While microRNAs are considered as excellent biomarkers of various diseases, there are still several remaining challenges regarding their isolation. In this study, we aimed to design a novel RNA isolation method that would help to overcome those challenges. Therefore, we present a novel phenol/chloroform-free, low-cost method for miRNA extraction. Within this method, RNA is extracted from cell lysate with an isopropanol/water/NaCl system, followed by solid-phase extraction using TiO2 microspheres to effectively separate short RNAs from long RNA molecules. We also demonstrated the pH-dependent selectivity of TiO2 microspheres towards different sizes of RNA. We were able to regulate the size range of extracted RNAs with simple adjustments in binding conditions used during the solid-phase extraction.

• Klíčová slova
• TiO2, miRNA isolation, phenol/chloroform-free, short RNA extraction,
• MeSH
• chloroform chemie   MeSH
• fenol * chemie   MeSH
• fenoly MeSH
• mikro RNA * genetika   MeSH
• titan MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chloroform MeSH
• fenol * MeSH
• fenoly MeSH
• mikro RNA * MeSH
• titan MeSH
• titanium dioxide MeSH Prohlížeč

The immunoreactivity or/and stress response can be induced by nanomaterials' different properties, such as size, shape, etc. These effects are, however, not yet fully understood. This study aimed to clarify the effects of SiO2 nanofibers (SiO2NFs) on the cellular responses of THP-1-derived macrophage-like cells. The effects of SiO2NFs with different lengths on reactive oxygen species (ROS) and pro-inflammatory cytokines TNF-α and IL-1β in THP-1 cells were evaluated. From the two tested lengths, it was only the L-SiO2NFs with a length ≈ 44 ± 22 µm that could induce ROS. Compared to this, only S-SiO2NFs with a length ≈ 14 ± 17 µm could enhance TNF-α and IL-1β expression. Our results suggested that L-SiO2NFs disassembled by THP-1 cells produced ROS and that the inflammatory reaction was induced by the uptake of S-SiO2NFs by THP-1 cells. The F-actin staining results indicated that SiO2NFs induced cell motility and phagocytosis. There was no difference in cytotoxicity between L- and S-SiO2NFs. However, our results suggested that the lengths of SiO2NFs induced different cellular responses.

• Klíčová slova
• SiO2 nanofibers, THP-1-derived macrophage-like cells, cytotoxicity, immunoreactivity,
• MeSH
• cytokiny metabolismus   MeSH
• lidé MeSH
• makrofágy MeSH
• oxid křemičitý * farmakologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• THP-1 buňky MeSH
• TNF-alfa * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytokiny MeSH
• oxid křemičitý * MeSH
• reaktivní formy kyslíku MeSH
• TNF-alfa * MeSH