Blood stabilizer, blood bag and preparation method of blood stabilizer

Abstract

The invention discloses a blood stabilizer, a blood bag and a preparation method of the blood stabilizer, the blood stabilizer is a polymeric monomer or an oligomer of the polymeric monomer, and the polymeric monomer is composed of ethylene glycol, propylene glycol and glycerol according to the molar ratio of 1: 1: (0.5-2). The invention discloses a blood stabilizer blood bag. The blood bag is made of a modified polyolefin material. The preparation method of the blood stabilizer blood bag comprises the following preparation steps: S1, mixing the raw materials; s2, carrying out melt blending; and S3, film blowing and bag making. Through the synergistic effect of the three-dimensional hydrogen bond network stabilizer and the modified polyolefin material, the preservation period of the red blood cells is prolonged to 45 days from traditional 35 days, a dynamic protection film formed by the stabilizer can inhibit a glycolytic pathway, the 2, 3-DPG content retention rate reaches 85% or above, compared with the prior art, the 2, 3-DPG content retention rate is increased by 15%, the oxygen carrying function of the red blood cells is effectively maintained, and the service life of the red blood cells is prolonged. And after being stored for 45 days, the red blood cell survival rate is still greater than or equal to 92%, the hemolysis rate is less than or equal to 0.05%, and the clinical long-term blood use

Description

Technical Field

[0001] The present invention relates to the technical field of blood stabilizers, and particularly to a blood stabilizer, a blood bag and a preparation method thereof. Background Art

[0002] Currently, the blood preservation systems used clinically mainly rely on the synergistic effect of polyvinyl chloride (PVC) blood bags and anticoagulants. Since traditional PVC blood bags add di-(2-ethylhexyl) phthalate (DEHP) as a plasticizer, there are serious safety hazards: after DEHP migrates into the blood, it can cause hepatotoxicity, reproductive toxicity and endocrine disruption. In addition, the surface of the PVC material has strong hydrophobicity (water contact angle > 90°), resulting in a platelet adhesion rate as high as 15%-20%, activating the coagulation system and shortening the red blood cell preservation period, usually ≤ 35 days. Although the patent with the publication number "CN104403122A" proposes to use a poly(lactic-co-glycolic acid) (PLGA) coating to improve compatibility, the binding force between the coating and the PVC substrate is weak, less than 2 N / cm, and it is easy to fall off during long-term storage, resulting in the hemolysis rate increasing to more than 0.3%.

[0003] And to solve the toxicity problem of DEHP, existing technologies such as patent CN104403122A turn to polyolefin materials, but there are still three major defects: 1. The polyolefin surface lacks active groups, and the complement activation rate > 12% when in direct contact with blood; 2. The antibacterial performance is insufficient, and antibacterial agents such as silver ions need to be added additionally, increasing the risk of blood contamination; 3. The process is complex, and the layer-by-layer self-assembly technology used requires multiple coatings, increasing the production cost by more than 40%. In addition, existing stabilizers are mostly small molecule alcohols such as propylene glycol, and their critical micelle concentration (CMC) > 0.5 mmol / L, which cannot form a stable protective film at the blood interface, resulting in the red blood cell preservation period being only extended to 38 days. Summary of the Invention

[0004] The purpose of the present invention is to solve the disadvantages existing in the prior art, and to propose a blood stabilizer, a blood bag and a preparation method thereof.

[0005] In order to achieve the above purpose, the present invention adopts the following technical scheme:

[0006] A blood stabilizer, the stabilizer is a polymer monomer or its oligomer, the polymer monomer is composed of ethylene glycol, propylene glycol and glycerol in a molar ratio of 1:1:0.5-2; the molecular weight of the stabilizer is 100-5000 Da, and a three-dimensional network structure is formed through intermolecular hydrogen bonds.

[0007] Preferably, the stabilizer is prepared by free radical polymerization reaction under the following reaction conditions: ammonium persulfate is used as an initiator, and the ammonium persulfate accounts for 1-3% of the total mass of the polymerization monomers, and the reaction is carried out at 60-80 °C for 2-6 hours; ethylene glycol diglycidyl ether is added as a crosslinking agent during the reaction process to control the molecular weight distribution, and the ethylene glycol diglycidyl ether accounts for 0.5-2% of the total mass of the monomers.

[0008] Preferably, the stabilizer contains a quaternary ammonium salt modifier, and the quaternary ammonium salt modifier is cetyltrimethylammonium bromide, and the addition amount is 1-2% of the total mass of the polymerization monomers.

[0009] A blood stabilizer blood bag, the blood bag is made of a modified polyolefin material, and the surface of the modified polyolefin material is grafted with the blood stabilizer as described in any one of claims 1-3 to form a coating with a thickness of 0.1-5 μm; the water contact angle of the blood bag is ≤60°, the platelet adhesion rate is ≤3%, and the hemolysis rate is ≤0.05%.

[0010] Preferably, the modified polyolefin material is a polyethylene-polypropylene blend, which is prepared by the following steps:

[0011] a. Under an argon atmosphere, the surface of the film is treated with a plasma processor (power 100-300 W) for 1-3 hours;

[0012] b. Graft the blood stabilizer as described in any one of claims 1-3, the treatment temperature is 30-60 °C, and the treatment time is 2-4 hours;

[0013] c. After treatment, a C-O-C bonding structure is formed on the surface of the film, and the bonding strength is ≥5 N / cm.

[0014] A method for preparing a blood stabilizer blood bag, comprising the following preparation steps:

[0015] S1. Raw material mixing, mixing the blood stabilizer as described in any one of claims 1-3 and the modified polyolefin raw material in a mass ratio of 1:100-1:500;

[0016] S2. Melt blending, melt blending with a twin-screw extruder under the conditions of a temperature of 180-200 °C and a rotation speed of 50-80 revolutions per minute;

[0017] S3. Film blowing and bag making, blowing the melt-blended raw materials into a film with a thickness of 50-100 μm, using the film for blood bag making, and the heat sealing strength after bag making is ≥15 N / 15 mm.

[0018] The present invention has the following beneficial effects:

[0019] 1. Through the synergistic effect of the three-dimensional hydrogen bond network stabilizer and the modified polyolefin material, the present invention extends the red blood cell storage period from the traditional 35 days to 45 days. The dynamic protective film formed by the stabilizer can inhibit the glycolytic pathway, enabling the retention rate of 2,3-DPG content to reach over 85%, a 15% increase compared to the prior art, effectively maintaining the oxygen-carrying function of red blood cells. After 45 days of storage, the survival rate of red blood cells is still ≥92%, and the hemolysis rate is ≤0.05%, meeting the long-term clinical blood use requirements.

[0020] 2. The present invention abandons the traditional plasticizer DEHP and uses a quaternary ammonium salt-modified stabilizer to simultaneously achieve anticoagulant and antibacterial functions. The introduction of cetyltrimethylammonium bromide significantly increases the killing rate of Staphylococcus aureus on the surface of the blood bag, eliminating the need for additional antibacterial agents and avoiding potential contamination of blood by substances such as silver ions. Meanwhile, the migration rate of the polyolefin base material is less than 0.01 mg / L, far lower than the safety threshold of 0.5 mg / L for DEHP, ensuring blood transfusion safety.

[0021] 3. The present invention forms a C-O-C bonding structure on the polyolefin surface through plasma grafting technology, reducing the water contact angle of the blood bag from 90° to below 55°, reducing the platelet adhesion rate from 15% to 3%, and reducing the complement activation rate from 12% to 4%. The binding strength of the stabilizer coating is ≥5 N / cm, which is 2.5 times that of the coating in the prior art patent CN104403122A, with no risk of peeling during long-term storage, and the hemolysis rate is only 0.05%, generally higher than 0.2% in the prior art.

[0022] 4. The present invention uses a twin-screw co-blending extrusion process to uniformly disperse the stabilizer in the polyolefin material, eliminating the traditional layer-by-layer self-assembly or multi-layer co-extrusion steps, and increasing the production efficiency by 60%. Meanwhile, the synthesis process of the stabilizer does not require noble metal catalysts, reducing the raw material cost by 30%, making it suitable for large-scale industrial production. At the same time, the heat sealing strength of the bag-making is ≥15 N / 15 mm, meeting the GB14232-2004 standard. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a flow chart of a method for preparing a blood stabilizer blood bag proposed by the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0024] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments.

[0025] A blood stabilizer, wherein the stabilizer is a polymer monomer or its oligomer, and the polymer monomer is composed of ethylene glycol, propylene glycol, and glycerol in a molar ratio of 1:1:0.5 - 2; the molecular weight of the stabilizer is 100 - 5000 Da, and a three-dimensional network structure is formed through intermolecular hydrogen bonds.

[0026] The stabilizer is prepared by free radical polymerization reaction. The reaction conditions are as follows: ammonium persulfate is used as an initiator, and the ammonium persulfate accounts for 1-3% of the total mass of the polymerization monomers. The reaction is carried out at 60-80 °C for 2-6 hours; ethylene glycol diglycidyl ether is added as a crosslinking agent during the reaction process to control the molecular weight distribution, and the ethylene glycol diglycidyl ether accounts for 0.5-2% of the total mass of the monomers.

[0027] The stabilizer contains a quaternary ammonium salt modifier, and the quaternary ammonium salt modifier is cetyltrimethylammonium bromide, and the addition amount is 1-2% of the total mass of the polymerization monomers.

[0028] A blood stabilizer blood bag, the blood bag is made of a modified polyolefin material, and the surface of the modified polyolefin material is grafted with the blood stabilizer according to any one of claims 1-3 to form a coating with a thickness of 0.1-5 μm; the water contact angle of the blood bag is ≤60°, the platelet adhesion rate is ≤3%, and the hemolysis rate is ≤0.05%.

[0029] The modified polyolefin material is a polyethylene-polypropylene blend, which is prepared by the following steps:

[0030] a. Under an argon atmosphere, the surface of the film is treated by a plasma processor (power 100-300 W) for 1-3 hours;

[0031] b. Graft the blood stabilizer according to any one of claims 1-3, the treatment temperature is 30-60 °C, and the treatment time is 2-4 hours;

[0032] c. After treatment, a C-O-C bonding structure is formed on the film surface, and the bonding strength is ≥5 N / cm.

[0033] A method for preparing a blood stabilizer blood bag includes the following preparation steps:

[0034] S1. Raw material mixing, mixing the blood stabilizer according to any one of claims 1-3 and the modified polyolefin raw material in a mass ratio of 1:100-1:500;

[0035] S2. Melt blending, melt blending by a twin-screw extruder under the conditions of a temperature of 180-200 °C and a rotation speed of 50-80 revolutions per minute;

[0036] S3. Film blowing and bag making, blowing the melt-blended raw materials into a film with a thickness of 50-100 μm, using the film for blood bag making, and the heat sealing strength after bag making is ≥15 N / 15 mm.

[0037] Example 1

[0038] Performance comparison of blood stabilizers

[0039] Select the blood stabilizer proposed by the present invention, and its specific ratio is ethylene glycol: propylene glycol: glycerol = 1:1:1, and it contains 1.5% cetyltrimethylammonium bromide, denoted as the experimental group;

[0040] Select the PLGA coating material blood stabilizer disclosed in the patent with the publication number "CN104403122A" mentioned in the background technology, denoted as control group 1;

[0041] Select the commercially available propylene glycol blood stabilizer, denoted as control group 2.

[0042] Detect the blood stabilizers of the experimental group, control group 1, and control group 2 respectively. The detection items include: critical micelle concentration, surface tension, antibacterial rate, hemolysis rate, and molecular weight. Record each item of data after detection and make statistics. The specific results are shown in the following table.

[0043] Table 1: Record table of performance comparison data of blood stabilizers

[0044]

[0045] In this embodiment, the critical micelle concentration (CMC): for the experimental group is 0.08 mmol / L, which is reduced by 88% and 81% compared with control group 1 (0.65 mmol / L) and control group 2 (0.42 mmol / L) respectively. The low CMC value indicates that the stabilizer is more likely to form micelles in the blood, effectively reducing the interfacial tension to 28.5 mN / m (control group 1 = 35.2 mN / m, control group 2 = 32.1 mN / m), thereby enhancing the stability of the red blood cell membrane. Molecular weight distribution: the number average molecular weight (Mn) of the experimental group reaches 2800 Da and PDI = 1.3, forming a three-dimensional hydrogen bond network structure (confirmed by strong hydroxyl absorption peaks in FTIR), which has stronger long-term effectiveness and anti-degradation ability compared with the small molecule structures of control group 1 and 2 (PDI = 1.0). According to these two items of data, it can be known that the three-dimensional hydrogen bond network structure significantly improves the performance of the stabilizer.

[0046] Antibacterial rate: the killing rate of the experimental group against Staphylococcus aureus reaches 99.9% (ASTM E2149 standard), while control group 1 and 2 have no antibacterial function. The introduction of cetyltrimethylammonium bromide destroys the bacterial cell membrane through electrostatic interaction and at the same time inhibits platelet aggregation (the hemolysis rate is only 0.02%, control group 1 = 0.15%, control group 2 = 0.21%). Hemolysis rate comparison: the hemolysis rate of the experimental group is reduced by 87% compared with control group 1 and 90% compared with control group 2, indicating that the quaternary ammonium salt modification does not damage the integrity of the red blood cell membrane, but reduces mechanical damage through interfacial stabilization. These two items of data can illustrate that the quaternary ammonium salt modification simultaneously realizes the anticoagulant and antibacterial functions.

[0047] In summary, for the present invention, the specific proportion polymerization of ethylene glycol, propylene glycol, and glycerol is the key to reducing the CMC value. The hydrogen bond network structure enhances the dynamic interaction between the blood stabilizer and the erythrocyte membrane. The introduction of the quaternary ammonium salt modifier breaks through the limitation of the single anticoagulant function of traditional blood stabilizers, achieving a synergistic improvement in antibacterial and blood compatibility.

[0048] Example 2

[0049] Performance comparison of blood bag materials

[0050] Select the modified polyolefin blood bag proposed by the present invention and graft it with the blood stabilizer proposed by the present invention. The specific ratio is ethylene glycol: propylene glycol: glycerol = 1:1:1, and it contains 1.5% cetyltrimethylammonium bromide, denoted as the experimental group;

[0051] Select the PLGA-coated PVC blood bag disclosed in the patent with the publication number "CN104403122A" mentioned in the background technology, denoted as control group 1;

[0052] Select a commercially available DEHP-plasticized PVC blood bag, denoted as control group 2.

[0053] Detect the blood bags of the experimental group, control group 1, and control group 2 respectively. The detection items include: water contact angle, platelet adhesion rate, coating bonding strength, DEHP migration amount, and heat sealing strength. Record each data after detection and make statistics. The specific results are as follows in the table.

[0054] Table 2: Record table of blood bag material performance comparison data

[0055]

[0056] In this example, the water contact angle: for the experimental group is 52°, which is reduced by 33% and 43% compared with control group 1 (78°) and control group 2 (92°) respectively, indicating a significant improvement in the hydrophilicity of the material surface. This is because plasma treatment introduces hydroxyl (-OH) and carbonyl (C=O) groups on the polyolefin surface (confirmed by XPS analysis), enhancing the interfacial affinity with blood. The platelet adhesion rate: for the experimental group is only 2.8%, which is reduced by 71% and 83% compared with control group 1 (9.7%) and control group 2 (16.2%) respectively. The hydrophilic surface effectively inhibits the non-specific binding of platelet membrane glycoproteins (such as GPIIb / IIIa) to the material, reducing the risk of blood coagulation. According to these two data, it can be known that the plasma grafting technology significantly improves the blood compatibility of the material.

[0057] Coating bonding strength: In the experimental group, it reached 5.2 N / cm, which was 2.5 times that of Control Group 1 (2.1 N / cm). The C-O-C bonds formed by plasma grafting (the proportion of the C-O peak area detected by XPS was 18.7%) significantly enhanced the chemical bonding between the coating and the substrate, solving the problem of easy shedding of traditional PLGA coatings (the weight loss rate of Control Group 1 after the accelerated aging test was 12.5%). Long-term stability: After storing at 4°C for 45 days, the coating thickness of the experimental group still remained 98.3% of the initial value (detected by SEM), while the weight loss rate of the coating in Control Group 1 reached 8.2%, verifying the long-term effectiveness of chemical bonding.

[0058] In terms of toxicity control, the migration amount of DEHP in the experimental group was not detected (the detection limit was 0.01 mg / L), basically completely eliminating the carcinogenic risk of traditional PVC blood bags, while the migration amount of DEHP in Control Group 2 was 0.45 mg / L, which was 4.5 times higher than the EU medical device standard (0.1 mg / L). In terms of improving antibacterial properties, the introduction of the quaternary ammonium salt modifier made the killing rate of Escherichia coli on the surface of the experimental group reach 99.8% (ASTM E2149 standard), while Control Groups 1 and 2 had no antibacterial function and needed to add antibacterial agents such as silver ions additionally.

[0059] Heat sealing strength: In the experimental group, it reached 18.5 N / 15 mm, higher than that of Control Group 1 (14.2 N / 15 mm) and Control Group 2 (16.1 N / 15 mm), meeting the GB14232-2004 standard (≥15 N / 15 mm). The twin-screw blending process evenly dispersed the stabilizer (SEM showed that the dispersion coefficient ≤5%), avoiding the decline in mechanical properties caused by too high local concentration.

[0060] In summary, for the present invention, the plasma grafting technology constructs a nano-scale hydrophilic network on the polyolefin surface by precisely controlling the processing parameters, breaking through the short-term effectiveness of traditional physical coating, and combining the quaternary ammonium salt antibacterial group with the stabilizer coating to achieve the trinity function of anticoagulation, antibacterial, and anti-pollution, solving the potential safety hazard of adding antibacterial agents to existing materials. At the same time, the co-extrusion process is compatible with existing bag-making equipment, and the coating bonding strength reaches 5.2 N / cm, which is suitable for large-scale production.

[0061] Example 3

[0062] Comparison of blood preservation effects

[0063] Select the combination of the blood bag + stabilizer proposed in the present invention (the experimental group of Example 1 + the experimental group of Example 2), denoted as the experimental group;

[0064] Select the combination of the blood bag + anticoagulant disclosed in the patent with the publication number "CN104403122A" mentioned in the background technology, denoted as Control Group 1;

[0065] Select a commercially available DEHP plasticized PVC blood bag + anticoagulant combination, denoted as Control Group 2.

[0066] Observe and detect the blood stored in the experimental group, Control Group 1, and Control Group 2 respectively. The items of observation and detection include: blood storage time, red blood cell survival rate, 2,3-DPG retention rate, hemolysis rate, and complement activation rate. After detecting and statistically analyzing each data item, the specific results are shown in the following table.

[0067] Table 3: Record Table of Comparative Data on Blood Preservation Effect

[0068]

[0069] In this embodiment, for the storage time: it reaches 45 days in the experimental group, which is extended by more than 3 days compared with Control Group 1 (42 days), and is extended by 29% compared with Control Group 2 (35 days). This benefits from the inhibition of the glycolysis pathway by the three-dimensional hydrogen bond network stabilizer (ATP content is 1.8 μmol / gHb, Control Group 2 = 1.2 μmol / gHb), and the hydrophilic protective film on the surface of the blood bag material (water contact angle is 52°) reduces the mechanical damage of red blood cells. For the 2,3-DPG retention rate: it is 86.2% in the experimental group, which is increased by 18% compared with Control Group 1 (72.8%), and is increased by 26% compared with Control Group 2 (68.5%). The high retention rate ensures that red blood cells can still effectively release oxygen after transfusion, meeting the clinical emergency blood use requirements.

[0070] For the red blood cell survival rate: it is 92.5% in the experimental group, which is increased by 8.4% compared with Control Group 1 (85.3%), and is increased by 12.7% compared with Control Group 2 (82.1%). Scanning electron microscopy shows that after 45 days of storage, red blood cells still maintain the biconcave disc shape, while obvious shrinkage and membrane damage appear in Control Group 2. For the control of hemolysis rate: it is 0.045% in the experimental group, which is only 19.6% of Control Group 1 (0.23%) and 14.5% of Control Group 2 (0.31%). The quaternary ammonium salt modifier stabilizes the red blood cell membrane through electrostatic action, reducing complement activation (complement activation rate is 4.1%, Control Group 2 = 14.3%).

[0071] In summary, for the present invention, the stabilizer coating and the blood bag substrate are combined through C-O-C bonds (bonding strength is 5.2 N / cm), forming a "protective film - material" dual-stable system, which is more durable than the physical adsorption coating of Control Group 1 (bonding strength is 2.1 N / cm). At the same time, the stabilizer reduces the glycolysis rate by 35% by inhibiting the activity of glucose-6-phosphate dehydrogenase, delaying the consumption of ATP (the ATP content is still 60% of the initial value after 45 days of storage).

[0072] Therefore, through the collaborative innovation of stabilizer molecular design, material interface modification, and process optimization, the present invention has broken through multiple technical bottlenecks in blood preservation time, compatibility, and safety, providing a more efficient and safe solution for clinical blood use.

[0073] The above is only the preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, making equivalent substitutions or changes should be covered within the protection scope of the present invention.

Claims

1. A blood stabilizer, characterized in that: The stabilizer is a polymerized monomer or an oligomer thereof, and the polymerized monomer is composed of ethylene glycol, propylene glycol and glycerol in a molar ratio of 1:1:0.5-2; the molecular weight of the stabilizer is 100-5000Da, and a three-dimensional network structure is formed through intermolecular hydrogen bonds.

2. A blood stabilizer according to claim 1, characterized in that: The stabilizer is prepared by free radical polymerization, and the reaction conditions are: using ammonium persulfate as an initiator, the ammonium persulfate accounts for 1-3% of the total mass of the polymerization monomer, and reacting at 60-80° C. for 2-6 hours; during the reaction, ethylene glycol diglycidyl ether is added as a cross-linking agent to adjust the molecular weight distribution, and the ethylene glycol diglycidyl ether accounts for 0.5-2% of the total mass of the monomer.

3. A blood stabilizer according to claim 1, characterized in that: The stabilizer contains a quaternary ammonium salt modifier, which is hexadecyltrimethylammonium bromide, and the added amount is 1-2% of the total mass of the polymerized monomers.

4. A blood stabilizer blood bag, characterized in that: The blood bag is made of a modified polyolefin material, and the surface of the modified polyolefin material is grafted with the blood stabilizer according to any one of claims 1 to 3 to form a coating with a thickness of 0.1 to 5 μm; The water contact angle of the blood bag is ≤60°, the platelet adhesion rate is ≤3%, and the hemolysis rate is ≤0.05%.

5. A blood stabilizer blood bag according to claim 4, characterized in that: The modified polyolefin material is a polyethylene-polypropylene blend, which is prepared by the following steps: a. In an argon atmosphere, use a plasma treatment machine (power 100~300W) to treat the film surface for 1~3 hours; b. grafting the blood stabilizer according to any one of claims 1 to 3, the treatment temperature is 30 to 60° C., and the treatment time is 2 to 4 hours; c. After treatment, a COC bonding structure is formed on the surface of the film, and the bonding strength is ≥5N / cm.

6. A method for preparing a blood stabilizer blood bag according to any one of claims 1 to 5, characterized in that: The method comprises the following preparation steps: S1. Mixing raw materials: blending the blood stabilizer according to any one of claims 1 to 3 with a modified polyolefin raw material at a mass ratio of 1:100 to 1:500; S2, melt blending, melt blending by a twin-screw extruder at a temperature of 180-200° C. and a rotation speed of 50-80 rpm; S3, film blowing and bag making, the melt-blended raw materials are blown into a film with a thickness of 50-100 μm, and the film is used to make blood bags. The heat sealing strength after bag making is ≥15N / 15mm.

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