal of Biomedical Materials Research, 2001, 58(5): 545-555]. Duraseal glue (Covidien Inc.) is another FDA-approved PEG-based adhesive, composed of PEG ester and trilysine amine solution, and is commonly used to seal incisions and anastomoses after neurosurgery to prevent cerebrospinal fluid leakage [Journal of Cardiac Surgery, 2003, 18(6): 504-506]. PEG-based medical tissue adhesives have the advantages of fast curing rate, strong adhesion to biological surfaces, good biocompatibility, and almost no inflammatory response. However, their high equilibrium swelling rate (greater than 400%) may cause severe compression of surrounding nerves and blood vessels; at the same time, excessive swelling of the hydrogel will also lose some mechanical strength and may not be able to maintain its structural integrity in the later stages of wound repair [Chemical Society Reviews, 2015, 44(7): 1820-1835].

[0005] In summary, all types of medical tissue adhesives, including commercial adhesives that have been approved by the FDA, have more or less some defects, such as insufficient bonding strength, poor viscoelasticity of the bonding site, single function, potential safety hazards, etc. The challenges currently faced in clinical applications and scientific research mainly include: ① Most commercial adhesives have a single function and cannot provide more auxiliary treatment effects while achieving wound adhesion, which limits their wide application in clinical medicine; ② Their own mechanical strength is insufficient, and it is urgent to improve the elasticity, toughness and deformation resistance of the adhesive body while ensuring high-strength interfacial adhesion to achieve strong adhesion in a highly dynamic physiological environment; ③ The abundant blood and tissue fluid in the complex physiological environment hinder the rapid, strong and long-lasting adhesion of the adhesive to the biological tissue, and it is extremely difficult to bond the moist tissue surface. Therefore, it is urgent to develop medical tissue adhesives with high mechanical strength, good bonding properties and high biosafety in clinical practice. Summary of the Invention

[0006] Purpose of the invention: To provide a more effective albumin medical tissue adhesive and its preparation method and use. For specific purposes, see the multiple substantial technical effects in the specific implementation section.

[0007] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0008] An albumin medical tissue adhesive, characterized in that the adhesive comprises recombinant human albumin, polyethylene glycol or a polyethylene glycol derivative.

[0009] A further technical solution of the present invention is that the recombinant human albumin and polyethylene glycol are cross-linked by non-covalent interaction; the mass ratio of the recombinant human albumin to the polyethylene glycol is (35-25):(25-35); and the non-covalent interaction is at least one of hydrogen bonding, hydrophobic interaction and van der Waals force.

[0010] A further technical solution of the present invention is that the recombinant human albumin is recombinant human blood albumin, recombinant human serum albumin or a mixture of the two obtained by genetic engineering technology.

[0011] A further technical solution of the present invention is that the genetic engineering technology refers to the recombinant human albumin, recombinant human serum albumin or a mixture of the two obtained by using Pichia pastoris, transgenic animals and plants heterologous expression system.

[0012] A further technical solution of the present invention is that the ratio of the number of free thiol groups contained in the recombinant human albumin is not less than 80% of the total number of free radicals.

[0013] A further technical solution of the present invention is that the molecular weight of the polyethylene glycol or polyethylene glycol derivative is 200-4000; the polyethylene glycol is one or a combination of PEG200, PEG400, PEG600, PEG800, PEG1000, PEG2000, and PEG4000.

[0014] The method for preparing the albumin medical tissue adhesive as described in any of the above items is characterized in that it comprises the following steps: (1) dissolving the recombinant human albumin according to the composition as described in any of the above items to obtain a recombinant human albumin solution; (2) adding the polyethylene glycol to the recombinant human albumin solution in step (1); and (3) cooling to obtain the albumin medical tissue adhesive.

[0015] A further technical solution of the present invention is that in step (1), the dissolution temperature of the recombinant human albumin is less than 60°C; and in step (3), the temperature for dissolving the polyethylene glycol is less than 60°C.

[0016] Use of recombinant human albumin, polyethylene glycol or polyethylene glycol derivatives or the albumin medical tissue adhesive as described in any one of the above items in the preparation of medical repair materials or cosmetic repair materials.

[0017] The preparation method of albumin medical tissue adhesive is characterized by adopting any one of the following methods:

[0018] The first one:

[0019] (1) Weigh 35 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 35%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution;

[0020] (2) Add 25 g of polyethylene glycol (PEG) to the 35% rHSA aqueous solution in portions under heating and stirring at a temperature below 60°C until the PEG is completely dissolved;

[0021] (3) cooling to room temperature to obtain rHSA-PEG adhesive;

[0022] The second type:

[0023] (1) Weigh 33 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 33%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution;

[0024] (2) Add 27 g of polyethylene glycol (PEG) to the 33% rHSA aqueous solution in portions under heating and stirring at a temperature below 60°C until the PEG is completely dissolved;

[0025] (3) cooling to room temperature to obtain rHSA-PEG adhesive;

[0026] The third type:

[0027] (1) Weigh 30 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare a rHSA aqueous solution with a mass volume concentration of 30%. The mixed solution was slowly stirred at room temperature to form a uniform solution;

[0028] (2) Add 30 g of polyethylene glycol (PEG) to the 30% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved;

[0029] (3) cooling to room temperature to obtain rHSA-PEG adhesive;

[0030] The fourth type:

[0031] (1) Weigh 27 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 27%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution;

[0032] (2) Add 33 g of polyethylene glycol (PEG) to the 27% rHSA aqueous solution in portions under heating and stirring at a temperature below 60°C until the PEG is completely dissolved;

[0033] (3) cooling to room temperature to obtain rHSA-PEG adhesive;

[0034] Fifth type:

[0035] (1) Weigh 25 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 25%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution;

[0036] (2) Add 35 g of polyethylene glycol (PEG) to the 25% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved;

[0037] (3) Cooling to room temperature to obtain rHSA-PEG adhesive.

[0038] The present invention, employing the above technical solution, has the following advantages over existing technologies: 1. The albumin medical tissue adhesive of the present invention is cross-linked through physical non-covalent interactions, resulting in strong interfacial bonding and intramolecular cohesion. 2. The albumin medical tissue adhesive of the present invention exhibits excellent biocompatibility and biodegradability. 3. The preparation process of the albumin medical tissue adhesive of the present invention is simple, enabling scalable production. 4. The albumin medical tissue adhesive of the present invention can be used for cosmetic skin repair and postoperative wound healing. DETAILED DESCRIPTION

[0039] In order to make the invention purpose, technical scheme and technical effect of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described in this specification are only for the purpose of explaining the present invention and are not intended to limit the present invention. The present invention is further described in detail below in conjunction with specific examples.

[0040] The present invention aims to provide a medical tissue adhesive with high mechanical strength, excellent bonding properties, and high biosafety, and a method for its preparation. This adhesive uses albumin as the primary raw material, addressing the problem of excessive swelling in polyethylene glycol-based adhesives, which can lead to reduced mechanical strength during use. The adhesive utilizes recombinant human albumin, produced using genetic engineering techniques. Compared to the currently widely used blood-derived albumin, this adhesive contains a higher proportion of free thiol groups, which can enhance the degree of crosslinking with human tissue and the adhesive itself during use. Furthermore, the use of recombinant human albumin can mitigate potential safety hazards associated with blood-borne diseases. The adhesive preparation process is simple to operate and its quality is controllable.

[0041] A method for preparing the above-mentioned albumin medical tissue adhesive comprises the following steps:

[0042] (1) dissolving the recombinant human albumin according to the composition of the albumin medical tissue adhesive to obtain a recombinant human albumin solution;

[0043] (2) adding the polyethylene glycol to the recombinant human albumin solution described in step (1);

[0044] (3) Cooling at room temperature to obtain albumin medical tissue adhesive.

[0045] The polyethylene glycol is added in steps (2) above; preferably, the polyethylene glycol is added in steps (2) above, and is added in 5-15 steps; more preferably, the polyethylene glycol is added in steps (2) above, and is added in 8-10 steps.

[0046] Preferably, the temperature for dissolving polyethylene glycol in step (3) is less than 60°C.

[0047] An application of the above-mentioned albumin medical tissue adhesive in the preparation of medical repair materials.

[0048] The medical repair materials are skin beauty repair materials and postoperative wound healing materials.

[0049] Example 1:

[0050] This example prepares an albumin medical tissue adhesive, and the specific process is as follows:

[0051] (1) Weigh 35 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 35%. The mixed solution was slowly stirred at room temperature to form a uniform solution.

[0052] (2) Add 25 g of polyethylene glycol (PEG) to the above 35% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved.

[0053] (3) Cooling to room temperature to obtain rHSA-PEG adhesive.

[0054] Example 2:

[0055] This example prepares an albumin medical tissue adhesive, and the specific process is as follows:

[0056] (1) Weigh 33 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 33%. The mixed solution was slowly stirred at room temperature to form a uniform solution.

[0057] (2) Add 27 g of polyethylene glycol (PEG) to the above 33% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved.

[0058] (3) Cooling to room temperature to obtain rHSA-PEG adhesive.

[0059] Example 3:

[0060] This example prepares an albumin medical tissue adhesive, and the specific process is as follows:

[0061] (1) Weigh 30 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 30%. The mixed solution is slowly stirred at room temperature to form a uniform solution.

[0062] (2) Add 30 g of polyethylene glycol (PEG) to the above 30% rHSA aqueous solution in portions under heating and stirring at a temperature lower than 60° C. until the PEG is completely dissolved.

[0063] (3) Cooling to room temperature to obtain rHSA-PEG adhesive.

[0064] Example 4:

[0065] This example prepares an albumin medical tissue adhesive, and the specific process is as follows:

[0066] (1) Weigh 27 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 27%. The mixed solution is slowly stirred at room temperature to form a uniform solution.

[0067] (2) Add 33 g of polyethylene glycol (PEG) to the above 27% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved.

[0068] (3) Cooling to room temperature to obtain rHSA-PEG adhesive.

[0069] Example 5:

[0070] This example prepares an albumin medical tissue adhesive, and the specific process is as follows:

[0071] (1) Weigh 25 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 25%. The mixed solution is slowly stirred at room temperature to form a uniform solution.

[0072] (2) Add 35 g of polyethylene glycol (PEG) to the above 25% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved.

[0073] (3) Cooling to room temperature to obtain rHSA-PEG adhesive.

[0074] Comparative Example 1:

[0075] This comparative example prepared a polyethylene glycol adhesive. The main difference from Example 1 is that no recombinant human albumin was added in this comparative example. The specific process is as follows:

[0076] (1) Weigh 60 g of polyethylene glycol (PEG) and dissolve it in 100 mL of aqueous solution to prepare a polyethylene glycol (PEG) aqueous solution with a mass volume concentration of 60%;

[0077] (2) The mixed solution is heated and stirred at a temperature below 60°C to form a homogeneous solution.

[0078] (3) Cool to room temperature to obtain PEG adhesive.

[0079] Comparative Example 2:

[0080] This comparative example prepared a polyethylene glycol adhesive. The main difference from Example 1 is that this comparative example uses blood-derived human albumin instead of recombinant human albumin. The specific process is as follows:

[0081] (1) Weigh 35 g of human serum albumin (pHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 35%. The mixed solution is slowly stirred at room temperature to form a uniform solution.

[0082] (2) Add 25 g of polyethylene glycol (PEG) to the above 35% pHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved.

[0083] (3) Cooling to room temperature to obtain pHSA-PEG adhesive.

[0084] Example 6: Tensile Strength and Elongation

[0085] 5 mL of each adhesive sample of Examples 1-5 and Comparative Examples 1-2 was applied to a stainless steel tank with a length, width, and depth of 10 cm × 1 cm × 0.5 cm. After solidification, the adhesive strip was carefully peeled off from the stainless steel tank and the tensile strength and elongation of the adhesive strip were measured using a universal testing machine. The results are shown in Table 1.

[0086] Table 1 Test results of adhesive tensile strength and elongation

[0087]

[0088] As can be seen from the results in Table 1, the tensile strength and elongation of the embodiment are significantly better than those of the control example, and the embodiment has good flexibility. In addition, the present invention further optimizes the dosage of the above components by using a compound of recombinant human albumin and polyethylene glycol, which can further improve the tensile strength and elongation of the adhesive.

[0089] Example 7: Adhesion Measurement

[0090] 5 mL of each adhesive sample from Examples 1-5 and Comparative Examples 1-2 was applied to a stainless steel circular groove with a diameter of 3 cm and a depth of 0.5 cm. A stainless steel disc with a diameter of 1 cm and a thickness of 0.1 cm was quickly placed on the adhesive. A drawstring was secured to the center of the other side of the disc. The groove and the disc drawstring were fixed to the fixtures on either side of a universal testing machine, and their maximum adhesion was measured. The results are shown in Table 2.

[0091] Table 2 Test results of maximum adhesive strength of adhesives

[0092]

[0093] As can be seen from the results in Table 2, the maximum adhesive force of the embodiment is significantly better than that of the comparative example. The cross-linking of recombinant human albumin and polyethylene glycol to form a gel helps to increase the viscosity of the adhesive, thereby improving the adhesion of the adhesive to the wound surface.

[0094] Example 8: Small Intestinal Anastomosis Adhesion Test in Rats

[0095] Forty-two SD rats were divided into seven groups, each consisting of six rats (half male and half female) and six rats (half male) treated with a cyanoacrylate group (α-butyl cyanoacrylate) and a test group (Examples 1-5, Comparative Examples 1-2). The rats were anesthetized with an intraperitoneal injection of 10% chloral hydrate (approximately 0.35 mL / 100 g). The small intestine was prepared using conventional intestinal preparation. The small intestine was cut with surgical scissors and then sutured with two stitches. 0.5 mL of adhesive was applied along the incision. A drainage tube was placed, the abdominal cavity was sutured, and the rats were kept and observed. The amount of drainage and the duration of drainage were recorded. Six weeks later, the rats were dissected to observe healing and pathological examination was performed. The results are shown in Table 3.

[0096] Table 3 Adhesion test of small intestine anastomosis in rats

[0097]

[0098]

[0099] Compared with the control group, the drainage time of Examples 1-5 was significantly reduced (P<0.01), and there was no intestinal adhesion after surgery, while there were 1 and 2 cases in the control group, respectively. There was no anastomotic leakage or anastomotic stenosis in the Example group, indicating that the Example group achieved a good therapeutic effect in intestinal anastomosis.

[0100] In summary, the adhesive of the present invention has a strong adhesive force, can significantly reduce restenosis and adhesion phenomena, has no toxic side effects, and has little irritation to the human body, and is suitable for adhesive repair of blood vessels.

[0101] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention, and such changes and modifications are intended to fall within the scope of the claims.

Claims

1. An albumin medical tissue adhesive, characterized in that: The adhesive comprises recombinant human albumin and polyethylene glycol, wherein the recombinant human albumin and polyethylene glycol are cross-linked by non-covalent interaction, wherein the non-covalent interaction is at least one of hydrogen bonding, hydrophobic interaction and van der Waals force. The recombinant human albumin is recombinant human blood albumin, recombinant human serum albumin or a mixture of the two obtained by genetic engineering technology; the genetic engineering technology refers to recombinant human albumin obtained by using Pichia pastoris, transgenic animals and plants heterologous expression system; The mass ratio of the recombinant human albumin to the polyethylene glycol is (25-35):(25-35); The recombinant human albumin contains free thiol groups whose proportion is not less than 80% of the total free radicals; The polyethylene glycol is one or a combination of PEG200, PEG400, PEG600, PEG800, PEG1000, PEG2000, and PEG4000; The method for using the albumin medical tissue adhesive comprises the following steps: (1) dissolving the recombinant human albumin to a concentration of 25-35% m / v at a dissolution temperature of less than 60° C. to obtain a recombinant human albumin solution; (2) adding the polyethylene glycol to the recombinant human albumin solution in step (1), wherein the temperature for dissolving the polyethylene glycol is less than 60° C.; (3) Cooling to obtain albumin medical tissue adhesive.

2. Use of the albumin medical tissue adhesive according to claim 1 in the preparation of medical repair materials or cosmetic repair materials.

3. A method for preparing an albumin medical tissue adhesive, characterized in that: Use any of the following methods: The first one: (1) Weigh 35 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 35%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution; (2) Add 25 g of polyethylene glycol (PEG) to the 35% rHSA aqueous solution in portions under heating and stirring at a temperature below 60°C until the PEG is completely dissolved; (3) cooling to room temperature to obtain rHSA-PEG adhesive; The second type: (1) Weigh 33 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 33%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution; (2) Add 27 g of polyethylene glycol (PEG) to the 33% rHSA aqueous solution in portions under heating and stirring at a temperature below 60°C until the PEG is completely dissolved; (3) cooling to room temperature to obtain rHSA-PEG adhesive; The third type: (1) Weigh 30 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare a rHSA aqueous solution with a mass volume concentration of 30%. The mixed solution was slowly stirred at room temperature to form a uniform solution; (2) Add 30 g of polyethylene glycol (PEG) to the 30% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved; (3) cooling to room temperature to obtain rHSA-PEG adhesive; The fourth type: (1) Weigh 27 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 27%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution; (2) Add 33 g of polyethylene glycol (PEG) to the 27% rHSA aqueous solution in portions under heating and stirring at a temperature below 60°C until the PEG is completely dissolved; (3) cooling to room temperature to obtain rHSA-PEG adhesive; Fifth type: (1) Weigh 25 g of recombinant human albumin (rHSA) and dissolve it in 100 mL of aqueous solution to prepare an rHSA aqueous solution with a mass volume concentration of 25%. The mixed solution was slowly stirred at room temperature to form a homogeneous solution; (2) Add 35 g of polyethylene glycol (PEG) to the 25% rHSA aqueous solution in portions under heating and stirring at a temperature below 60° C. until the PEG is completely dissolved; (3) cooling to room temperature to obtain rHSA-PEG adhesive; The recombinant human albumin is recombinant human blood albumin, recombinant human serum albumin or a mixture of the two obtained by genetic engineering technology; genetic engineering technology refers to recombinant human albumin obtained by using Pichia pastoris, transgenic animals and plants heterologous expression system; The recombinant human albumin contains free thiol groups whose proportion is not less than 80% of the total free radicals; The polyethylene glycol is one or a combination of PEG200, PEG400, PEG600, PEG800, PEG1000, PEG2000, and PEG4000.

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