Small Cellular Particles from European Spruce Needle Homogenate

• Published in: International journal of molecular sciences Vol. 24; no. 5; p. 4349
• Main Authors: Jeran, Marko, Romolo, Anna, Spasovski, Vesna, Hočevar, Matej, Novak, Urban, Štukelj, Roman, Šuštar, Vid, Kisovec, Matic, Bedina Zavec, Apolonija, Kogej, Ksenija, Iglič, Aleš, Trebše, Polonca, Kralj-Iglič, Veronika
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 22.02.2023; MDPI
• Subjects: Alzheimer's disease; Antimicrobial agents; Deoxyribonucleic acid; DNA; Drug delivery systems; extracellular particles; extracellular vesicles; Flow Cytometry; Gas chromatography; Gas Chromatography-Mass Spectrometry; Gram-positive bacteria; interferometric light microscopy; Light microscopy; Mass spectrometry; Mass spectroscopy; Metabolites; Microscopy; Nanoparticles; Optical microscopy; Phenolic compounds; Picea; Picea abies; Pine needles; Scanning electron microscopy; small cellular particles; Spectrum analysis; spruce needles; Terpenes - analysis; Transmission electron microscopes; Ultracentrifugation; Vesicles; California; Slovenia; Massachusetts; Germany; spruce needles; cell-to-cell communication; extracellular particles; extracellular vesicles; exosomes; interferometric light microscopy; drug delivery; small cellular particles; Picea abies
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24054349

Small cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmission electron microscope (cryo TEM), assessed for their number density and hydrodynamic diameter by interferometric light microscopy (ILM) and flow cytometry (FCM), total phenolic content (TPC) by UV-vis spectroscopy, and terpene content by gas chromatography-mass spectrometry (GC-MS). The supernatant after ultracentrifugation at 50,000× contained bilayer-enclosed vesicles whereas in the isolate we observed small particles of other types and only a few vesicles. The number density of cell-sized particles (CSPs) (larger than 2 μm) and meso-sized particles (MSPs) (cca 400 nm-2 µm) was about four orders of magnitude lower than the number density of SCPs (sized below 500 nm). The average hydrodynamic diameter of SCPs measured in 10,029 SCPs was 161 ± 133 nm. TCP decreased considerably due to 5-day aging. Volatile terpenoid content was found in the pellet after 300× . The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use.