Brun Edgar Hernan Cuevas, Hong Zuo-Yi, Hsu Yuan-Ming, Wang Ciou-Ting, Chung Dai-Jung, Ng Shang-Kok, Lee Yau-Hsuan, Wei Tzu-Tang
HCmed Innovations Co., Ltd., Taipei, Taiwan.
Department and Graduate Institute of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan.
J Aerosol Med Pulm Drug Deliv. 2023 Apr;36(2):55-64. doi: 10.1089/jamp.2022.0020. Epub 2023 Feb 23.
Idiopathic pulmonary fibrosis (IPF) is a serious lung disease characterized by lung scarring, which results in breathing difficulty. Currently, patients with IPF exhibit a poor survival rate and have access to very limited therapeutic options. Interferon beta (IFN-β) has been approved by the U.S. Food and Drug Administration (FDA) for the treatment of relapsing forms of multiple sclerosis, and it has also been shown to exhibit therapeutic potential in IPF. However, clinical use of IFN-β did not lead to improved overall survival in IPF patients in existing studies. One possibility is the limited efficiency of IFN-β delivery through intravenous or subcutaneous injection. The aerosol particle size distribution was determined with a laser diffraction particle size analyzer to characterize the droplet size and fine particle fraction generated by three types of nebulizers: jet, ultrasonic, and mesh. A breathing simulator was used to assess the delivery efficiency of IFN-β, and the temperature in the medication reservoirs was monitored with a thermocouple during nebulization. To further evaluate the antifibrotic activity of IFN-β pre- and postnebulization, bleomycin (BLM)- or transforming growth factor-beta (TGF-β)-treated human lung fibroblast (HLF) cells were used. Cell viability was measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Transwell migration assay and Q-PCR analysis were used to evaluate cell migration and the myofibroblast differentiation ability, respectively. IFN-β protein samples were prepared using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample loading buffer, and the expression of IFN-β was assessed by western blotting. Among the current drug delivery systems, aerosolized medication has shown increased efficacy of drug delivery for treating respiratory diseases when compared with parenteral drugs. It was found that neither the structural integrity nor the biological function of nebulized IFN-β was compromised by the nebulization process of the mesh nebulizer. In addition, in BLM dose-response or TGF-β-induced lung fibroblast proliferation assays, these effects could be reversed by both parenteral and inhaled IFN-β nebulized with the mesh nebulizer. Nebulized IFN-β with the mesh nebulizer also significantly inhibited the migration and myofibroblast differentiation ability of TGF-β-treated HLF cells. The investigations revealed the potential efficacy of IFN-β in the treatment of IPF with the mesh nebulizer, demonstrating the higher efficiency of IFN-β delivered through the mesh nebulizer.
特发性肺纤维化（IPF）是一种以肺瘢痕形成为特征的严重肺部疾病，可导致呼吸困难。目前，IPF患者的生存率较低，且治疗选择非常有限。干扰素β（IFN-β）已被美国食品药品监督管理局（FDA）批准用于治疗复发型多发性硬化症，并且在IPF中也显示出治疗潜力。然而，在现有研究中，IPF患者使用IFN-β进行临床治疗并未提高总体生存率。一种可能性是通过静脉或皮下注射递送IFN-β的效率有限。使用激光衍射粒度分析仪测定气溶胶粒度分布，以表征三种雾化器（喷射式、超声式和网式）产生的液滴大小和细颗粒分数。使用呼吸模拟器评估IFN-β的递送效率，并在雾化过程中用热电偶监测药物储器中的温度。为了进一步评估雾化前后IFN-β的抗纤维化活性，使用了博来霉素（BLM）或转化生长因子-β（TGF-β）处理的人肺成纤维细胞（HLF）。通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑（MTT）法测定细胞活力。分别使用Transwell迁移试验和Q-PCR分析评估细胞迁移和成肌纤维细胞分化能力。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）上样缓冲液制备IFN-β蛋白样品，并通过蛋白质印迹法评估IFN-β的表达。在目前的药物递送系统中，与肠胃外给药相比，雾化药物在治疗呼吸系统疾病方面显示出更高的药物递送效率。结果发现，网式雾化器的雾化过程不会损害雾化IFN-β的结构完整性或生物学功能。此外，在BLM剂量反应或TGF-β诱导的肺成纤维细胞增殖试验中，肠胃外给药和用网式雾化器雾化吸入的IFN-β均可逆转这些作用。用网式雾化器雾化的IFN-β还显著抑制了TGF-β处理的HLF细胞的迁移和成肌纤维细胞分化能力。这些研究揭示了网式雾化器递送IFN-β治疗IPF的潜在疗效，证明了通过网式雾化器递送IFN-β的效率更高。
