Platelet membrane self-assembly nanobubble, and preparation method and application thereof
A nano-bubble and platelet technology, which is applied in the field of biomedicine, can solve the problems of difficulty in the targeted ultrasound imaging diagnosis of small and micro lesions, and achieve the effects of good biological safety, good repeatability, and prolonged circulation time.
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Embodiment 1
[0033] Set the concentration to 1 x 109 Fresh platelets per milliliter were separated by centrifugation at 500g for 10min, resuspended and washed with normal saline to remove plasma, and then placed in a -80°C refrigerator to freeze, frozen platelets were thawed at room temperature, and centrifuged at 4000g for 5min to separate, and The saline solution was resuspended and washed three times to separate the platelet membrane vesicles from the organelles to obtain a purified platelet membrane vesicle suspension. After the obtained platelet vesicle suspension was homogenized by 100W, 42KHz water bath ultrasound for 5min, it was broken by ultrasonic cavitation at 500W power for 30s, and sulfur hexafluoride gas was introduced during this process; to reduce ultrasonic cavitation Power to 100W for 90s, so that the platelet membrane fragments broken by ultrasonic cavitation are adsorbed on the nano-nucleus air-liquid interface formed by cavitation, and recombined to form a suspension o...
Embodiment 2
[0036] The platelet membrane extraction method in this example is the same as that in Example 1. The device for preparing nano-bubbles, the fixed volume of the vial is 3mL; the volume of the syringe of the volume-variable extrusion device can be changed in the range of 0-5mL; the volume of the connecting tube is 0.5mL, one end is connected to the volume-variable device, and the other end is inserted into the generating container for the vial Bottles containing water-purified platelet membrane vesicle suspensions.
[0037] 2 mL of platelet membrane vesicle suspension and 4.5 mL of 99.99% high-purity sulfur hexafluoride (SF 6 ) is accommodated in a nanobubble device to form a closed system. At this time, the equilibrium volume of the variable volume extruding device is 3mL; 6 Pressed into the water through the connecting pipe, at this time SF 6 The total volume is 1.5mL, and the system pressure is 0.3MPa; remove the external force, return the column sieve to the equilibrium po...
Embodiment 3
[0040] In this example, the platelet membrane extraction method is the same as in Example 1; the platelet membrane-coated nanobubble preparation method is the same as in Example 2, the only difference is that the sulfur hexafluoride gas used in Example 1 is replaced by helium. The microstructure of the prepared platelet membrane nanobubbles was characterized by a transmission electron microscope, which has a good bubble structure, and the assembly structure of the platelet membrane on the surface of the nanobubbles can be clearly observed.
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