WO2021135682A1 - 一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法 - Google Patents

一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法 Download PDF

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WO2021135682A1
WO2021135682A1 PCT/CN2020/128750 CN2020128750W WO2021135682A1 WO 2021135682 A1 WO2021135682 A1 WO 2021135682A1 CN 2020128750 W CN2020128750 W CN 2020128750W WO 2021135682 A1 WO2021135682 A1 WO 2021135682A1
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oxygen
polydopamine
solution
dispersion system
microbubbles
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陈东
孙竹
吴柏衡
盛剑鹏
梁廷波
翟小威
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Zhejiang University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0026—Blood substitute; Oxygen transporting formulations; Plasma extender
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5161—Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5192—Processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

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  • the invention relates to the field of nanotechnology, in particular to a polydopamine nanoparticle stable microbubble dispersion system for intravenous high-efficiency oxygen supply and a preparation method thereof.
  • Intravenous oxygen is an effective method to quickly increase the oxygen content in the blood, which can make the patient quickly out of danger.
  • oxygen cannot be injected directly into the vein.
  • the researchers prepared the microbubble dispersion by high-speed shearing of an aqueous solution containing surfactant molecules, and then injected the microbubble dispersion into the blood by intravenous injection in vitro to achieve the effect of rapid oxygen supply.
  • the dispersion system of microbubbles stabilized by surfactant molecules is unstable and prone to coalescence of microbubbles. Under the action of the Ostwald Ripening effect, gas will gradually transfer from microbubbles with a smaller diameter to microbubbles with a larger diameter. , Which eventually causes the diameter of microbubbles to become larger and larger. Therefore, it is still an urgent problem to prepare a biocompatible, stable and dispersed microbubble system to achieve high-efficiency intravenous oxygen supply.
  • the invention uses biocompatible dopamine as a material to prepare a polydopamine nanoparticle stable microbubble dispersion system for intravenous high-efficiency oxygen supply.
  • the microbubble dispersion system has good dispersibility and stability in water, can realize rapid and efficient delivery of oxygen, and has important application value in the field of intravenous high-efficiency oxygen supply.
  • the present invention discloses a stable microbubble dispersion system of polydopamine nanoparticles for intravenous high-efficiency oxygen supply and a preparation method thereof.
  • dopamine is oxidized by oxygen, and the polydopamine nanoparticles are attached to the interface of the oxygen microbubbles during the polymerization process to form a dense polydopamine particle shell layer.
  • the prepared polydopamine nanoparticle stable oxygen microbubble system has excellent biocompatibility, can realize rapid and efficient oxygen delivery, and has important application value in the field of intravenous efficient oxygen supply.
  • the present invention provides the following solutions:
  • the stable microbubble dispersion system of polydopamine nanoparticles for intravenous high-efficiency oxygen supply of the present invention and the preparation method thereof include the following steps:
  • Oxygen is introduced into the solution obtained in step (1), and the oxygen is cut into microbubbles with a high-speed dispersing homogenizer. Dopamine oxidizes and self-polymerizes under alkaline conditions to form polydopamine nanoparticles, which are attached to the interface of microbubbles to form a dense shell of polydopamine particles. Then turn off the high-speed dispersing homogenizer and continue to feed oxygen until the solution turns brown and black;
  • step (3) Add the glutaraldehyde solution to the solution obtained in step (2), mix it thoroughly with a high-speed dispersing homogenizer, and then stir to solidify the polydopamine particle shell layer attached to the microbubble interface;
  • step (3) Dilute the polydopamine nanoparticles stabilized microbubble dispersion solution obtained in step (3), and remove excess polydopamine nanoparticles and other reagents that are not adsorbed on the microbubble interface through filtration and washing, and finally obtain stable water A stable oxygen microbubble dispersion system of dispersed polydopamine nanoparticles.
  • the concentration of dopamine is 10-40 mg/mL, the concentration of polylysine is 1-40 mg/ml, and the concentration of chitosan quaternary ammonium salt is 20-400 mg/ml;
  • the mass ratio of the polylysine to the dopamine hydrochloride is 0.25-1; the mass ratio of the chitosan to the dopamine hydrochloride is 2-10.
  • adding the tris buffer solution to adjust the pH of the mixed solution to make the mixed solution alkaline is to add Tris-HCl to the mixed solution to adjust the pH of the mixed solution to 7.5-9.0.
  • the flow rate of oxygen gas is 0.5-2L/min
  • the rotation speed of the high-speed dispersing homogenizer is 10000rpm-14000rpm
  • the homogenization time is 3-6min; the oxygen gas is continuously fed until the solution becomes The brown-black time is 1-5min.
  • the volume fraction of the glutaraldehyde solution is 2%-6.5%, the amount of the glutaraldehyde solution is 1ml-5ml; the rotation speed of the high-speed dispersing homogenizer is 10000rpm-14000rpm, and the homogenization
  • the time is 1-4 min; the stirring is performed at 20°C-40°C, and the stirring time is 30 min-2h.
  • the pore size of the filter paper is 1um-11um.
  • the invention also discloses a stable oxygen microbubble dispersion system of polydopamine particles prepared by the method.
  • the microbubble dispersion system has excellent biocompatibility, can stably carry oxygen for a long time, and can be used in extremely hypoxic solutions. It can release oxygen quickly and efficiently, supply oxygen to hypoxic tissue, and has important application value in the field of high-efficiency intravenous oxygen supply.
  • the present invention proposes a polydopamine nanoparticle stable microbubble dispersion system for intravenous high-efficiency oxygen supply and a preparation method thereof. This method solves the problem of low oxygen carrying capacity in the traditional surfactant molecule-stabilized microbubble dispersion system , The system is unstable, which makes it difficult to achieve intravenous oxygen supply and other problems.
  • the method used in the present invention does not produce harmful substances during the experiment.
  • the prepared polydopamine nanoparticles and stable oxygen microbubbles have good biocompatibility, are non-toxic and non-irritating to the human body, and can be applied to biological organisms. Medical field.
  • the interface of the microbubbles prepared by the present invention is covered with a relatively dense polydopamine nanoparticle shell layer, which can well prevent the fusion of the bubbles, so that the microbubbles can stably carry oxygen for a long time.
  • the stabilized oxygen microbubbles of the polydopamine nanoparticles prepared by the present invention can effectively control the rapid release of oxygen contained in them in an extremely hypoxic solution, and provide a basis for realizing high-efficiency intravenous oxygen supply.
  • Figure 1 is a schematic diagram of an apparatus for preparing a stable microbubble dispersion system of polydopamine nanoparticles
  • Example 2 is a schematic diagram of the attachment and crosslinking scheme of polydopamine nanoparticles at the oxygen-water interface in Example 1 of the present invention
  • FIG. 3a is an optical image of the stabilized microbubble dispersion solution of polydopamine nanoparticles obtained in step (3) in Example 1 of the present invention
  • FIG. 3b is an optical image of the stable oxygen microbubble dispersion of polydopamine nanoparticles obtained in step (4) in Example 1 of the present invention
  • FIG. 4 is the electric potential measurement of the polydopamine nanoparticles stabilized microbubble dispersion in Example 1 of the present invention
  • FIG. 5 is an optical image of the stable microbubble morphology of polydopamine nanoparticles changing with time in Example 2 of the present invention
  • Figure 6a is a scanning electron microscope image of polydopamine nanoparticles stabilized microbubbles in Example 2 of the present invention
  • Fig. 6b is a scanning electron microscope image when the stabilized microbubbles of polydopamine nanoparticles collapsed in Example 2 of the present invention
  • FIG. 7 is a time-intensity curve of the release of oxygen in vitro by the stabilized microbubbles of polydopamine nanoparticles in Example 3 of the present invention.
  • Example 1 Preparation of stable oxygen microbubble dispersion system of polydopamine nanoparticles
  • the method of the present invention is used to prepare a polydopamine nanoparticle stable oxygen microbubble dispersion system.
  • the specific steps are as follows:
  • step (2) Feed oxygen into the solution obtained in step (1) at a flow rate of 1 L/min, adjust the speed of the high-speed dispersion homogenizer to 12000 rpm, and shear the oxygen into microbubbles with the homogenizer.
  • Dopamine oxidizes and self-polymerizes under alkaline conditions to form polydopamine nanoparticles, which adhere to the interface of microbubbles to form a dense shell of polydopamine particles. (As shown in Figure 2), after homogenizing for 5 minutes, turn off the high-speed dispersing homogenizer, and continue to inject oxygen for 2 minutes until the solution is brown-black.
  • step (3) Add 1ml of 4% glutaraldehyde solution to the solution obtained in step (2), and homogenize with a high-speed dispersing homogenizer at 12000rpm for 3min, and then stir at 1000rpm for 30min at room temperature to solidify the adhesive attached to the microbubble interface
  • the polydopamine particle shell layer is used to obtain a stable microbubble dispersion solution of polydopamine nanoparticles, as shown in Figure 3a.
  • step (3) Dilute the polydopamine nanoparticle stable microbubble dispersion solution obtained in step (3) with deionized water 5 times, and then filter the diluted solution with a filter paper with a pore size of 2.5um and a glass funnel to remove excess
  • the polydopamine nanoparticles and other reagents that are not adsorbed on the interface of the microbubbles are then washed with deionized water and filtered again with the microbubbles remaining on the filter paper.
  • the microbubbles are washed with deionized water and stored in a glass bottle to obtain a A stable dispersion of oxygen microbubbles of polydopamine nanoparticles dispersed in water, as shown in Figure 3b.
  • the surface of the microbubbles has a positive charge of +62.5mV, which proves that there is a polydopamine shell on the surface of the microbubbles.
  • Example 2 Changes in the stable microbubble morphology of polydopamine nanoparticles over time.
  • the dispersion of the stable microbubbles of polydopamine nanoparticles in water was stored at room temperature. Observe the changes of the microbubble morphology in the water regularly through an optical microscope, as shown in Figure 5.
  • the surface morphology of the stable microbubbles of the polydopamine particles was observed through a scanning electron microscope. As shown in Figure 6a, there are dense nano-sized polydopamine particles at the interface of the microbubbles. After being stored for one week, some of the stable microbubbles of the polydopamine nanoparticles shrank or even collapsed, resulting in the release of oxygen wrapped in the shell of the polydopamine nanoparticles, as shown in Figure 6b.
  • Example 3 Oxygen release rate in vitro from stabilized oxygen microbubbles of polydopamine nanoparticles

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Abstract

一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法。该方法将多巴胺、壳聚糖季铵盐和富氨基聚合物溶于水中，调节其pH至碱性，然后向溶液中通入氧气。在均质机的强剪切力作用下，氧气氧化多巴胺，得到的聚多巴胺纳米颗粒在聚合过程中附着在氧气微气泡的界面，形成一层致密的聚多巴胺颗粒壳层。最后加入戊二醛固化附着在微气泡界面的聚多巴胺颗粒壳层，并通过过滤、洗涤和再分散，得到在水中稳定分散的氧气微气泡。该纳米颗粒稳定的氧气微气泡，具有优异的生物相容性，可实现氧气的快速高效递送，在静脉高效给氧领域具有重要应用价值。

Description

一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法 技术领域
本发明涉及纳米技术领域，具体涉及一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法。
背景技术
静脉高效给氧一直是人们面临的一个重要挑战。窒息性心脏骤停常出现于患有严重或长期缺氧的病人，当血液中的氧气过低时，心脏停止跳动，氧气停止输送到生命器官。如果不能迅速恢复血氧水平，几分钟内就会导致器官损伤和死亡。目前抢救窒息性心脏骤停患者主要通过吸氧、插管或机械通气等治疗手段。然而，当这些治疗手段无法快速提高患者血液中氧气含量时，可能直接威胁到患者生命健康，导致该病死亡率较高。
静脉给氧是一种快速提高血液中氧气含量的有效方法，可使患者快速脱离危险。但由于氧气在血液中溶解度低，无法直接将氧气注射到静脉中。研究者通过高速剪切含有表面活性剂分子的水溶液，制备得到微气泡分散液，然后通过体外静脉注射将微气泡分散液注射到血液中，以达到快速给氧的效果。然而由表面活性剂分子稳定的微气泡分散体系不稳定，易发生微气泡的聚并，并且在Ostwald Ripening效应的作用下，气体会逐渐从直径较小的微气泡往直径较大的微气泡转移，最终导致微气泡的直径越来越大。因此制备生物相容、稳定分散的微气泡体系，实现静脉高效给氧仍是一项亟待解决的问题。
本发明采用生物相容的多巴胺为材料，制备了一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系。该微气泡分散体系在水中具有良好的分散性和稳定性，可实现氧气的快速高效递送，在静脉高效给氧领域具有重要应用价值。
发明内容
针对现有技术的不足，本发明公开了一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法。该方法通过氧气氧化多巴胺，并在聚合过程中使聚多巴胺纳米颗粒附着在氧气微气泡的界面，形成一层致密的聚多巴胺颗粒壳层。制备的聚多巴胺纳米颗粒稳定的氧气微气泡体系，具有优异的生物相容性，可实现氧气的快速高效递送，在静脉高效给氧领域具有重要应用价值。
为实现上述目的，本发明提供如下解决方案：
本发明静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法包括以下步骤：
(1)将多巴胺、壳聚糖季铵盐和聚赖氨酸溶于去离子水中，然后加入tris缓冲溶液调节混合溶液pH值，使该混合溶液呈碱性；
(2)向步骤(1)所得溶液中通入氧气，并用高速分散均质机将氧气剪切成微气泡。多巴胺在碱性条件下氧化自聚形成聚多巴胺纳米颗粒，并附着在微气泡界面，形成一层致密的聚多巴胺颗粒壳层。随后关闭高速分散均质机，继续通入氧气至溶液呈棕黑色；
(3)向步骤(2)所得溶液中加入戊二醛溶液，并用高速分散均质机充分混合，然后搅拌，使其固化附着在微气泡界面的聚多巴胺颗粒壳层；
(4)将步骤(3)中所得聚多巴胺纳米颗粒稳定的微气泡分散溶液稀释，并通过过滤、洗涤去除多余的未吸附在微气泡界面的聚多巴胺纳米颗粒和其他试剂，最后得到在水中稳定分散的聚多巴胺纳米颗粒稳定的氧气微气泡分散体系。
优选的，所述步骤(1)中，多巴胺的浓度为10-40mg/mL，聚赖氨酸的浓度为1-40mg/ml，壳聚糖季铵盐的浓度为20-400mg/ml；所述聚赖氨酸与所述盐酸多巴胺的质量比为0.25-1；所述壳聚糖与所述盐酸多巴胺的质量比为2-10。
优选的，所述步骤(1)中，所述加入tris缓冲溶液调节混合溶液pH值，使该混合溶液为碱性，是向该混合溶液中加入Tris-HCl使该混合溶液的pH值调节至7.5-9.0。
优选的，所述步骤(2)中，通入氧气的流速为0.5-2L/min，高速分散均质机的转速为10000rpm-14000rpm，均质时间为3-6min；继续通入氧气至溶液呈棕黑色的时间为1-5min。
优选的，所述步骤(3)中，戊二醛溶液的体积分数为2％-6.5％，戊二醛溶液的用量为1ml-5ml；高速分散均质机的转速为10000rpm-14000rpm，均质时间为1-4min；所述搅拌是在20℃-40℃下进行的搅拌时间为30min-2h。
优选的，所述步骤(4)中，滤纸的孔径为1um-11um。
本发明还公开了所述方法制备得到的聚多巴胺颗粒稳定的氧气微气泡分散体系，所述的微气泡分散体系具有优异的生物相容性，可长时间稳定载氧，在极度缺氧溶液中可实现快速、高效地释放氧气，为缺氧组织供氧，在静脉高效给氧领域具有重要应用价值。
本发明的有益效果如下：
(1)本发明提出一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系及其制备方法，该方法解决了传统表面活性剂分子稳定的微气泡分散体系存在载氧量低、体系不稳定、从而难以实现静脉给氧等问题。
(2)本发明所用方法，实验过程中不会有有害物质产生，所制得的聚多巴胺纳米颗粒稳定的氧气微气泡具有良好的生物相容性，对人体无毒无刺激，可应用于生物医学领域。
(3)本发明所制得的微气泡界面覆盖着相对致密的聚多巴胺纳米颗粒壳层，能很好地防止气泡融合，使微气泡可以长时间稳定载氧。
(4)本发明所制得的聚多巴胺纳米颗粒稳定的氧气微气泡可以在极度缺氧溶液中有效地控制包裹在其内的氧气的快速释放，为实现静脉高效给氧提供基础。
附图说明
图1为制备聚多巴胺纳米颗粒稳定的微气泡分散体系的装置示意图；
图2为本发明实施例1中聚多巴胺纳米颗粒在氧气-水界面附着交联的方案示意图；
图3a为本发明实施例1中步骤(3)中得到的聚多巴胺纳米颗粒稳定的微气泡分散溶液的光学图像；
图3b为本发明实施例1中步骤(4)中得到的聚多巴胺纳米颗粒稳定的氧气微气泡分散液的光学图像；
图4为本发明实施例1中聚多巴胺纳米颗粒稳定的微气泡分散液的电位测定；
图5为本发明实施例2中聚多巴胺纳米颗粒稳定的微气泡形态随时间变化的光学图像；
图6a为本发明实施例2中聚多巴胺纳米颗粒稳定的微气泡的扫描电镜图；
图6b为本发明实施例2中聚多巴胺纳米颗粒稳定的微气泡发生坍塌时的扫描电镜图；
图7为本发明实施例3中聚多巴胺纳米颗粒稳定的微气泡在体外释放氧气的时间-强度曲线。
具体实施方式
下面举实施例说明本发明，但本发明并不限于下述的实施例。
实施例1：制备聚多巴胺纳米颗粒稳定的氧气微气泡分散体系
参照附图1中的装置，采用本发明的方法制备聚多巴胺纳米颗粒稳定的氧气微气泡分散体系，具体步骤如下：
(1)将20mg多巴胺，5mg聚赖氨酸和120mg壳聚糖季铵盐溶于10mL去离子水中，然后加入1mL的pH＝8.5的tris盐缓冲溶液调节混合溶液pH值，使该混合溶液为碱性。
(2)向步骤(1)所得溶液中以1L/min的流速通入氧气，调节高速分散均质机转速为12000rpm，用均质机将氧气剪切成微气泡。多巴胺在碱性条件下氧化自聚形成聚多巴胺纳米颗粒，并附着在微气泡界面，形成一层致密的聚多巴胺颗粒壳层。(如附图2所示)，均质5min后关闭高速分散均质机，继续通入氧气2min至溶液呈棕黑色。
(3)向步骤(2)所得溶液中加入1ml的4％戊二醛溶液，并用高速分散均质机在12000rpm下均质3min，然后在室温下以1000rpm搅拌30min，固化附着在微气泡界面的聚多巴胺颗粒壳层,得到聚多巴胺纳米颗粒稳定的微气泡分散溶液，如附图3a所示。
(4)将步骤(3)中所得聚多巴胺纳米颗粒稳定的微气泡分散溶液用去离子水稀释5倍，然后用孔径为2.5um的滤纸和玻璃漏斗对稀释后的溶液进行过滤，去除多余的未吸附在微气泡界面的聚多巴胺纳米颗粒和其他试剂，然后将留在滤纸上的微气泡用去离子水洗涤，再次过滤，最后用去离子水将微气泡冲入玻璃瓶中储存，得到在水中稳定分散的聚多巴胺纳米颗粒稳定的氧气微气泡分散液,如 附图3b所示。微气泡表面带有+62.5mV的正电荷，证明微气泡表面存在聚多巴胺壳层。
实施例2：聚多巴胺纳米颗粒稳定的微气泡形态随时间的变化。
将聚多巴胺纳米颗粒稳定的微气泡在水中的分散液于室温下静置储存。通过光学显微镜定期观察水中微气泡形态的变化，如附图5所示。通过扫描电子显微镜观察聚多巴胺颗粒稳定的微气泡的表面形态，如附图6a所示，其微气泡界面存在致密的纳米尺寸的聚多巴胺颗粒。静置储存一周后，部分聚多巴胺纳米颗粒稳定的微气泡的形态发生皱缩甚至坍塌，导致包裹在聚多巴胺纳米颗粒壳层中的氧气释放出来，如附图6b所示。
实施例3：聚多巴胺纳米颗粒稳定的氧气微气泡体外释放氧气速率
(1)20℃室温条件下，用纯N 2将生理盐水溶解氧浓度降低为0mg/L，制成极度缺氧溶液。在聚多巴胺纳米颗粒稳定的氧气微气泡制备成功后的6h后，将1ml载氧微泡分散液注入密闭的10ml的极度缺氧溶液中，采用溶解氧测定仪检测溶液中溶解氧变化情况，连续观察约30min载氧微泡在极度缺氧生理盐水中释放O 2的情况，如附图7所示。聚多巴胺纳米颗粒稳定的氧气微气泡在极度缺氧生理盐水中能够有效释放O 2，1ml载氧微泡分散液在800s内使溶液溶解氧量增加至～5.5mg/ml，基本恢复到水中的饱和氧气含量范围内。
(2)20℃室温条件下，用纯N 2将生理盐水溶解氧浓度降低为0mg/L，制成极度缺氧溶液。在聚多巴胺纳米颗粒稳定的氧气微气泡制备成功后的6h后，将2ml载氧微泡分散液注入密闭的10ml的极度缺氧溶液中，采用溶解氧测定仪检测溶液中溶解氧变化情况，连续观察约30min载氧微泡在极度缺氧生理盐水中释放O 2的情况，如附图7所示。聚多巴胺纳米颗粒稳定的氧气微气泡在极度缺氧生理盐水中能够有效释放O 2，2ml载氧微泡分散液在500s内使溶液溶解氧量增加至～5.5mg/ml，基本恢复到水中的饱和氧气含量范围内。
(3)20℃室温条件下，用纯N 2将生理盐水溶解氧浓度降低为0mg/L，制成极度缺氧溶液。在聚多巴胺纳米颗粒稳定的氧气微气泡制备成功后的6h后，将3ml载氧微泡分散液注入密闭的10ml的极度缺氧溶液中，采用溶解氧测定仪检测溶液中溶解氧变化情况，连续观察约30min载氧微泡在极度缺氧生理盐水中释放O 2的情况，如附图7所示。聚多巴胺纳米颗粒稳定的氧气微气泡在极度缺氧 生理盐水中能够有效释放O 2，3ml载氧微泡分散液在200s内使溶液溶解氧量增加至～5.5mg/ml，基本恢复到水中的饱和氧气含量范围内。

Claims (5)

  1. 一种用于静脉高效给氧的聚多巴胺纳米颗粒稳定的微气泡分散体系的制备方法，其特征在于，包括以下步骤：
    (1)将多巴胺、壳聚糖季铵盐和聚赖氨酸溶于去离子水中，然后加入tris缓冲溶液调节混合溶液pH值，使该混合溶液呈碱性；其中多巴胺的浓度为10-40mg/mL，聚赖氨酸的浓度为1-40mg/ml，壳聚糖季铵盐的浓度为20-400mg/ml；所述聚赖氨酸与所述多巴胺的质量比为0.25-1；所述壳聚糖与所述多巴胺的质量比为2-10；
    (2)向步骤(1)所得溶液中通入氧气，并用高速分散均质机将氧气剪切成微气泡，多巴胺在碱性条件下氧化自聚形成聚多巴胺纳米颗粒，并附着在微气泡界面，形成一层致密的聚多巴胺颗粒壳层，随后关闭高速分散均质机，继续通入氧气至溶液呈棕黑色；
    (3)向步骤(2)所得溶液中加入戊二醛溶液，并用高速分散均质机充分混合，然后搅拌，使其固化附着在微气泡界面的聚多巴胺颗粒壳层；其中戊二醛溶液的体积分数为2％-6.5％，戊二醛溶液的用量为1ml-5ml；高速分散均质机的转速为10000rpm-14000rpm，均质时间为1-4min；所述搅拌是在20℃-40℃下进行的，搅拌时间为30min-2h；
    (4)将步骤(3)中所得聚多巴胺纳米颗粒稳定的微气泡分散溶液稀释，并通过过滤、洗涤去除多余的未吸附在微气泡界面的聚多巴胺纳米颗粒和试剂，最后得到在水中稳定分散的聚多巴胺纳米颗粒稳定的氧气微气泡分散体系。
  2. 根据权利要求1所述的聚多巴胺纳米颗粒稳定的微气泡分散体系的制备方法，其特征在于，所述步骤(1)中，所述加入tris缓冲溶液调节混合溶液pH值，使该混合溶液为碱性，是向该混合溶液中加入Tris-HCl使该混合溶液的pH值调节至7.5-9.0。
  3. 根据权利要求1所述的聚多巴胺纳米颗粒稳定的微气泡分散体系的制备方法，其特征在于，所述步骤(2)中，通入氧气的流速为0.5-2L/min，高速分散均质机的转速为10000rpm-14000rpm，均质时间为3-6min；继续通入氧气至溶液呈棕黑色。
  4. 根据权利要求1所述的聚多巴胺纳米颗粒稳定的微气泡分散体系的制备方法，其特征在于，所述步骤(4)中，滤纸的孔径为1um-11um。
  5. 权利要求1-4任一项所述方法制备得到的聚多巴胺纳米颗粒稳定的微气泡分散体系。
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