Dunham, KaitlynCanham, SpencerPalmer, EmilieBai, ShuhuaYang, Tianzhi2023-11-132023-11-132023-04-20https://dspace.husson.edu/handle/20.500.14298/56Blueberries provide evident benefits of reducing oxidative stress and inflammation, improving cognitive function, and protecting against neurodegenerative diseases as a “super” fruit. However, high levels of bioactive molecules in blueberries, such as anthocyanin, have poor stability and absorption, leading to low bioavailability in the brain. Exosomes attract strong interest as an important vehicle of intercellular communication and as a delivery carrier of bioactive molecules. Herein, the study focuses on characterizing blueberry-derived exosomal nano-vesicles that contain bioactive molecules, and determining if they have better stability and are more readily taken up by cells. Blueberry juice was directly extracted with an electric blender and passed through filter papers. The collected juice was sequentially centrifuged at 1,000 × g for 10 min, 3,000 × g for 20 min, and 10,000 × g for 40 min at 4 °C to remove large particles and debris. The final supernatant was centrifuged at 100,000 × g for 30 min to obtain exosomes. The exosome morphology was observed with scanning electron microscopy (SEM) and the particle size was determined with a Nano Sizing Analyzer. The exosomes had a size of 82.7±6.4 nm and appeared as individually sphere-shaped morphology as shown in SEM images. Exosomes were analyzed and quantified for total proteins and RNAs. Well-known exosome-unique markers, including Cis-Golgi matrix protein GM130, adaptor protein and sort cargo ALIX, tumor susceptibility gene TSG10, intercellular adhesion molecule 1 (ICAM-1), apoptosis ANXA5, integral membrane protein FLOT, epithelial cell adhesion molecule (EpCam), and transmembrane tetraspanin family CD63 and CD81, were detected by an Exo-Check array. MicroRNA sequencing analysis revealed that the intersection between differentially expressed genes and miRNAs contained in exosomes could unveil a set of candidate target genes. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) study demonstrated that the exosomes showed cytotoxicity in neural cancerous cells. The characterized exosomes with biomolecules may deliver therapeutic molecules in the brain and target neural cells, leading to improved efficacy in the treatment of neurodegenerative diseases.en-USAbstract