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Understanding the underlying transport mechanism of biological delivery is important for developing delivery technologies for pharmaceuticals, imaging agents, and nanomaterials. Recently reported by our group, SDots are a novel class of nanoparticle delivery systems with distinct biointerface features and excellent fusogenic capabilities (i.e., strong ability to interact with the hydrophobic portions of biomembranes). In this study, we investigate the cellular transport mechanism of SDots conjugated with Tat peptide (SDots-Tat) by live-cell spinning-disk confocal microscopy combined with molecular biology methods. Mechanistic studies were conducted on the following stages of cellular transport of SDots-Tat in HeLa cells: cellular entry, endosomal escape, nucleus entry, and intranuclear transport. A key finding is that, after escaping endosomes, SDots-Tat enter the cell nucleus via an importin β-independent pathway, bypassing the usual nucleus entry mechanism used by Tat. This finding implies a new approach to overcome the nucleus membrane barrier for designing biological delivery technologies.
Introduction: Bacterial pneumonia poses a significant global public health challenge, where unaddressed pathogens and inflammation can exacerbate acute lung injury and prompt cytokine storms, increasing mortality rates. Alveolar macrophages are pivotal in preserving lung equilibrium. Excessive inflammation can trigger necrosis in these cells, disrupting the delicate interplay between inflammation and tissue repair. Methods: We obtained extracellular vesicle from aloe and tested the biosafety by cell viability and hemolysis assays. Confocal microscopy and flow cytometry were used to detect the uptake and internalization of extracellular vesicle by macrophages and the ability of extracellular vesicle to affect the phenotypic reprogramming of macrophages in vitro. Finally, we conducted a clinical feasibility study employing clinical bronchoalveolar lavage fluid as a representative model to assess the effective repolarization of macrophages influenced by extracellular vesicle. Results: In our study, we discovered the potential of extracellular vesicle nanovesicles derived from aloe in reprograming macrophage phenotypes. Pro-inflammatory macrophages undergo a transition toward an anti-inflammatory immune phenotype through phagocytosing and internalizing these aloe vera-derived extracellular vesicle nanovesicles. This transition results in the release of anti-inflammatory IL-10, effectively curbing inflammation and fostering lung tissue repair. Discussion: These findings firmly establish the immunomodulatory impact of aloe-derived extracellular vesicle nanovesicles on macrophages, proposing their potential as a therapeutic strategy to modulate macrophage immunity in bacterial pneumonia.
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