No abstract

Endodontic persistent infections are often mediated by bacterial biofilms. This mode of bacterial growth is characterized by the presence of a matrix mainly composed of extracellular polymeric substances (EPSs) that protect the encased microorganisms. To establish better control and disinfection protocols, elucidation of the main components of biofilm matrices present in endodontic infections is required. The aim of the present study was to characterize the principal components of E. faecalis, A. naeslundii, and dual-species biofilm matrices by means of Raman spectroscopy and confocal scanning laser microscopy (CSLM) techniques. The total biomass of biofilms was quantified via crystal violet assays, and the monospecies biofilms showed higher biomass than the dual-species biofilms. Raman spectroscopy and confocal laser scanning microscopy were used to identify the biochemical composition and structure of the biofilm matrices. Spectra originating from the biofilms of two endodontic pathogens show the presence of carbohydrates, proteins, fatty acids, and nucleic acids in all samples; however, variation in the levels of expression of these biomolecules allows spectroscopic differentiation of the biofilms using principal component analysis. This study is the first attempt to identify the composition of monospecies and dual-species biofilms of endodontic origin. Our data provides an important approach to the understanding of molecular dynamics of endodontic infections.

Apical periodontitis is frequently associated with the presence of bacteria biofilm, which has an indisputable impact on the prognosis of endodontic therapy due to the high resistance to adverse environmental conditions, chemicals, and antibiotic therapy that characterize bacteria within biofilm. The biofilm matrix acts as a protective shield over the encased microorganisms. The aim of this investigation was to identify the main biochemical components of biofilm matrix from endodontic mono- and dual-species biofilms. Enterococcus faecalis and Actinomyces naeslundii were cultured as mono- and dual-species biofilms for 14 days. Crude extracellular polymeric substances (EPSs) from biofilm matrices were extracted using chemical and physical methods. High-performance liquid chromatography, gas chromatography, and mass spectrometry were used to determine the carbohydrate, protein, and fatty acid components. Chemical analysis of the biofilm matrices revealed that they were mainly composed of stachyose, maltose, and mannose carbohydrates. The protein profile in all biofilm samples showed abundant oxidoreductases and chaperone proteins and some virulence- associated proteins mainly located in the membrane surface. High percentages of saturated and monounsaturated fatty acids were identified in all biofilm matrices, with a major prevalence of palmitic, stearic, and oleic acids. Based on the results, it was possible to obtain for the first time a general overview of the biochemical profile of endodontic biofilm matrices.

No abstract

Diatomite samples from Costa Rica were purified using acidic treatments with hydrochloric acid, thermally treated (400-1000°C) and then silylated with trimethylchlorosilane in toluene under inert atmosphere. The purification process allows to decrease the concentration of metals presented in the crude diatomite, as is confirmed by X-ray Fluorescence (XRF) Analysis. The silylated materials were analyzed by using Hyperpolarized 129 Xe Nuclear Magnetic Resonance Spectroscopy (HP 129 Xe NMR), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Thermogravimetric Analysis (TGA), rehydration tests, and contact angle measurements. XRD measurements indicate that diatomite is mainly amorphous, but presents several crystalline phases (kaolinite, cristobalite, and quartz). Pretreatments at high temperatures cause changes in those crystalline phases, resulting in more amorphous materials. However, there is no difference in the overall structure of purified and thermally treated diatomite samples with respect to the silylation products. In addition, SEM measurements show no effect over the pore structure of the materials. On the other hand, TGA measurements and rehydration tests show lower losses of water for silylated materials prepared using higher pretreatment temperatures. Moreover, HP 129 Xe NMR, FTIR, and contact angle measurements evidence a modification due to covalent attachment of Si(CH 3 ) 3 -groups to the surface, which increases for higher pretreatment temperatures. The results provide valuable information about external factors that influence the surface modification of diatomite. This can be useful to control modifications that can be achieved in a similar way.