Monitoring of changes in the concentrations of the low molecular weight constituents enhanced by abundant proteins depletion.

No abstract

Raman spectroscopy can provide a molecular-level fingerprint of the biochemical composition and structure of cells with excellent spatial resolution and could be useful to monitor changes in composition for dysplasia and early, non-invasive cancer diagnosis (carcinoma in situ), both ex-vivo and in vivo. In this study, we demonstrate this potential by collecting Raman spectra of the nucleoli, nuclei and cytoplasm from oral epithelial cancer (SCC-4) and dysplastic (pre-cancerous, DOK) cell lines and from normal oral epithelial primary cell cultures, in vitro, which were then analysed by principal component analysis (PCA) as a multivariate statistical method to discriminate the spectra. Results show significant discrimination between cancer and normal cell lines. Furthermore, the dysplastic and cancer cell lines could be discriminated based on the spectral profiles of the cytoplasmic regions. The principal component loading plot, which elucidates the biochemical features responsible for the discrimination, showed significant contributions of nucleic acid and proteins for nucleolar and nuclear sites and variation in features of lipids for the cytoplasmic area. This technique may provide a rapid screening method and have potential use in the diagnosis of dysplasia and early, non-invasive oral cancer, the treatment of which involves much less extensive and complex surgery and a reduction in associated co-morbidity for the patient.

The present study aimed to evaluate the erosive potential of four most commonly prescribed syrup medicaments for respiratory diseases. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy combined with multivariate statistical analysis and micro-energy-dispersive X-ray fluorescence spectrometry (μ-EDXRF) mapping was performed. Fifty-five root dentin fragments obtained from the buccal surface of 30 bovine teeth were prepared and divided into five experimental groups (n = 10): control-artificial saliva (S), acebrofilin hydrochloride (AC), ambroxol hydrochloride (AM), bromhexine hydrochloride (BR), and salbutamol sulfate (SS). The S group was stored only in artificial saliva and the other groups were treated with the medicaments (immersed for 1 min in 3 mL of the medication, three times daily, with 1-h intervals between the immersion cycles, during 5 days, 15 immersion cycles). There were a significant decrease in the Ca and P weight percentages (wt%) for dentin after medication treatments, except for AC (p > 0.05). Mineral content of dentin showed a clear gradation with increasing Ca and P wt% reduction in the order S < AC < AM < BR < SS. SS resulted in a significant increase in Ca/P ratio when compared to the control (p < 0.001). ATR-FTIR combined with multivariate, statistical analysis can quickly and reliably indicate extent of dentin erosion. Considering syrups with high-erosive potential should always follow with proper oral hygiene practices or search for an alternative medications void of such detrimental effects. Regular and prolonged use of these medicaments might bear the risk of causing erosion.

Raman spectroscopy can provide a molecular-level fingerprint of the biochemical

Objectives Investigate the biochemistry of in vivo healthy oral tissues through Raman spectroscopy. We aimed to characterize the biochemical features of healthy condition in oral subsites (buccal mucosa, lip, tongue, and gingiva) of healthy subjects. More specifically, we investigated Raman spectral characteristics and biochemical content of in vivo healthy tissues on Brazilian population. This characterization can be used to better define normal tissue and improve the detection of oral premalignant conditions in future studies. Materials and methods For spectroscopic analysis a Raman spectrometer (Kaiser Optical Systems imaging spectrograph Holospec, f / 1.8i-NIR) coupled with a laser 785 nm, 60 mW was used. Raman measurements were obtained by means of an optical fiber (EMVision fiber optic probe) coupled between the laser and the spectrometer. Three spectra per site were acquired from the lip, buccal mucosa, tongue, and gingiva of ten healthy volunteers. This resulted in 30 spectra per oral sub-site and in total 120 spectra.Results We report detailed biochemical information on these subsites and their relative composition based on deconvolution studies of their spectra. Finally, we also report classification efficiency of 61, 83, 41, and 93% for buccal, gingiva, lip, and tongue respectively after applying multivariate statistical tools. Conclusions We quantitated the contribution of various biochemicals in terms of percentage, and this will enable comparison not only across anatomical sites but also across studies. Raman spectroscopy can rapidly probe tissue biochemistry of healthy oral regions. Moreover, the study suggests the possibility of using Raman spectroscopy combined with signal processing and multivariate analysis methods to differentiate the oral sites in healthy conditions and compare with pathological conditions in future studies. Clinical relevance The spectral characterization of the healthy condition of oral tissues by a noninvasive, label-free, and real-time analytical techniques is important to create a spectral reference for future diagnosis of pathological conditions.

The present study aims to evaluate the effect of brushing with fluoride dentifrice on teeth severely affected by erosion due to respiratory medicaments. Enamel (n = 50) and dentin (n = 50) bovine specimens were prepared and treated with artificial saliva (S‐control), acebrofilin hydrochloride (AC), ambroxol hydrochloride (AM), bromhexine hydrochloride (BR), and salbutamol sulfate (SS) and subjected to cycles of demineralization (immersing in 3 mL, 1 min, three times a day at intervals of 1 hr, for 5 days) followed by remineralization (saliva, 37°C, 1 hr). Simulated brushing with fluoridated toothpaste was performed using 810 strokes in a reciprocal‐action brushing simulator. Scanning electron microscopy, micro energy dispersive X‐ray fluorescence (μ‐EDXRF) spectroscopy and attenuated total reflection Fourier transform infrared (ATR FTIR) spectroscopy were then performed. μ‐EDXRF images showed extensive erosion after treatment with all medicaments. SEM images showed enamel erosion in order SS > BR > AC = AM > S after brushing and fluoridation. FTIR results were in agreement. In case of dentin, μ‐EDXRF measurements showed significant difference in mineral content (percent weight of calcium and phosphate) in SS + brushing + fluoridation treated enamel compared to control, while μ‐EDXRF images showed erosive effects in the order SS > AM>BR > AC = S post brushing + fluoridation. SEM images showed erosion in the order SS > AM = BR > AC > S post brushing + fluoridation. Again, FTIR multivariate results were in agreement. Overall, our study shows that proper oral care is critical when taking certain medication. The study also demonstrates the possible use of FTIR for rapid clinical monitoring of tooth erosion in clinics.

Raman spectroscopy can provide a molecular-level signature of the biochemical composition and structure of cells with submicrometer spatial resolution and could be useful to monitor changes in composition for early stage and non-invasive cancer diagnosis, both ex-vivo and in vivo. In particular, the fingerprint spectral region (400-1800cm) has been shown to be very promising for optical biopsy purposes. However, limitations for discrimination of dysplastic and inflammatory processes based on the fingerprint region have been demonstrated. In addition, the Raman spectral signal of dysplastic cells is one important source of misdiagnosis of normal versus pathological tissues. The high wavenumber region (2800-3600cm) provides more specific information based on NH, OH and CH vibrations and can be used to identify the subtle changes which could be important for discrimination of samples. In this study, we demonstrate the potential of the high-wavenumber spectral region in this context by collecting Raman spectra of nucleolus, nucleus and cytoplasm from oral epithelial cancer (SCC-4) and dysplastic (DOK) cell lines and from normal oral epithelial primary cells, in vitro, in water immersion, which were then analyzed by principal components analysis as a method to discriminate the spectra. Analysis was performed before and after digital subtraction of the bulk water signal. In the normal cell line, the three subcellular regions are well differentiated before water subtraction, although the discrimination of the two nuclear regions is less well defined after water subtraction. Comparing the respective subcellular regions of the three cell lines, before water subtraction, the cell lines can be discriminated using sequential PCA and Feature Discriminant Analysis with up to ~100% sensitivity and 97% specificity for the cytoplasm, which is improved to 100% sensitivity and 99% specificity for the nucleus. The results are discussed in terms of discrimination comparing the CH vibrational modes of nucleic acids, proteins and lipids. The potential role of the OH vibrations, considering free water and confined water, in the discrimination of cell cultures and pathological processes are also discussed.