Biofilms are a communal way of living for microorganisms in which microorganism cells are surrounded by extracellular polymeric substances (EPS). Most microorganisms can live in biofilm form. Since microorganisms are everywhere, understanding biofilm structure and composition is crucial for making the world a better place to live, not only for humans but also for other living creatures. Raman spectroscopy is a nondestructive technique and provides fingerprint information about an analyte of interest. Surface-enhanced Raman spectroscopy is a form of this technique and provides enhanced scattering of the analyte that is in close vicinity of a nanostructured noble metal surface such as silver or gold. In this review, the applications of both techniques and their combination with other biofilm analysis techniques for characterization of composition and structure of biofilms are discussed.
Citrate reduced colloidal silver nanoparticles (c-AgNPs) as synthesized and modified with oligonucleotides (Oligo-AgNPs) are comparatively evaluated for their wound healing properties on animal models. The healing progress was monitored daily during nine days by measuring the wound diameter. The tissue samples from the healed regions were analyzed for epithelial damage, congestion, inflammatory cell infiltration, fibroblast proliferation, and new collagen synthesis. The c-AgNPs and Oligo-AgNPs had statically significant impact on the healing process compared to control. The histological analysis revealed that the c-AgNPs and Oligo-AgNPs improved the congestion, inflammatory cell infiltration, fibroblast proliferation and new collagen synthesis as compared to control. Although the fibroblast proliferation seems to be the same for both c-AgNPs and Oligo-AgNPs, the collagen synthesis is markedly improved with the Oligo-AgNPs. The atomic spectroscopy analysis of the samples from different tissues showed that the AgNPs applied topically to the skin does not pass through the other organs. Our data suggest that topical application of OligosAgNPs improve wound healing by promoting increased collagen synthesis and tissue re-modeling without any side effects.
Biofilm formation is a defense mechanism for microorganisms to survive under both natural and stress conditions. Clinically relevant microorganisms threaten patient health through biofilm formation on medical devices and implants. It is very important to identify biofilm formation in order to suppress their pathogenic activities in early stages. With the aim for better understanding biofilm formation and possibility of detection, in this study, biofilm formation of clinically important microorganisms, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Candida albicans are monitored with surface-enhanced Raman scattering (SERS). The SERS spectra were collected by mapping a dried droplet area where a volume of colloidal silver nanoparticle (AgNP) suspension is placed on microorganism culture plate. The spectral changes on the SERS spectra with increasing incubation time of the model microorganisms from 4 to 120 h are monitored. The unique spectra originating from the biofilms of three pathogenic microorganisms and the spectral changes as a result of time-dependent concentration fluctuations of biomolecular species in their biofilms including carbohydrates, lipids, proteins, and genetic materials allow not only identification but also discrimination of biofilms using principal component analysis.
In recent years, an increasing number of research papers revealed that the compositional and volumetric alterations in the extracellular matrix are the consequences of aging and may be related to Alzheimer's disease (AD). In this study, we aimed to demonstrate the alterations in hippocampal extracellular fluid proteins in vivo using the 5XFAD mouse model. Samples were obtained from hippocampi of 5XFAD mice (n = 6) and their non-transgenic littermates by intracerebral push-pull perfusion technique at 3 months of age, representing the pre-pathological stage of the AD. Proteins in the hippocampal perfusates were analyzed by Ultra Performance Liquid Chromatography-Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry (UPLC-ESI-qTOF-MS/MS). 178 proteins were identified and 19 proteins of them were found to be statistically significantly altered (p≤0.05, fold change ≥40%, unique peptide count ≥3) in the hippocampal CA1 extracellular fluid of the 5XFAD mouse model. Ingenuity pathway analysis of the protein expression results identified IL6 as an upstream regulator. The upregulation of IL6 was validated by immunohistochemical staining of the hippocampus and cortex of the 5XFAD mice prior to Aβ plaque formation. Furthermore, the iron level in the hippocampus was measured by inductively coupled plasma-mass spectrometry as IL6 is mentioned in several studies to take part in iron homeostasis and inflammation and found to be increased in 5XFAD mice hippocampus. Alterations in extracellular matrix proteins in addition to increasing amount of hippocampal IL6 and iron in the early stages of AD may reveal inflammation-mediated iron dyshomeostasis in the early stages of neurodegeneration.
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