In vivo Raman spectroscopic characteristics of different sites of the oral mucosa in healthy volunteers.

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.

Effect of non-thermal atmospheric plasma on the dentin-surface topography and composition and on the bond strength of a universal adhesive.

This study investigated the effect of application of non-thermal atmospheric plasma (NTAP) on the topography and composition of the dentin surface, as well as the microtensile bond strength (µTBS) of a universal adhesive to NTAP-treated dentin. Exposed flat dentin surfaces from human third molars were either treated with NTAP for 10 and 30 s or untreated (control). The dentin-surface topography and chemical composition were characterized by atomic force microscopy (n = 3) and Raman confocal spectroscopy (n = 5), respectively. The µTBS (n = 8) of Scotchbond Universal to dentin was determined after storage for 24 h and 1 yr, either by direct water exposure or under simulated pulpal pressure. In-situ zymography was used to evaluate the influence of NTAP on the dentin-enzymatic activity. Non-thermal atmospheric plasma produced no remarkable topographical or chemical alterations at the dentin surface; only the amount of phosphate decreased following 10 s of treatment with NTAP. After 1 yr of direct water exposure, the µTBS of NTAP-treated specimens did not differ statistically significantly from that of untreated controls, whereas simulated pulpal pressure-aging resulted in a significantly higher µTBS for NTAP-treated dentin. The dentin-enzymatic activity appeared to be treatment-dependent, but the untreated controls showed more intense fluorescence within the hybrid layer. Scotchbond Universal maintained its µTBS strength after 1 yr of direct water exposure and simulated pulpal pressure, although remarkable statistical differences between treatments were observed depending on the aging condition.

Effects of heating by steam autoclaving and Er:YAG laser etching on dentin components.

The simultaneous need for infection-control protocols in sample preparations and for safe laser irradiation parameters prompted this study about the effects of heat produced by both sample sterilization and laser etching on dentin components. The dentin was exposed on 30 bovine incisors, and then divided into two main groups: autoclaved (group A) or thymol treatment (group B). The surface of the dentin was schematically divided into four areas, with each one corresponding to a treatment subgroup. The specimens were either etched with phosphoric acid (control-CG) or irradiated with Er:YAG laser (subgroups: I-80 mJ, II-120 mJ, and III-180 mJ). Elemental distribution maps were done by energy-dispersive X-ray fluorescence (µ-EDXRF) on each treatment area. The dentin surface in depth was exposed and line-scan maps were performed. The B_CG treatment produced the best distribution of calcium (Ca) and phosphorus (P) content throughout the dentin surface. Er:YAG laser etching produced irregular patterns of elemental distribution in the dentin. Laser energies of 120 and 180 mJ produced the highest maximum calcium values. The Er:YAG laser energy of 180 mJ produced a localized increase in Ca and P content on the superficial layer of the dentin (∼ 0-0.10 mm). The autoclaving treatment of samples in experiments is not recommended since it produced damaging effects on dentin components. Er:YAG laser irradiation produced a heterogeneous Ca and P distribution throughout the dentin surface with areas of increased Ca concentration, and this may affect clinically the permeability, solubility, or adhesive characteristics of dental hard tissues with restorative procedures.

Conversion degree of indirect resin composites and effect of thermocycling on their physical properties.

PURPOSE: This study evaluated the degree of conversion (DC) of four indirect resin composites (IRCs) with various compositions processed in different polymerization units and investigated the effect of thermal aging on the flexural strength and Vicker's microhardness. MATERIALS AND METHODS: Specimens were prepared from four IRC materials, namely Gr 1: Resilab (Wilcos); Gr2: Sinfony (3M ESPE); Gr3: VITA VMLC (VITA Zahnfabrik); Gr4: VITA Zeta (VITA Zahnfabrik) using special molds for flexural strength test (N = 80, n = 10 per group) (25 x 2 x 2 mm(3), ISO 4049), for Vicker's microhardness test (N = 80, n = 10 per group) (5 x 4 mm(2)) and for DC (N = 10) using FT-Raman Spectroscopy. For both flexural strength and microhardness tests, half of the specimens were randomly stored in distilled water at 37 degrees C for 24 hours (Groups 1 to 4), and the other half (Groups 5 to 8) were subjected to thermocycling (5000 cycles, 5 to 55 +/- 1 degree C, dwell time: 30 seconds). Flexural strength was measured in a universal testing machine (crosshead speed: 0.8 mm/min). Microhardness test was performed at 50 g. The data were analyzed using one-way and two-way ANOVA and Tukey's test (alpha= 0.05). The correlation between flexural strength and microhardness was evaluated with Pearson's correlation test (alpha= 0.05). RESULTS: A significant effect for the type of IRC and thermocycling was found (p= 0.001, p= 0.001) on the flexural strength results, but thermocycling did not significantly affect the microhardness results (p= 0.078). The interaction factors were significant for both flexural strength and microhardness parameters (p= 0.001 and 0.002, respectively). Thermocycling decreased the flexural strength of the three IRCs tested significantly (p < 0.05), except for VITA Zeta (106.3 +/- 9.1 to 97.2 +/- 14 MPa) (p > 0.05) when compared with nonthermocycled groups. Microhardness results of only Sinfony were significantly affected by thermocycling (25.1 +/- 2.1 to 31 +/- 3.3 Kg/mm(2)). DC values ranged between 63% and 81%, and were not significantly different between the IRCs (p > 0.05). While a positive correlation was found between flexural strength and microhardness without (r = 0.309) and with thermocycling (r = 0.100) for VITA VMLC, negative correlations were found for Resilab under the same conditions (r =-0.190 and -0.305, respectively) (Pearson's correlation coefficient). CONCLUSION: Although all four IRCs presented nonsignificant DC values, flexural strength and microhardness values varied between materials with and without thermocycling.

Comparative study of transdermal drug delivery systems of resveratrol: High efficiency of deformable liposomes.

Trans-resveratrol (3, 5, 4' trihydroxystilbene, RSV) is a natural compound that shows antioxidant, cardioprotective, anti-inflammatory and anticancer properties. The transdermal, painless application of RSV is an attractive option to other administration routes owing to its several advantages like avoiding gastrointestinal problems and first pass metabolism. However, its therapeutic potential is limited by its low solubility and low stability in water and the reduced permeability of stratum corneum. To overcome these inconveniences the encapsulation of this compound in a drug delivery system is proposed here. In order to find the best carrier for transdermal application of RSV various liposomal nanoparticulate carriers like conventional liposomes (L-RSV), deformable liposomes (LD-RSV), ultradeformable liposomes (LUD-RSV) and ethosomes (Etho-RSV) were assayed. Transmission electron microscopic (TEM) and dynamic light scattering (DLS) studies were performed to analyze the surface morphology of these carriers. Structural characterization for these formulations was performed by confocal Raman spectroscopy. The spectroscopic results were analysed in conjunction with calorimetric data to identify the conformational changes and stability of formulations in the different nanoparticles induced by the presence of RSV. Comparison of the results obtained with the different carrier systems (L-RSV, LD-RSV, LUD-RSV and Etho-RSV) revealed that the best RSV carrier was LD-RSV. The increase in the fluidity of the bilayers in the region of the hydrophobic chains of the phospholipid by ethanol probably facilitates the accommodation of the RSV in the bilayer and contributes to the improved encapsulation of RSV without affecting the mobility of this carrier.

Chemical, morphological and thermal effects of 10.6-microm CO2 laser on the inhibition of enamel demineralization.

Studies have shown that enamel can be modified by pulsed CO2 laser to form a more acid-resistant substrate. This study evaluated the effects of a 10.6-microm CO2 laser on enamel surface morphology and chemical composition as well as monitored intrapulpal temperature changes during irradiation. Human teeth were irradiated with fluences of 1.5-11.5 J/cm2, and pulpal thermal as well as chemical and morphological modifications on enamel were assessed. The teeth were submitted to a pH-cycling model, and the mineral loss was determined by means of cross-sectional microhardness. For all irradiated groups, intrapulpal temperature changes were below 3 degrees C. FT-Raman spectroscopy and scanning electron microscopy indicated that fluences as low as 6.0 J/cm2 were sufficient to induce chemical and morphological changes in enamel. Then, for fluences reaching or exceeding 10.0 J/cm2, laser-induced inhibitory effects on demineralization were observed. It was thus concluded that laser energy density in the range of 10.0 and 11.5 J/cm2 could be applied to dental enamel in order to produce chemical and morphological changes and reduce the acid reactivity of enamel without compromising the pulp vitality.

Molecular analysis of Er:YAG laser irradiation on dentin.

The aim of this study was to evaluate by dispersive Raman spectroscopy the mineral and organic components of human dentin before and after laser irradiation and acid etching. The occlusal enamel of six non-carious human third molars was removed providing 6 dentin discs, which were divided in four quadrants each of them receiving a different surface treatment: etching with a 37% phosphoric acid gel (control); irradiation by Er:YAG laser (KaVo Key Laser II) with 80 mJ, 3 Hz, 30 s (group I); 120 mJ, 3 Hz, 30 s (group II); and 180 mJ, 3 Hz, 30 s (group III). The Raman spectra of normal (untreated) and treated dentin were analyzed and the mineral and the organic component were evaluated. Results were submitted to statistical analysis by ANOVA and Tukey's test at 5% significance level. The minerals and organic content were less affected in the control group and group I (p>0.05). Group II presented more reduction in mineral content (p<0.01) whereas in group III the inorganic (p<0.05) and organic (p<0.01) content were more affected. Dispersive Raman spectroscopy provided valid information of dentin chemical constituents with non-chemical sampling preparation.

Influence of mycosporine-like amino acids and gadusol on the rheology and Raman spectroscopy of polymer gels.

BACKGROUND: The amphiphilic nature of polymers allows them to be widely incorporated as carriers in different pharmaceutical applications since they are able to work as vehicles for hydro- or lipo-soluble actives. OBJECTIVE: The aim of this study was to determine the rheological behavior and vibrational spectral variations of two hydrophilic gels prepared with Poloxamer 407 (PO) or Pluronic F-127 (PL) with the addition of the actives mycosporine-like amino acids and gadusol. METHODS: The structures of these polymers in two different concentrations (20% w/w and 27% w/w) were characterized by rheological studies and Raman spectroscopy. RESULTS: Gels prepared with higher polymer concentration showed larger G' (storage modulus) values. The C-C stretch and the CH2 rocking predominated in the gels containing PL (20% w/w) and this correlated with a less viscous behavior. The mixture of the actives induced higher contributions of Raman peaks related to trans conformation of the C-C bonds located in hydrophilic polymer blocks, whereas the same peaks decreased in the sample containing only gadusol. CONCLUSIONS: Larger tensile strength and elastic component were observed upon increasing polymer concentration, thus evidencing polymer-polymer and/or polymer-polymer-actives interactions. The presence of the actives affected the mechanical properties of the polymer gels. Gadusol particularly seems to alter the conformation of the polymer chains by favoring gauche orientations, in parallel with rising viscoelastic parameters. More stretched arrangements of the polymer are probably induced in the presence of larger concentration of actives, due to specific interactions with their hydrophilic groups.

Mineral distribution and CLSM analysis of secondary caries inhibition by fluoride/MDPB-containing adhesive system after cariogenic challenges.

OBJECTIVE: To evaluate the inhibition zone formation (IZ) and mineral distribution along the interface of adhesive systems either containing fluoride and antibacterial primer or not, after chemical and biological artificial caries challenges. METHODS: Forty-eight third molars were used. Artificial caries was developed with S. mutans in a 4mm x 4mm area of occlusal dentin surface. Carious dentin was removed and cavities were restored with Adper Scotchbond Multi-Purpose (SBM) and Clearfil Protect Bond (CPB) (n=24). Samples were submitted to secondary caries development by chemical (C) (acidic gel) or biological (B) (S. mutans culture) methods for 5 days. Four groups were tested (n=12): (1) SC (SBM+C); (2) SB (SBM+B); (3) CC (CPB+C); (4) CB (CPB+B). The IZ and outer lesion (OL) formations were analyzed by confocal laser scanning microscopy (CLSM). The distribution of calcium (Ca) and phosphorus (P) content along the interface was analyzed by micro X-ray fluorescence spectrometer by energy-dispersive (microEDX). RESULTS: The frequency of IZ formation and mean values of IZ thickness differed among the groups. The CC group presented the lowest OL depth. microEDX analysis showed that CPB had the highest mineral loss by the biological method, but the lowest mineral loss by the chemical method. SC and SB groups showed intermediate values of mineral loss. CONCLUSION: The mineral loss along the dentin/restoration interface was affected by the artificial caries method, and hybrid layer formation by adhesive systems used. The adhesive system containing fluoride and antibacterial primer did not prevent secondary caries formation.

Molecular analysis of Er: YAG laser irradiation on dentin

O objetivo deste estudo foi avaliar por espectroscopia Raman, os componentes mineral e orgânico da dentina humana antes e após o condicionamento ácido e a irradiação com laser de Er:YAG. Seis discos de dentina foram obtidos de 6 terceiros molares humanos hígidos após remoção da superfície oclusal. Cada disco foi dividido em quatro regiões (quadrantes) de tratamento: condicionamento com ácido fosfórico a 37% (controle), irradiação com laser de Er:YAG (KaVo Key Laser II) com 80 mJ, 3 Hz, 30 s (grupo I); 120 mJ, 3 Hz, 30 s (grupo II) e 180 mJ, 3 Hz, 30 s (grupo III). Os espectros Raman da dentina normal e tratada foram analisados e os componentes mineral e orgânico foram avaliados. Os resultados foram submetidos a análise estatística pela ANOVA e teste de Tukey com intervalo de confiança de 95%. O conteúdo mineral e orgânico foi menos afetado nos grupos controle e I (p>0,05). O grupo II apresentou maior redução no conteúdo mineral (p<0,01) enquanto que a irradiação com laser Er:YAG 180 mJ (grupo III) reduziu mais o conteúdo inorgânico (p<0,05) e orgânico (p<0,01). A espectroscopia Raman forneceu informações dos conteúdos químicos da dentina sem preparação química dos espécimes.