Extraction of Extracellular Matrix in Static and Dynamic Candida Biofilms Using Cation Exchange Resin and Untargeted Analysis of Matrix Metabolites by Ultra-High-Performance Liquid Chromatography-Tandem Quadrupole Time-of-Flight Mass Spectrometry (UPLC-Q-TOF-MS)

This study aims to investigate the efficiencies of seven methods in C. albicans biofilm EM extraction, namely vortexing plus ultrasonication (VU), formaldehyde plus vortexing and ultrasonication (Formaldehyde-VU), EDTA plus vortexing and ultrasonication (EDTA-VU), NaOH plus vortexing and ultrasonication (NaOH-VU), ethanol plus vortexing and ultrasonication (Ethanol-VU), ion exchange resin plus vortexing and ultrasonication (IER-VU)—including cation/anion-exchange resin plus vortexing and ultrasonication (CER/AER-VU)—under the static and dynamic states. Subsequently, the extracted metabolites are analyzed with the UPLC-Q-TOF-MS tool via untargeted filtration. Materials and Methods Strains and Cultivation Candida albicans SC5314 was kindly provided by Prof. Yuanying Jiang from the College of Pharmacy, the Second Military Medical University (Shanghai, China). All the stock cultures of these strains were conventionally preserved in Sabouraud agar and propagated in liquid Sabouraud medium (Hope Biotech, Co., Qingdao, China) at 37°C for 12–16 h when the cells reached the exponential phase. The revived Candida cells were collected at 5200 rpm (Leiboer Medical Devices, Beijing, China) and washed twice with sterile phosphate-buffered saline (PBS, Leagene, Beijing, China). The fungal cells were then resuspended in RPMI-1640 medium (Invitrogen, Carlsbad, CA, United States) and adjusted to a defined cell density using a hemocytometer prior to the following ...
Source: Frontiers in Microbiology - Category: Microbiology Source Type: research