US-20260124342-A1 - NANOSTRUCTURED LOW-IMMUNOGENIC BIOLOGICAL ARTIFICIAL BLOOD VESSEL AND PREPARATION METHOD THEREFOR

US20260124342A1US 20260124342 A1US20260124342 A1US 20260124342A1US-20260124342-A1

Abstract

A nanostructured low-immunogenic biological artificial blood vessel, a preparation method therefor and a use thereof. The preparation method comprises: decellularizing a pre-treated animal blood vessel to obtain a decellularized blood vessel; treating the decellularized blood vessel in an enzyme solution to obtain an enzyme-treated blood vessel, wherein the enzyme solution comprises nuclease and/or a biological enzyme; and using a cross-linking agent to cross-link the enzyme-treated blood vessel to obtain a nanostructured low-immunogenic biological artificial blood vessel, which is used as a blood vessel transplantation material. The artificial blood vessel overcomes the defects of decreased mechanical properties in decellularized biological tissues, in-vivo calcification after long-term use, and the presence of immunogenicity, antigenic components such as vascular wall cells and cell nuclei are fully removed, and original collagen and other extracellular matrix structural proteins in the tissue are retained to a greater extent, avoiding the in-vivo calcification of the blood vessel upon long-term implantation, enhancing the durability of use of the blood vessel; and the method has a short preparation cycle, low cost and high long-term patency rate.

Inventors

Xuefeng Qiu
Ying Guo
Bo Wang
Changdong ZHENG
Jianling SUN

Assignees

HUMATRIX MEDICAL TECHNOLOGY (SUZHOU) CO., LTD

Dates

Publication Date: 20260507
Application Date: 20231007
Priority Date: 20230728

Claims (12)

1 . A method for producing a biological artificial blood vessel having a nanostructure and low immunogenicity, comprising: S1) decellularizing a pretreated animal blood vessel to obtain a decellularized blood vessel, S2) treating the decellularized blood vessel with an enzyme solution to obtain an enzyme-treated blood vessel, wherein the enzyme solution comprises a nuclease and/or a biological enzyme, and S3) crosslinking the enzyme-treated blood vessel with a cross-linking agent to obtain the biological artificial blood vessel having a nanostructure and low immunogenicity.
2 . The method according to claim 1 , wherein the animal blood vessel is an artery or a vein of a large animal, wherein the large animal is selected from the group consisting of a pig, a sheep, a dog, a cow and a horse; the animal blood vessel is selected from the group consisting of an aorta, a pulmonary artery, a superior artery, an inferior artery, a common carotid artery, an internal jugular vein, an external jugular vein, a femoral artery, a femoral vein, an iliac artery, an iliac vein, a superior mesenteric artery, an inferior mesenteric artery, a rectal artery, a median sacral artery and a lower limb artery; the decellularization in step S1) is carried out using a decellularizing agent, the decellularizing agent comprises a detergent or a combination of a detergent and a chelating agent, wherein the detergent in the decellularizing agent has a concentration of 0.01 to 500 mmol/L, and the chelating agent in the decellularizing agent has a concentration of 0.01 to 500 mmol/L; and the decellularization is carried out at a temperature of 10° C. to 38° C. for 2 to 72 h, and the decellularizing agent is changed every 1 to 24 h during the decellularization.
3 . The method according to claim 2 , wherein in the case that the decellularizing agent comprises a detergent, the detergent is one or more selected from the group consisting of a non-ionic detergent, an anionic detergent and a cationic detergent, and an amphoteric detergent; in the case that the decellularizing agent comprises a combination of a detergent and a chelating agent, the detergent is one or more selected from the group consisting of a non-ionic detergent, an anionic detergent, a cationic detergent and an amphoteric detergent, wherein, the non-ionic detergent is one or more selected from the group consisting of polyethylene glycol, polyethylene glycol octylphenyl ether, polyol, polyoxyethylene fatty alcohol ether and polyoxyethylene alkyl phenol ether, the anionic detergent is one or more selected from the group consisting of sodium dodecyl sulfate, lithium dodecyl sulfate, sodium dodecyl sulfonate, sodium cholate and sodium deoxycholate, the cationic detergent is selected from the group consisting of benzalkonium bromide, cetyltrimethylammonium bromide and a combination thereof, the amphoteric detergent is selected from the group consisting of 3-[(3-cholamidopropyl)-dimethylammonium]-1-propanesulfonate inner salt, n-tetradecyl-N,N-dimethyl-3-ammonium-1-propanesulfonate and a combination thereof, and the chelating agent is selected from the group consisting of an inorganic chelating agent, an organic chelating agent and a combination thereof, wherein, the inorganic chelating agent is one or more selected from the group consisting of sodium tripolyphosphate, sodium hexametaphosphate and sodium pyrophosphate, the organic chelating agent is one or more selected from the group consisting of amino triacetic acid, ethylenediaminetetraacetic acid, ethylene glycol bis(tetraacetic acid), ethylenediamine diacetic acid, cyclohexane diaminetetraacetic acid, S,S-ethylenediamine disuccinic acid, diethyl triacetic acid, diethylenetriamine pentaacetic acid and a salt thereof, citric acid, tartaric acid, gluconic acid, hydroxyethyl ethylenediamine triacetic acid and dihydroxyethyl glycine; and the decellularizing agent further comprises sodium chloride, sodium hydroxide and water, wherein, the sodium chloride in the decellularizing agent has a concentration of 0.01 to 1.0 mmol/L, and the sodium hydroxide in the decellularizing agent has a concentration of 0.1 to 2.0 mmol/L.
4 . The method according to claim 1 , wherein the nuclease in the enzyme solution has a concentration of 1 to 50,000 KU/L, and/or the biological enzyme in the enzyme solution has a concentration of 1 to 50,000 KU/L, the enzyme solution further comprises 1 to 50 wt % human serum or animal serum, the enzyme solution further comprises physiological saline or buffer, and the enzyme treatment in step S2) is carried out at a temperature of 36° C. to 37° C. for 2 to 72 h, and the enzyme solution is changed every 1 to 24 h during the enzyme treatment.
5 . The method according to claim 1 , wherein after the enzyme treatment, step S2) further comprises treating the blood vessel with a solution comprising a biological enzyme to obtain an enzyme-treated blood vessel, the nuclease in the enzyme treatment is selected from the group consisting of DNase, RNase and a combination thereof, the biological enzyme in both the enzyme treatment and the solution comprising a biological enzyme is one or more selected from the group consisting of pepsin, lipase, trypsin, cathepsin, papain, ficain and subtilisin, the biological enzyme in the solution comprising a biological enzyme has a concentration of 1 to 50,000 KU/L, and the solution comprising a biological enzyme further comprises a solvent selected from the group consisting of physiological saline, buffer and a special solution for enzyme preparation, the treatment with the solution comprising a biological enzyme is carried out at a temperature of 36° C. to 37° C. for 0.25 to 48 h, and the solution comprising a biological enzyme is changed every 1 to 24 h during the treatment.
6 . The method according to claim 1 , wherein the cross-linking agent is one or more selected from the group consisting of a glutaraldehyde solution, an oxidized starch solution, a dialdehyde starch solution and a Jeffamine buffer, the glutaraldehyde solution has a concentration of 0.1 to 30 wt %, the oxidized starch solution has a concentration of 0.01 to 10 wt %, the dialdehyde starch solution has a concentration of 0.01 to 10 wt %, and the Jeffamine buffer has a concentration of 0.01 to 1.0 mol/L.
7 . The method according to claim 1 , wherein after the crosslinking, step S3) further comprises performing covalent binding of heparin to the cross-linked blood vessel by a chemical method to obtain the biological artificial blood vessel.
8 . The method according to claim 7 , wherein the covalent binding of heparin to the cross-linked blood vessel by a chemical method is carried out using a solution of carbodiimide, N-hydroxysulfosuccinimide and heparin in MES buffer, and the heparin in the solution for the covalent binding of heparin has a concentration of 1 to 200 mg/ml.
9 . A biological artificial blood vessel having a nanostructure and low immunogenicity produced by the method according to claim 1 , comprising a vascular collagen or a nanofibrous structure formed by vascular collagen and elastin.
10 . The biological artificial blood vessel having a nanostructure and low immunogenicity according to claim 9 , wherein the vascular collagen includes type I collagen, type III collagen and/or type IV collagen, in the case that the biological artificial blood vessel comprises only the vascular collagen, the vascular collagen has a content of 30% to 85% of the dry weight of the biological artificial blood vessel, and in the case that the biological artificial blood vessel comprises the vascular collagen and elastin, the vascular collagen has a content of 20% to 55% of the dry weight of the biological artificial blood vessel, and the elastin has a content of 20% to 55% of the dry weight of the biological artificial blood vessel.
11 . Use of the biological artificial blood vessel having a nanostructure and low immunogenicity produced by the method according to claim 1 in the manufacture of a vascular graft material for hemodialysis vascular access in chronic renal failure, a vascular graft material for arterial trauma of lower limbs, a vascular graft material for peripheral artery bypass grafting or a vascular graft material for coronary artery bypass grafting.
12 . Use of the biological artificial blood vessel having a nanostructure and low immunogenicity produced by the method according to claim 1 in the manufacture of a vascular graft material for vascular access of pateints with chronic haemodialysis; wherein, the biological artificial blood vessel is used as a replacement repair graft material for failure, infection or aneurys formation after chronic dialysis autologous arteriovenous fistula creation or artificial vascular fistula creation.

Description

This application claims the priority of Chinese Patent Application No. 202310942899.8, filed with the China National Intellectual Property Administration on Jul. 28, 2023, and titled with “NANOSTRUCTURED LOW-IMMUNOGENIC BIOLOGICAL ARTIFICIAL BLOOD VESSEL AND PREPARATION METHOD THEREFOR”, the disclosure of which is hereby incorporated by reference in its entirety. FIELD The present disclosure relates to the technical field of medical devices, in particular to a biological artificial blood vessel having a nanostructure and low immunogenicity and a production method thereof. BACKGROUND Nowadays, according to the statistics of the World Health Organization, the prevalence of cardiovascular diseases is increasing with the development of society, which have become one of the killers endangering human health and the leading cause of death among humans. According to the statistics, there are currently more than 11 million coronary heart disease cases in China, with more than 5 million patients requiring revascularization. The number of coronary artery bypass grafting surgeries in China reached nearly 100,000 in 2022, a ten-fold increase from 10 years ago. In 2019, there were approximately 45.3 million patients with peripheral atherosclerosis in China, with about 8% of patients requiring revascularization. However, the current penetration rate of surgery, including the interventional surgery, is only 0.3%, indicating a huge market space. China National Renal Data System (CNRDS) shows that the hemodialysis rate is increasing year by year (increasing by 13% to 14%/year). In China, there are currently over 130 million patients with chronic renal insufficiency, with more than 3 million patients requiring hemodialysis. The number of registered patients undergoing dialysis was about 200,000 in 2011, 630,000 in 2019, 690,000 in 2020 and 750,000 in 2021, and it is estimated that the number will be about 1.27 million in 2027 and more than 1.6 million in 2030. Among them, more than 80% of the patients received arteriovenous fistula creation by conventional surgery, 12% received central venous catheters, and about 7.2% received artificial vascular access (CNRDS data published at the 6th Summit Forum on Nephrology, Renal Intervention, and Blood Purification Devices held in Pengcheng in November 2022). In the United States, there are about 650,000 patients undergoing long-term chronic haemodialysis per year. Among them, about 65% of the patients received arteriovenous fistula creation by conventional surgery, 15% received central venous catheters, and about 20% received artificial vascular access. After the creation of an arteriovenous fistula by conventional surgery, it is necessary for patients to wait 3 to 6 months for the fistula to mature before it can be used as a dialysis access. In China, one-third to one-half of patients may experience failed fistula maturation 3-6 months after the surgical creation of the arteriovenous fistula, and thus cannot receive dialysis. Therefore, only small-caliber artificial blood vessels can be used, but this access is prone to aneurysms formation and then deterioration. Central venous catheters can only be used as a temporary dialysis access, which shows an annual infection rate of 200%, many complications and a high mortality in patients. Traditional expanded polytetrafluoroethylene (ePTFE) artificial blood vessels are susceptible to infection and formation of intraluminal thrombosis, with a 1-year patency rate of about 10-30% and a service life of about 2 years, severely limiting clinical outcomes. Therefore, the development of small-caliber vessels suitable for coronary artery disease, lower limb artery replacement, and haemodialysis vascular access in patients with end-stage renal disease is of great clinical significance. Vascular repair and replacement is one of the most effective options for the treatment of vascular diseases. Currently, blood vessel sources generally include autologous blood vessels, artificial blood vessels made of polymer materials and foreign blood vessels, and tissue-engineered blood vessels are in the research and development stage. Autologous internal mammary artery or radial artery shows an excellent medium and long-term patency rate, and the coronary artery bypass grafting thereof has a 10-year patency rate of about 90% or more. Autologous great saphenous vein shows a 5-year patency rate of about 60-70% and a 10-year patency rate of about 40-50%, and their calibers are only matched for coronary artery bypass grafting and not for other clinical indications, for which reason they cannot be used in other clinical indications. In addition, the source of autologous blood vessels is limited, and 10-30% of patients cannot provide great saphenous veins due to varicose great saphenous veins or obesity, diabetes and other factors. The harvest of saphenous vein may cause secondary trauma and complications such as incision infection, lower limb edema, and prolongation o