Gutta-percha-based bionic blood vessel and preparation method thereof

Abstract

The invention provides a gutta-percha-based bionic blood vessel and a preparation method thereof, and belongs to the technical field of biomedical materials. By utilizing the unique rubber and plastic duality of the natural eucommia rubber, especially excellent biocompatibility and mechanical compliance, the problems of low bursting pressure of a synthetic material artificial blood vessel and poor compliance of a natural material bionic blood vessel are solved. The result of the embodiment shows that the blood compliance of the gutta-percha-based bionic blood vessel is 6.5%-20% under the blood pressure of 80 mmHg to 120 mmHg, the bursting pressure is 4500 mmHg to 8000 mmHg, the blood compliance and the bursting pressure of the gutta-percha-based bionic blood vessel are higher than those of arterial blood vessels of the human body, and the use requirements of the human body are met.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedical materials, and in particular relates to an eucommia gum-based bionic blood vessel and a preparation method thereof. Background Art

[0002] Cardiovascular disease (CVD) is one of the leading causes of death worldwide. According to the World Health Organization's Global Health Survey, about 1.5% of the world's population suffers from cardiovascular disease. In recent years, with the aging of my country's population and the impact of unhealthy lifestyles, the incidence and mortality of CVD patients have also increased year by year. It is estimated that the number of CVD patients in my country has reached 330 million, and it ranks first in the proportion of disease deaths among residents.

[0003] Artificial blood vessels or bionic blood vessels are materials used to replace or repair damaged or defective blood vessels. They can be used as blood vessel substitutes or stents to restore blood flow and maintain normal blood circulation in the body. middle Small Three specifications. According to the material properties, they can be divided into synthetic material blood vessels and biomaterial blood vessels. Synthetic materials include polyvinyl alcohol (PVA), polyurethane (PU), polytetrafluoroethylene (PTFE), polycaprolactone (PCL), polyethylene terephthalate (PET), etc. These materials are processed into artificial blood vessels by knitting, weaving, braiding or non-woven methods. They have certain mechanical strength and durability, but their biocompatibility is relatively poor, which is specifically manifested in the relatively small bursting pressure. For example, the bursting pressure of small-caliber PU-PGACL and PCL / PLCL artificial blood vessels does not exceed 2500mmHg, and the bursting pressure of PCL-NFC / PVA-NPs artificial blood vessels does not exceed 1800mmHg. Biomaterials include polylactic acid ester (PLA), protein and fiber, etc., which are not very flexible, specifically manifested in low blood compliance. For example, the blood compliance of collagen / PLA composite bionic blood vessels reported in the literature does not exceed 4% at 100mmHg; the blood compliance of PVA and its silk nanofiber composite bionic blood vessels does not exceed 1.5% at 100mmHg; the blood compliance of fiber textile bionic blood vessels does not exceed 1.2% at normal blood pressure (80-120mmHg), and the bursting pressure does not exceed 1000mmHg; the blood compliance of micro-nano fiber composite bionic blood vessels does not exceed 4.5% at normal blood pressure. The blood compliance of human venous blood vessels is 0.7-1.5%, and the bursting pressure is 1600-2300mmHg; the blood compliance of arterial blood vessels is 4.6-6.2%, and the bursting pressure is 2000-4300mmHg. It can be seen that the bursting pressure and blood compliance of current bionic blood vessels cannot meet the use needs of the human body. Summary of the invention

[0004] The purpose of the present invention is to provide a eucommia gum-based bionic blood vessel and a preparation method thereof. The eucommia gum-based bionic blood vessel prepared by the preparation method provided by the present invention has high blood compliance and high bursting pressure.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a method for preparing an eucommia gum-based bionic blood vessel, comprising the following steps:

[0007] (1) dissolving and mixing natural eucommia rubber, biopolymer and an organic solvent to obtain a mixed solution;

[0008] (2) sticking a rod-shaped mold in the mixed solution obtained in step (1) and then vertically pulling it out to cool and shape it, thereby obtaining a mold for sticking the material;

[0009] (3) demolding the mold of the adhesion material obtained in step (2) to obtain eucommia gum-based bionic blood vessels.

[0010] Preferably, the biopolymer in step (1) is any one of polyvalerolactone, polycaprolactone, alkyl ester and olefin ester of biomass material; the biomass material includes any one of cellulose, hemicellulose, lignin, starch, pectin, chitosan, sodium alginate, protein, nucleic acid, saponin and rosin; the carbon content of the alkyl ester is 12 to 24; the carbon content of the olefin ester is 12 to 24.

[0011] Preferably, in step (1), the mass ratio of the biopolymer to the natural eucommia rubber is (0-1000):1.

[0012] Preferably, the material of the rod-shaped mold in step (2) is any one of glass, metal, silicone, wood, plastic and resin; the surface roughness R of the rod-shaped mold is a 0.1~300nm.

[0013] Preferably, the adhesion-cooling molding operation in step (2) is repeated 1 to 10 times.

[0014] Preferably, the organic solvent in step (1) comprises chloroform, dichloromethane, petroleum ether, toluene, tetrahydrofuran, acetone or ethyl acetate.

[0015] Preferably, the viscosity of the mixed solution in step (1) is 0.01 to 100 Pa·s.

[0016] Preferably, the temperature for dissolving and mixing in step (1) is 60 to 120° C., and the time is 2 to 6 hours.

[0017] Preferably, the temperature of the adhesion in step (2) is 50-100° C. and the time is 1-30 min.

[0018] The present invention also provides an eucommia gum-based bionic blood vessel prepared by the preparation method described in the above technical solution.

[0019] The present invention provides a method for preparing an eucommia gum-based bionic blood vessel, comprising the following steps: (1) dissolving and mixing natural eucommia rubber, biopolymer and an organic solvent to obtain a mixed solution; (2) adhering a rod-shaped mold in the mixed solution obtained in the step (1) and then vertically withdrawing and cooling the mold to obtain a mold of the adhered material; (3) demoulding the mold of the adhered material obtained in the step (2) to obtain an eucommia gum-based bionic blood vessel. The present invention utilizes the unique rubber-plastic duality of natural eucommia rubber, especially the excellent biocompatibility and mechanical compliance, to overcome the problems of low bursting pressure of synthetic artificial blood vessels and poor compliance of natural material bionic blood vessels. The results of the embodiment show that the blood compliance of the eucommia gum-based bionic blood vessel provided by the present invention is 6.5-20% at a blood pressure of 80-120 mmHg, and the bursting pressure is 4500-8000 mmHg, which is higher than the blood compliance and bursting pressure of human arterial blood vessels and meets the use requirements of the human body. DETAILED DESCRIPTION

[0020] All raw materials of the present invention have no particular limitation on their sources and can be purchased from the market or prepared according to conventional methods known to those skilled in the art.

[0021] There is no particular limitation on the purity of all raw materials in the present invention. The present invention preferably uses analytically pure raw materials or raw materials with a purity commonly used in the field of biomedical materials.

[0022] The present invention provides a method for preparing an eucommia gum-based bionic blood vessel, comprising the following steps:

[0023] (1) dissolving and mixing natural eucommia rubber, biopolymer and an organic solvent to obtain a mixed solution;

[0024] (2) sticking a rod-shaped mold in the mixed solution obtained in step (1) and then vertically pulling it out to cool and shape it, thereby obtaining a mold for sticking the material;

[0025] (3) demolding the mold of the adhesion material obtained in step (2) to obtain eucommia gum-based bionic blood vessels.

[0026] The invention dissolves and mixes natural eucommia rubber, biopolymer and organic solvent to obtain a mixed solution.

[0027] Natural eucommia rubber has unique rubber-plastic duality, especially excellent biocompatibility and mechanical compliance. The present invention uses natural eucommia rubber as the material of bionic blood vessels, overcoming the problems of low bursting pressure of synthetic artificial blood vessels and poor compliance of natural bionic blood vessels.

[0028] In the present invention, the biopolymer is preferably any one of polyvalerolactone, polycaprolactone, alkyl ester and olefin ester of biomass material; the biomass material preferably includes any one of cellulose, hemicellulose, lignin, starch, pectin, chitosan, sodium alginate, protein, nucleic acid, saponin and rosin; the carbon number of the alkyl ester is preferably 12 to 24; the carbon number of the olefin ester is preferably 12 to 24. In an embodiment of the present invention, the biopolymer can be cellulose polyvalerolactone; it can also be starch dodecyl ester; it can also be lignin polycaprolactone; it can also be rosin tetracosene (C2:1) ester. The above biopolymer can be cross-linked and compounded with natural eucommia rubber to further improve the bursting pressure of the bionic blood vessel.

[0029] In the present invention, the mass ratio of the biopolymer to the natural eucommia rubber is (0-1000):1, preferably (1-100):1. As an embodiment of the present invention, the mass ratio of the biopolymer to the natural eucommia rubber can be 1:1, 5:1, 10:1, 50:1, 100:1, 200:1, 300:1, 400:1, 500:1, 600:1 or 1000:1. The mass ratio of the biopolymer to the natural eucommia rubber within the above range is conducive to further improving the burst pressure of the bionic blood vessel while avoiding excessive decrease in blood compliance.

[0030] In the present invention, the organic solvent preferably includes chloroform, dichloromethane, petroleum ether, toluene, tetrahydrofuran, acetone or ethyl acetate. The above organic solvents have good solubility for natural eucommia rubber and biopolymers and are easy to remove.

[0031] In the present invention, the dissolving and mixing temperature is preferably 60-120° C., more preferably 80-100° C.; as an embodiment of the present invention, the dissolving and mixing temperature may be 65° C., 75° C., 85° C., 90° C., 95° C., 105° C. or 110° C. Within the above temperature range, the dissolving speed of natural eucommia rubber and biopolymer can be increased.

[0032] In the present invention, the dissolution and mixing time is preferably 2 to 6 hours, more preferably 3 to 5 hours; as an embodiment of the present invention, the dissolution and mixing time can be 2.5 hours, 3.5 hours, 4 hours, 4.5 hours or 5.5 hours. When the dissolution and mixing time is within the above range, the natural eucommia rubber and the biopolymer can be completely dissolved.

[0033] In the present invention, the viscosity of the mixed liquid is preferably 0.01 to 100 Pa·s, more preferably 0.1 to 10 Pa·s, and further preferably 0.5 to 5 Pa·s. The viscosity of the mixed liquid within the above range is conducive to the adhesion of the mixed liquid to the mold.

[0034] As an embodiment of the present invention, the viscosity of the mixed solution can be adjusted by the amount of solvent added, and the mass volume ratio of the biopolymer material to the organic solvent can be 1g: (0.5-1000)mL.

[0035] After the mixed liquid is obtained, the present invention adheres the rod-shaped mold in the mixed liquid and then vertically draws it out for cooling and forming to obtain a mold for adhering the material.

[0036] In the present invention, the material of the rod-shaped mold is preferably any one of glass, metal, silicone, wood, plastic and resin; the surface roughness R of the rod-shaped mold is a Preferably, it is 0.1 to 300 nm, more preferably 1 to 50 nm; As one embodiment of the present invention, the surface roughness R of the rod-shaped mold is a It can be 0.5nm, 1nm, 5nm, 10nm, 20nm, 50nm, 100nm or 200nm. Material and surface roughness R of the rod mold a Within the above range, it is beneficial to the adhesion and demoulding of the material.

[0037] The present invention has no special requirements on the size of the rod-shaped mold, and can be adjusted accordingly according to the specifications of the bionic blood vessel; as an embodiment of the present invention, the diameter of the rod-shaped mold can be 0.1 to 50 mm, or 0.5 to 20 mm or 1 to 10 mm.

[0038] In the present invention, the adhesion temperature is preferably 50-100° C., more preferably 60-70° C.; as an embodiment of the present invention, the adhesion temperature may be 55° C., 65° C., 75° C., 85° C., 90° C. or 95° C. Within the above temperature range, the adhesion of the mixed solution is favorable.

[0039] In the present invention, the adhesion time is preferably 1 to 30 minutes, more preferably 2 to 15 minutes, and further preferably 3 to 8 minutes; as an embodiment of the present invention, the adhesion time can be 4 minutes, 5 minutes, 10 minutes, 12 minutes or 25 minutes. When the adhesion time is within the above range, both adhesion effect and production efficiency can be taken into account.

[0040] In the present invention, the step of adhesion-cooling molding is preferably repeated 1 to 10 times, more preferably 4 to 6 times; as an embodiment of the present invention, the step of adhesion-cooling molding can be repeated 2 times, 3 times, 5 times, 7 times, 8 times or 9 times. Repeating adhesion-cooling molding is conducive to further improving the mechanical properties and thickness of the bionic blood vessel, and the number of repetitions can be adjusted according to different usage requirements.

[0041] The present invention has no particular limitation on the specific operation of the adhesion, and the conventional operation method in the art can be used. In the embodiment of the present invention, the rod-shaped mold is vertically inserted into the mixed liquid, and after adhesion, it is vertically pulled out.

[0042] In the present invention, the cooling molding temperature is preferably 10-30° C., more preferably 15-25° C.; as an embodiment of the present invention, the cooling molding temperature can be 12° C., 14° C., 18° C., 20° C., 23° C. or 26° C. Within the above temperature range, the molding of bionic blood vessels is beneficial.

[0043] In the present invention, the cooling molding time is preferably 0.5 to 5 minutes, more preferably 1 to 3 minutes; as an embodiment of the present invention, the cooling molding time can be 1 minute, 1.5 minutes, 2 minutes, 3 minutes or 4 minutes. When the cooling molding time is within the above range, the bionic blood vessel can be fully cooled.

[0044] The method demoulds the obtained adhesion material from the mold to obtain the eucommia gum-based bionic blood vessel.

[0045] In the present invention, the organic solvent is preferably completely volatilized before demoulding. The present invention has no special requirements on the method of volatilization of the organic solvent, and a conventional solvent volatilization method can be adopted; as an embodiment of the present invention, the organic solvent can be completely volatilized under room temperature and ventilation conditions.

[0046] The present invention has no special requirements for the specific demoulding operation, and conventional demoulding operations in the art can be used.

[0047] The present invention utilizes the unique rubber-plastic duality of natural eucommia rubber, especially the excellent biocompatibility and mechanical compliance, to overcome the problems of low bursting pressure of synthetic artificial blood vessels and poor compliance of natural bionic blood vessels; by adding other biopolymers, the bursting pressure of the bionic blood vessels can be further improved to meet the use requirements of different parts; in addition, the bionic blood vessels provided by the present invention also have good ductility and excellent biocompatibility, which expands the adaptability of the bionic blood vessels; the preparation method provided by the present invention is simple and easy to operate, low in cost, and is conducive to industrial production.

[0048] The present invention also provides eucommia gum-based bionic blood vessels prepared by the preparation method described in the above technical solution.

[0049] The present invention has no particular limitation on the length and thickness of the Eucommia gum-based bionic blood vessels, which can be conventionally adjusted according to actual application requirements.

[0050] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0051] Example 1

[0052] A method for preparing an eucommia gum-based bionic blood vessel, comprising the following specific steps:

[0053] (1) Dissolve 1 g of natural eucommia rubber in 0.5 mL of chloroform at 60°C for 2 h to form a mixed solution with a viscosity of 0.01 Pa·s;

[0054] (2) inserting a glass rod mold with a surface roughness of 0.1 nm and a diameter of 0.1 mm vertically into the mixed solution formed in step (1), adhering to the mixture for 1 min, and then vertically withdrawing the mold, cooling at 10° C. for 0.5 min to form the mold; repeating the operation once, during which the mixed solution was kept warm at 50° C.; and obtaining a mold for adhering the material;

[0055] (3) The mold of the adhesion material obtained in step (2) is placed under room temperature and ventilation conditions to evaporate the chloroform, and then demoulding is performed to obtain an eucommia gum-based bionic blood vessel.

[0056] Example 2

[0057] A method for preparing an eucommia gum-based bionic blood vessel, comprising the following specific steps:

[0058] (1) Dissolve 1 g of natural eucommia rubber and 1000 g of cellulose polyvalerolactone in 1000 mL of dichloromethane at 120°C for 6 h to form a mixed solution with a viscosity of 100 Pa·s;

[0059] (2) inserting a stainless steel rod mold with a surface roughness of 300 nm and a diameter of 50 mm vertically into the mixed liquid formed in step (1), adhering for 30 minutes, and then vertically withdrawing it, cooling it at 30° C. for 5 minutes to form it; repeating the operation 10 times, during which the mixed liquid was kept warm at 100° C.; and obtaining a mold for adhering the material;

[0060] (3) The mold of the adhesion material obtained in step (2) is placed under room temperature and ventilation conditions to evaporate the dichloromethane, and then demolded to obtain an eucommia gum-based bionic blood vessel.

[0061] Example 3

[0062] A method for preparing an eucommia gum-based bionic blood vessel, comprising the following specific steps:

[0063] (1) Dissolve 1 g of natural eucommia rubber and 10 g of lignin polycaprolactone in 500 mL of toluene at 80 °C for 3 h to form a mixed solution with a viscosity of 50 Pa·s;

[0064] (2) inserting a stainless steel rod mold with a surface roughness of 100 nm and a diameter of 10 mm vertically into the mixed solution formed in step (1), adhering for 5 minutes, and then vertically withdrawing it, cooling it at 20° C. for 2 minutes to form it; repeating the operation twice, during which the mixed solution was kept warm at 60° C.; and obtaining a mold for adhering the material;

[0065] (3) The mold of the adhesive material obtained in step (2) is placed under room temperature and ventilation conditions to evaporate the toluene, and then demoulding is performed to obtain an eucommia gum-based bionic blood vessel.

[0066] Example 4

[0067] A method for preparing an eucommia gum-based bionic blood vessel, comprising the following specific steps:

[0068] (1) Dissolve 1 g of natural eucommia rubber and 100 g of starch laurate in 800 mL of petroleum ether at 90° C. for 4 h to form a mixed solution with a viscosity of 80 Pa·s;

[0069] (2) inserting a silicone rod mold with a surface roughness of 200 nm and a diameter of 20 mm vertically into the mixed solution formed in step (1), adhering for 15 minutes, then vertically withdrawing, and cooling at 25° C. for 3 minutes to form; repeating the operation 5 times, during which the mixed solution was kept warm at 70° C.; and obtaining a mold for the adhered material;

[0070] (3) The petroleum ether is completely evaporated from the mold of the adhesive material obtained in step (2) under ventilation conditions at room temperature, and the mold is demoulded to obtain an eucommia gum-based bionic blood vessel.

[0071] Example 5

[0072] A method for preparing an eucommia gum-based bionic blood vessel, comprising the following specific steps:

[0073] (1) Dissolve 1 g of natural eucommia rubber and 50 g of rosin tetracosene (C2:1) ester in 300 mL of acetone at 100 °C for 5 h to form a mixed solution with a viscosity of 60 Pa·s;

[0074] (2) inserting a wood stick mold with a surface roughness of 50 nm and a diameter of 30 mm vertically into the mixed solution formed in step (1), adhering for 20 minutes, and then vertically withdrawing it, cooling it at 15° C. for 2 minutes to form it; repeating the operation 8 times, during which the mixed solution was kept warm at 90° C.; and obtaining a mold for the adhered material;

[0075] (3) The mold of the adhesive material obtained in step (2) is placed under room temperature and ventilation conditions to evaporate the acetone, and then demoulding is performed to obtain an eucommia gum-based bionic blood vessel.

[0076] Test Example 1

[0077] According to GB / T 528-2019 and the international standard ANSI 7198, the elongation at break, blood compliance and bursting pressure of the eucommia gum-based bionic blood vessels prepared in each embodiment were tested respectively, and the test results are shown in Table 1.

[0078] Table 1 Test results of the performance of eucommia gum-based bionic blood vessels prepared in various embodiments

[0079] Elongation at break Blood compliance at 80-120 mmHg 80~120mmHg burst pressure Example 1 600％ 20％ 4500mmHg Example 2 180％ 6.5％ 8000mmHg Example 3 230％ 8％ 5820mmHg Example 4 350％ 10％ 6580mmHg Example 5 650％ 15％ 7530mmHg

[0080] According to the test results in Table 1, it can be seen that the bionic blood vessel prepared using pure natural eucommia rubber has the highest blood compliance at a normal blood pressure of 80-120 mmHg, reaching 20%, but the bursting pressure is also the lowest, only 4500 mmHg. By adding other biopolymers, the bursting pressure of the bionic blood vessel increases, but the blood compliance decreases. In Example 2, the mass ratio of natural eucommia rubber to other biopolymers is 1:1000, and the blood compliance is only 6.5%, which is close to the blood compliance of human arteries.

[0081] Test Example 2

[0082] The biocompatibility of the Eucommia gum-based bionic blood vessels prepared in Example 1 was tested:

[0083] L929 cells with good growth conditions were digested with 0.25% trypsin for 2 min;

[0084] L929 cells were washed twice with 10% FCS-RPMI-1640 culture medium to remove the digestion solution and the original growth medium;

[0085] The L929 cell concentration was adjusted to 2×10 5 / mL;

[0086] Add L929 cell suspension to a 35 mm culture dish;

[0087] Cut the sample to be tested and cover the cells in the culture dish, and set up a culture medium blank control;

[0088] Place in a 37°C, 5% CO2 incubator for 56 h;

[0089] The culture dish was removed and MTT, 1 mL, was added;

[0090] Place in a 37°C, 5% CO2 incubator and continue culturing for 4 h;

[0091] Take out the culture dish, add 10 mL of acidified isopropanol, and place at room temperature for 10 to 20 minutes. Pipette and shake to mix thoroughly to fully dissolve the MTT-formazan product.

[0092] The OD values ​​were measured using an enzyme-labeled instrument with a detection wavelength of 570 nm and a reference wavelength of 630 nm.

[0093] The survival rate of mouse fibroblast L929 in the eucommia gum-based bionic blood vessels provided in Example 1 can be as high as over 98%, indicating that the eucommia gum-based bionic blood vessels provided by the present invention have excellent biocompatibility.

[0094] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing an eucommia gum-based bionic blood vessel, characterized in that: The steps include: (1) dissolving and mixing natural eucommia rubber, biopolymer and an organic solvent to obtain a mixed solution; (2) sticking a rod-shaped mold in the mixed solution obtained in step (1) and then vertically pulling it out to cool and shape it, thereby obtaining a mold for sticking the material; (3) demolding the mold of the adhesion material obtained in step (2) to obtain eucommia gum-based bionic blood vessels.

2. The preparation method according to claim 1, characterized in that: In the step (1), the biopolymer is any one of polyvalerolactone, polycaprolactone, alkyl ester and olefin ester of the biomass material; the biomass material includes any one of plant cellulose, hemicellulose, lignin, starch, pectin, chitosan, sodium alginate, protein, nucleic acid, saponin and rosin; the carbon number of the alkyl ester is 12 to 24; the carbon number of the olefin ester is 12 to 24.

3. The preparation method according to claim 1 or 2, characterized in that: In the step (1), the mass ratio of the biopolymer to the natural eucommia rubber is (0-1000):

1.

4. The preparation method according to claim 1, characterized in that: The material of the rod-shaped mold in step (2) is any one of glass, metal, silicone, wood, plastic and resin; the surface roughness R of the rod-shaped mold is a 0.1~300nm.

5. The preparation method according to claim 1, characterized in that: The adhesion-cooling molding operation in step (2) is repeated 1 to 10 times.

6. The preparation method according to claim 1, characterized in that: The organic solvent in step (1) includes chloroform, dichloromethane, petroleum ether, toluene, tetrahydrofuran, acetone or ethyl acetate.

7. The preparation method according to claim 1, characterized in that: The viscosity of the mixed solution in step (1) is 0.01 to 100 Pa·s.

8. The preparation method according to claim 1, characterized in that: The temperature for dissolving and mixing in step (1) is 60 to 120° C. and the time is 2 to 6 hours.

9. The preparation method according to claim 1, characterized in that: The temperature of the adhesion in the step (2) is 50 to 100° C. and the time is 1 to 30 minutes.

10. Eucommia gum-based bionic blood vessels prepared by the preparation method according to any one of claims 1 to 9.