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Experimental Study on the Compatibility and Characteristics of a Dual-Target Microbubble Loaded with Anti-miR-33
International Journal of Nanomedicine ( IF 6.6 ) Pub Date : 2021-09-11 , DOI: 10.2147/ijn.s324514 Chen Yuan 1 , Yanhong Li 1 , Liyun Liu 1 , Baihetiya Tayier 1 , Lingjie Yang 1 , Lina Guan 1, 2, 3 , Yuming Mu 1, 2, 3
International Journal of Nanomedicine ( IF 6.6 ) Pub Date : 2021-09-11 , DOI: 10.2147/ijn.s324514 Chen Yuan 1 , Yanhong Li 1 , Liyun Liu 1 , Baihetiya Tayier 1 , Lingjie Yang 1 , Lina Guan 1, 2, 3 , Yuming Mu 1, 2, 3
Affiliation
Objective: To prepare a new type of dual-target microbubble loaded with anti-miR-33 (ANM33).
Methods: Carrier core nanobubbles (NBs) were prepared by thin film hydration, and microbubbles loaded with PM1 (PCNBs) were prepared by grafting DSPE-PEG2000-maleimide-PM1 onto the NB surface. ANM33 was connected via electrostatic adsorption and covalent bonding, and hyaluronic acid (HA) was covalently connected. PM1 and HA were the targets, and ANM33 was the intervention drug. To evaluate the general physical and chemical properties of the prepared dual-target microbubbles loaded with ANM33 (HA-PANBs), we observed their morphology, particle size and surface potential while monitoring their stability and in vitro imaging ability, evaluated their toxic effect on cells and verified their ability to target cells.
Results: HA-PANBs had a regular morphology and good stability. The average particle size measured by a Malvern potentiometer was 1421.75± 163.23 nm, and the average surface potential was − 5.51± 1.87 mV. PM1 and ANM33 were effectively connected to the NBs. The PM1, ANM33, and HA binding reached 89.0± 1.1%, 65.02± 5.0%, and 61.4± 3.5%, respectively, and the maximum binding reached 2 μg, 5 μg, and 7 μg/108 microbubbles, respectively. HA-PANBs had no obvious toxic effects on cells, and their ability to continuously enhance imaging in vitro persisted for more than 15 minutes, obviously targeting foam cells in the early stage of AS.
Conclusion: HA-PANBs are ideal ultrasound contrast agents. The successful, firm connection of PM1 and HA to the NBs significantly increased the amount of carried ANM33. When microbubbles prepared with 2:4:7 PM1:ANM33:HA were used as a contrast agent, they had a high ANM33 carrying capacity, stable physical properties, and significantly enhanced imaging and targeting of foam cells in the early stage of AS.
Keywords: ultrasound microbubbles, molecular targeting, dual targeting, atherosclerosis
中文翻译：
载有Anti-miR-33的双靶点微泡相容性和特性的实验研究
目的：制备一种新型双靶点抗miR-33（ANM33）微泡。
方法：通过薄膜水化制备载体核纳米气泡（NBs），并通过将DSPE-PEG2000-马来酰亚胺-PM1接枝到NB表面制备负载PM1的微泡（PCNBs）。ANM33通过静电吸附和共价键连接，透明质酸（HA）共价连接。PM1和HA为靶点，ANM33为干预药物。为了评估所制备的载有 ANM33 (HA-PANB) 的双靶点微泡的一般物理和化学性质，我们观察了它们的形态、粒径和表面电位，同时监测了它们的稳定性和体外成像能力，评估了它们对细胞的毒性作用并验证了它们靶向细胞的能力。
结果：HA-PANBs具有规则的形态和良好的稳定性。Malvern电位计测得的平均粒径为1421.75±163.23 nm，平均表面电位为-5.51±1.87 mV。PM1 和 ANM33 有效地连接到 NB。PM1、ANM33和HA的结合分别达到89.0±1.1%、65.02±5.0%和61.4±3.5%，最大结合分别达到2 μg、5 μg和7 μg/10 8个微泡。HA-PANBs对细胞无明显毒性作用，其体外持续增强成像能力持续15分钟以上，明显针对AS早期的泡沫细胞。
结论：HA-PANB 是理想的超声造影剂。PM1 和 HA 与 NB 的成功、牢固连接显着增加了 ANM33 的携带量。以2:4:7 PM1:ANM33:HA制备的微泡作为造影剂时，具有较高的ANM33承载能力、稳定的物理性能，显着增强了AS早期泡沫细胞的成像和靶向性。
关键词：超声微泡，分子靶向，双重靶向，动脉粥样硬化
更新日期：2021-09-12
Methods: Carrier core nanobubbles (NBs) were prepared by thin film hydration, and microbubbles loaded with PM1 (PCNBs) were prepared by grafting DSPE-PEG2000-maleimide-PM1 onto the NB surface. ANM33 was connected via electrostatic adsorption and covalent bonding, and hyaluronic acid (HA) was covalently connected. PM1 and HA were the targets, and ANM33 was the intervention drug. To evaluate the general physical and chemical properties of the prepared dual-target microbubbles loaded with ANM33 (HA-PANBs), we observed their morphology, particle size and surface potential while monitoring their stability and in vitro imaging ability, evaluated their toxic effect on cells and verified their ability to target cells.
Results: HA-PANBs had a regular morphology and good stability. The average particle size measured by a Malvern potentiometer was 1421.75± 163.23 nm, and the average surface potential was − 5.51± 1.87 mV. PM1 and ANM33 were effectively connected to the NBs. The PM1, ANM33, and HA binding reached 89.0± 1.1%, 65.02± 5.0%, and 61.4± 3.5%, respectively, and the maximum binding reached 2 μg, 5 μg, and 7 μg/108 microbubbles, respectively. HA-PANBs had no obvious toxic effects on cells, and their ability to continuously enhance imaging in vitro persisted for more than 15 minutes, obviously targeting foam cells in the early stage of AS.
Conclusion: HA-PANBs are ideal ultrasound contrast agents. The successful, firm connection of PM1 and HA to the NBs significantly increased the amount of carried ANM33. When microbubbles prepared with 2:4:7 PM1:ANM33:HA were used as a contrast agent, they had a high ANM33 carrying capacity, stable physical properties, and significantly enhanced imaging and targeting of foam cells in the early stage of AS.
Keywords: ultrasound microbubbles, molecular targeting, dual targeting, atherosclerosis
中文翻译：
载有Anti-miR-33的双靶点微泡相容性和特性的实验研究
目的：制备一种新型双靶点抗miR-33（ANM33）微泡。
方法：通过薄膜水化制备载体核纳米气泡（NBs），并通过将DSPE-PEG2000-马来酰亚胺-PM1接枝到NB表面制备负载PM1的微泡（PCNBs）。ANM33通过静电吸附和共价键连接，透明质酸（HA）共价连接。PM1和HA为靶点，ANM33为干预药物。为了评估所制备的载有 ANM33 (HA-PANB) 的双靶点微泡的一般物理和化学性质，我们观察了它们的形态、粒径和表面电位，同时监测了它们的稳定性和体外成像能力，评估了它们对细胞的毒性作用并验证了它们靶向细胞的能力。
结果：HA-PANBs具有规则的形态和良好的稳定性。Malvern电位计测得的平均粒径为1421.75±163.23 nm，平均表面电位为-5.51±1.87 mV。PM1 和 ANM33 有效地连接到 NB。PM1、ANM33和HA的结合分别达到89.0±1.1%、65.02±5.0%和61.4±3.5%，最大结合分别达到2 μg、5 μg和7 μg/10 8个微泡。HA-PANBs对细胞无明显毒性作用，其体外持续增强成像能力持续15分钟以上，明显针对AS早期的泡沫细胞。
结论：HA-PANB 是理想的超声造影剂。PM1 和 HA 与 NB 的成功、牢固连接显着增加了 ANM33 的携带量。以2:4:7 PM1:ANM33:HA制备的微泡作为造影剂时，具有较高的ANM33承载能力、稳定的物理性能，显着增强了AS早期泡沫细胞的成像和靶向性。
关键词：超声微泡，分子靶向，双重靶向，动脉粥样硬化
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