Picciotto Sabrina, Barone Maria E, Fierli David, Aranyos Anita, Adamo Giorgia, BoŽič Darja, Romancino Daniele P, Stanly Christopher, Parkes Rachel, Morsbach Svenja, Raccosta Samuele, Paganini Carolina, Cusimano Antonella, Martorana Vincenzo, Noto Rosina, Carrotta Rita, Librizzi Fabio, Capasso Palmiero Umberto, Santonicola Pamela, Iglič Ales, Gai Meiyu, Corcuera Laura, Kisslinger Annamaria, Di Schiavi Elia, Landfester Katharina, Liguori Giovanna L, Kralj-Iglič Veronika, Arosio Paolo, Pocsfalvi Gabriella, Manno Mauro, Touzet Nicolas, Bongiovanni Antonella
Institute for Research and Biomedical Innovation (IRIB) - National Research Council (CNR), Palermo, Italy.
Centre for Environmental Research Innovation and Sustainability, Institute of Technology Sligo, Sligo, Ireland.
Biomater Sci. 2021 Apr 21;9(8):2917-2930. doi: 10.1039/d0bm01696a. Epub 2021 Feb 23.
Safe, efficient and specific nano-delivery systems are essential for current and emerging therapeutics, precision medicine and other biotechnology sectors. Novel bio-based nanotechnologies have recently arisen, which are based on the exploitation of extracellular vesicles (EVs). In this context, it has become essential to identify suitable organisms or cellular types to act as reliable sources of EVs and to develop their pilot- to large-scale production. The discovery of new biosources and the optimisation of related bioprocesses for the isolation and functionalisation of nano-delivery vehicles are fundamental to further develop therapeutic and biotechnological applications. Microalgae constitute sustainable sources of bioactive compounds with a range of sectorial applications including for example the formulation of health supplements, cosmetic products or food ingredients. In this study, we demonstrate that microalgae are promising producers of EVs. By analysing the nanosized extracellular nano-objects produced by eighteen microalgal species, we identified seven promising EV-producing strains belonging to distinct lineages, suggesting that the production of EVs in microalgae is an evolutionary conserved trait. Here we report the selection process and focus on one of this seven species, the glaucophyte Cyanophora paradoxa, which returned a protein yield in the small EV fraction of 1 μg of EV proteins per mg of dry weight of microalgal biomass (corresponding to 10 particles per mg of dried biomass) and EVs with a diameter of 130 nm (mode), as determined by the micro bicinchoninic acid assay, nanoparticle tracking and dynamic light scattering analyses. Moreover, the extracellular nanostructures isolated from the conditioned media of microalgae species returned positive immunoblot signals for some commonly used EV-biomarkers such as Alix, Enolase, HSP70, and β-actin. Overall, this work establishes a platform for the efficient production of EVs from a sustainable bioresource and highlights the potential of microalgal EVs as novel biogenic nanovehicles.
安全、高效且特异的纳米递送系统对于当前及新兴的治疗方法、精准医学和其他生物技术领域至关重要。基于细胞外囊泡（EVs）开发的新型生物基纳米技术近来不断涌现。在此背景下，确定合适的生物体或细胞类型作为可靠的EVs来源并开展其从小规模到大规模的生产变得至关重要。发现新的生物来源以及优化用于纳米递送载体分离和功能化的相关生物过程对于进一步开发治疗和生物技术应用至关重要。微藻是生物活性化合物的可持续来源，具有一系列行业应用，例如用于配制健康补充剂、化妆品或食品成分。在本研究中，我们证明微藻是有前景的EVs生产者。通过分析18种微藻产生的纳米级细胞外纳米物体，我们鉴定出7种有前景的产EVs菌株，它们属于不同的谱系，这表明微藻中产EVs是一种进化保守的特性。在此我们报告选择过程，并重点关注这7个物种之一，即蓝藻门的蓝氏原绿藻，通过微量双辛可宁酸测定、纳米颗粒跟踪和动态光散射分析确定，其小EV部分的蛋白质产量为每毫克微藻生物质干重1μg EV蛋白质（相当于每毫克干燥生物质10个颗粒），且EVs直径为130nm（众数）。此外，从微藻物种的条件培养基中分离出的细胞外纳米结构对一些常用的EV生物标志物（如Alix、烯醇化酶、热休克蛋白70和β-肌动蛋白）的免疫印迹信号呈阳性。总体而言，这项工作建立了一个从可持续生物资源高效生产EVs的平台，并突出了微藻EVs作为新型生物纳米载体的潜力。
