Baldini Nicola, Torreggiani Elena, Roncuzzi Laura, Perut Francesca, Zini Nicoletta, Avnet Sofia
Laboratory for Orthopaedic Pathophysiology and Regenerative Medicine, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy.
Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy.
Curr Pharm Biotechnol. 2018;19(11):877-885. doi: 10.2174/1389201019666181017115755.
Exosome-like nanovesicles are biological nanostructures mediating cell-tocell communication and capable to load selected cargos also in the interaction among different species.
We aimed to explore the content of exosome-like nanovesicles derived from Citrus limon L. and to analyze the effects of their uptake on human cells.
We isolated exosome-like nanovesicles from Citrus limon L. juice (EXO-CLs) by differential centrifugation. EXO-CLs were analyzed for short RNA content by advanced sequencing technologies, and for ascorbic acid (vitamin C) and citrate content by enzymatic assays. EXO-CLs anti-oxidant and pro-differentiative potential was evaluated in vitro on mesenchymal stromal cells (MSC), a common tool for regenerative strategies for several human tissues.
We showed that EXO-CLs carry detectable amounts of citrate and vitamin C and, although it was not possible to identify specific miRNAs, we detected short RNA sequences (20-30 bp) with unknown functions and with different distribution size in respect to whole Citrus limon L. juice. In vitro, EXO-CLs were uptaken by MSC and had a significant protective effect against oxidative stress. Furthermore, regarding the potential benefit for human bone health, we found that EXO-CLs modulate MSC differentiation versus the osteogenic lineage.
We demonstrated that incubation with EXO-CLs exert antioxidant activity in human cells. This is most likely due to the direct delivery and uptake of micronutrients by human cells that are well preserved inside the nanovesicle membrane, including the unstable vitamin C. Based on our results, we speculate that fruit-derived nanovesicles have the potential to mediate interspecies influence after food intake.
类外泌体纳米囊泡是介导细胞间通讯的生物纳米结构，在不同物种间的相互作用中也能够装载特定的货物。
我们旨在探索源自柠檬（Citrus limon L.）的类外泌体纳米囊泡的内容物，并分析其被人体细胞摄取后的影响。
我们通过差速离心从柠檬汁中分离出类外泌体纳米囊泡（EXO-CLs）。采用先进的测序技术分析EXO-CLs的短RNA含量，通过酶促测定法分析抗坏血酸（维生素C）和柠檬酸盐含量。在间充质基质细胞（MSC）上体外评估EXO-CLs的抗氧化和促分化潜力，MSC是多种人体组织再生策略的常用工具。
我们发现EXO-CLs携带可检测量的柠檬酸盐和维生素C，虽然无法鉴定出特定的微小RNA，但我们检测到了功能未知且与整个柠檬汁相比分布大小不同的短RNA序列（20-30 bp）。在体外，EXO-CLs被MSC摄取，并对氧化应激具有显著的保护作用。此外，关于对人体骨骼健康的潜在益处，我们发现EXO-CLs调节MSC向成骨谱系的分化。
我们证明与EXO-CLs共孵育可在人体细胞中发挥抗氧化活性。这很可能是由于人体细胞直接递送和摄取了纳米囊泡膜内保存完好的微量营养素，包括不稳定的维生素C。基于我们的结果，我们推测水果来源的纳米囊泡在摄入食物后有可能介导种间影响。
