REMEDI at CÚRAM Centre for Medical Device Research, University of Galway, Galway, Ireland.
Orbsen Therapeutics, IDA Business Park, Dangan, Galway, Ireland.
Stem Cell Res Ther. 2023 Jun 6;14(1):151. doi: 10.1186/s13287-023-03385-6.
Mesenchymal stem cell (MSC) derived extracellular vesicles (EVs) have been proposed as an alternative to cell therapy, creating new possible delivery modalities such as nebulisation. We wished to investigate the therapeutic potential of directly nebulised MSC-EVs in the mitigation of Escherichia coli-induced pneumonia.
EV size, surface markers and miRNA content were assessed pre- and post-nebulisation. BEAS2B and A459 lung cells were exposed to lipopolysaccharide (LPS) and treated with nebulised bone marrow (BM) or umbilical cord (UC) MSC-EVs. Viability assays (MTT) and inflammatory cytokine assays were performed. THP-1 monocytes were stimulated with LPS and nebulised BM- or UC-EVs and phagocytosis activity was measured. For in vivo experiments, mice received LPS intratracheally (IT) followed by BM- or UC-EVs intravenously (IV) and injury markers assessed at 24 h. Rats were instilled with E. coli bacteria IT and BM- or UC-EVs delivered IV or by direct nebulisation. At 48 h, lung damage was assessed by physiological parameters, histology and inflammatory marker presence.
MSC-EVs retained their immunomodulatory and wound healing capacity after nebulisation in vitro. EV integrity and content were also preserved. Therapy with IV or nebulised MSC-EVs reduced the severity of LPS-induced lung injury and E. coli-induced pneumonia by reducing bacterial load and oedema, increasing blood oxygenation and improving lung histological scores. MSC-EV treated animals also showed lower levels of inflammatory cytokines and inflammatory-related markers.
MSC-EVs given IV attenuated LPS-induced lung injury, and nebulisation of MSC-EVs did not affect their capacity to attenuate lung injury caused by E. coli pneumonia, as evidenced by reduction in bacterial load and improved lung physiology.
间充质干细胞(MSC)衍生的细胞外囊泡(EVs)已被提议作为细胞治疗的替代方法,创造了新的可能的递药方式,如雾化。我们希望研究直接雾化 MSC-EVs 在减轻大肠杆菌诱导的肺炎中的治疗潜力。
在雾化前后评估 EV 的大小、表面标志物和 miRNA 含量。BEAS2B 和 A459 肺细胞暴露于脂多糖(LPS)并用雾化骨髓(BM)或脐带(UC)MSC-EVs 处理。进行细胞活力测定(MTT)和炎症细胞因子测定。用 LPS 刺激 THP-1 单核细胞并用雾化 BM 或 UC-EVs 处理,并测量吞噬活性。在体内实验中,小鼠接受 LPS 气管内(IT)注射,然后静脉内(IV)给予 BM 或 UC-EVs,并在 24 小时评估损伤标志物。大鼠接受 IT 大肠杆菌细菌感染并用 IV 或直接雾化给予 BM 或 UC-EVs。在 48 小时时,通过生理参数、组织学和炎症标志物存在来评估肺损伤。
MSC-EVs 在体外雾化后仍保留其免疫调节和伤口愈合能力。EV 的完整性和内容也得到了保留。IV 或雾化 MSC-EVs 治疗可通过降低细菌负荷和水肿、增加血氧饱和度和改善肺组织学评分来减轻 LPS 诱导的肺损伤和大肠杆菌诱导的肺炎的严重程度。MSC-EV 处理的动物还表现出较低水平的炎症细胞因子和炎症相关标志物。
IV 给予 MSC-EVs 可减轻 LPS 诱导的肺损伤,而雾化 MSC-EVs 不会影响其减轻大肠杆菌性肺炎引起的肺损伤的能力,这表现为细菌负荷降低和肺生理改善。