| 摘要: |
| 目的 制备一种新型脂质纳米探针(HD@P-NPs),探究体外超声显像效果,观察其对于瘢痕疙瘩成纤维细胞的迁移抑制以及级联放大治疗疗效。方法 本研究以固体脂质纳米复合体为药物载体,核心负载全氟己烷(PFH),同时搭载血卟啉单甲醚(HMME)及阿霉素(DOX),采用超声乳化法制备HD@P-NPs纳米探针,观察纳米探针微观形态并检测其粒径、电位、PDI分散性及稳定性,计算相应药物包封率及载药率,探究HD@P-NPs在LIFU作用下的超声(US)及对比增强超声(CEUS)成像效果；取对数生长的瘢痕疙瘩成纤维细胞KFS与不同实验组共培养,并分为以下五组：对照组、单纯LIFU辐照组、D@P-NPs组、HD@P-NPs组、HD@P-NPs+LIFU辐照组,使用钙黄绿素-AM/碘化丙啶染色法评估纳米探针HD@P-NPs声动力疗效,采用MTT法检测细胞存活率,研究KFS细胞迁移能力并应用DCFH-DA指示剂观察细胞内活性氧产生。结果 ①制备的纳米探针HD@P-NPs尺寸均一、稳定性好,平均粒径为170.36±6.03nm,电位为-36.91±3.56mv,HMME包封率和载药率分别为67.41%和5.18%,DOX包封率和载药率分别为72.80%和11.20%。②LIFU辐照后,脂质纳米探针HD@P-NPs相变产生微气泡,在3W/cm2、2min条件下可实现最大显像效果。③MTT和钙黄绿色-AM/碘化丙啶染色结果显示,纳米探针HD@P-NPs在联合LIFU条件下对KFS细胞具有明显细胞抑制作用,呈明显红色荧光,而单纯LIFU照射未见任何细胞抑制作用,同时D@P-NPs组与HD@P-NPs组细胞抑制作用较HD@P-NPs+LIFU组较弱,且两组间细胞抑制无统计学意义(P＞0.05)。④KFS细胞迁移实验显示,在无LIFU作用下D@P-NPs组与HD@P-NPs组细胞迁移抑制无统计学意义(P＞0.05),而HD@P-NPs+LIFU组显示出最大细胞迁移抑制作用,迁移率仅为10.32%(P＜0.05),并且在LIFU作用下KFS细胞ROS产量明显增加。结论 成功制备了脂质纳米探针HD@P-NPs,在联合LIFU作用下可提高靶区有效药物浓度,实现超声可视化下KFS细胞声动力级联放大治疗。 |
| 关键词: 声动力治疗 脂质纳米探针 瘢痕疙瘩成纤维细胞 血卟啉单甲醚 阿霉素 超声成像 |
| DOI： |
| 投稿时间：2024-04-29修订日期：2024-05-18 |
| 基金项目:泸州市指导性科技计划项目；西南医科大学附属医院博士启动资金项目 |
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| Cascade therapy study of keloid fibroblasts with phase-change nanoprobes |
| FAN Zhengchao,XIA Jizhu,ZHU Weiwei,XU Ying,ZHAO Xiangzhi |
| (Department of Ultrasound,Affiliated Hospital of Southwest Medical University) |
| Abstract: |
| Objective To prepare a new type of lipid nanoprobes (HD@P-NPs), investigate their in vitro ultrasound imaging effects, and observe their migration inhibition of keloid fibroblasts as well as their therapeutic efficacy in cascade amplification. Methods In this study, solid lipid nanocomposites were used as drug carriers, core-loaded with perfluorohexane (PFH), and piggybacked with hematoporphyrin monomethyl ether (HMME) and Doxorubicin(DOX), and ultrasonic emulsification was used to prepare HD@P-NPs nano-probes, to observe the microscopic morphology of the nano-probes and to detect their particle sizes, potentials, PDI dispersion, and stability, and to calculate the encapsulation rate of the corresponding drug and drug-loading rate, and to explore the effect of ultrasonography (US) and contrast-enhanced ultrasonography (CEUS) on the imaging of HD@P-NPs in the presence of LIFU; logarithmically grown keloid fibroblasts KFS were taken and co-cultured with different experimental groups and divided into the following five groups: control, LIFU irradiation alone, D@P-NPs, HD@P-NPs, and HD@P-NPs+LIFU irradiation, and the acoustic kinetic efficacy of the nanoprobe HD@P-NPs was assessed using Calcein-AM/propidium iodide staining method, cell viability was detected by MTT assay, and the migration ability of KFS cells was studied and DCFH-DA indicator was applied to observe intracellular reactive oxygen species production. RESULTS ①The nanoprobes HD@P-NPs were prepared with uniform size and good stability, with an average particle size of 170.36±6.03nm, a potential of -36.91±3.56mv, and HMME encapsulation and drug loading rates of 67.41% and 5.18%, respectively, and DOX encapsulation and drug loading rates of 72.80% and 11.20%, respectively.②After LIFU irradiation, the phase transition of lipid nanoprobes HD@P-NPs produces microbubbles, and the maximum imaging effect can be achieved at 3W/cm2 and 2min.③MTT and Calcein-AM/propidium iodide staining showed that the nanoprobe HD@P-NPs had a significant cytostatic effect on KFS cells under combined LIFU conditions, which showed a clear red fluorescence, while LIFU irradiation alone did not show any cytostatic effect, the cytostatic effect was weaker in the D@P-NPs group and the HD@P-NPs group than in the HD@P-NPs+LIFU group, and there was no statistically significant difference in cytostatic inhibition between the two groups (P > 0.05). ④KFS cell migration assay showed that there was no statistically significant difference in cell migration inhibition between the D@P-NPs group and the HD@P-NPs group in the absence of LIFU (P＞0.05), while the HD@P-NPs+LIFU group showed the maximum cell migration inhibition, with a migration rate of only 10.32% (P < 0.05), and a significant increase of ROS production of the KFS cells in the presence of LIFU. Conclusion Lipid nanoprobes HD@P-NPs were successfully prepared, which can increase the effective drug concentration in the target area under the combined LIFU effect and realize the acoustic power cascade amplification therapy of KFS cells under ultrasound visualization. |
| Key words: Acoustic power therapy Lipid nanoprobe Keloid fibroblasts Hematoporphyrin monomethyl ether Adriamycin Ultrasonography |
