Topical dexpanthenol effects on physiological parameters of the stratum corneum by Confocal Raman Microspectroscopy.

BACKGROUND: Topical use of dexpanthenol presents well-established moisturizing properties and maintenance and repair of the skin barrier function, however, its exact action mechanisms are not completely elucidated. In this context, Confocal Raman Microspectroscopy is an optical method that enables non-invasive and non-destructive in vivo analysis with the sensitive acquisition of molecular changes in different skin layers. Herein, the aim was to evaluate the effects of topical dexpanthenol on the components and physiological parameters of the stratum corneum (SC). MATERIALS AND METHODS: Ten healthy female subjects underwent skin evaluation by means of a Confocal Raman Spectrometer Skin Analyzer 3510. Spectral data were obtained from the skin of the anterior forearm region, before and 2 h after applying a cosmetic formulation containing or not containing 5% dexpanthenol. RESULTS: Semiquantitative analysis of the natural moisturizing factor showed a significant decrease in content after 2 h of topical dexpanthenol application, while the analysis of the lamellar organization of intercellular lipids and the secondary structure of keratin showed a significant increase in hexagonal organization of lipids at the first half of the SC and a significant increase in ß-pleated sheet conformation of keratin. CONCLUSION: Effects of topical dexpanthenol on SC suggest a contribution in increasing fluidity of both lipidic and protein components of the SC and are compatible with dexpanthenol activity in maintaining adequate physiological conditions and preventing transepidermal water loss. This study also contributes to the elucidation of action mechanisms and other concurrent biochemical processes.

Correlation between ankle-brachial index and thermography measurements in patients with peripheral arterial disease.

OBJECTIVES: To correlate the ankle-brachial index and photographic thermography findings in patients with peripheral arterial disease. METHODS: Photographic thermography was performed at the foot level, and ankle-brachial index was measured in 72 lower limbs of 53 patients with peripheral arterial disease who were divided into calcified artery, patients with an ankle-brachial index greater than 1.4; and non-calcified artery classified as asymptomatic, mild, moderate, and severe on the basis of peripheral arterial disease severity. Fisher's exact test was used for categorical data, and Wilcoxon test was used for numerical data. RESULTS: Spearman's correlation analysis showed a strong correlation (R = 0.7) between the ankle-brachial index and the mean plantar temperature in patients without lower limb artery calcification. Linear regression yielded the predictor equation Y = 3.296 × X + 29.75, wherein ankle-brachial index (X) can be predicted on the basis of temperature values. Spearman's correlation test showed no significance (p = 0.2174) in patients with arterial calcification. Kruskal-Wallis test with post hoc analysis using Dunn's test for multiple comparisons showed that the mean plantar temperature was lower in patients with arterial calcification. CONCLUSION: Photographic thermography findings show a strong correlation with ankle-brachial index in patients with non-calcified arteries.

Short-term and long-term effects of osteoporosis on incisor teeth and femoral bones evaluated by Raman spectroscopy and energy dispersive X-ray analysis in ovariectomized rats.

There are few published data on the relationship between loss of bone mass due to osteoporosis and poor tooth quality. This study analyzed the effects of osteoporosis on incisor teeth and femoral bones using optical techniques in rats. Twenty female Wistar rats aged 6 months (n = 20) were randomized into two groups: control group, non-ovariectomized rats (n = 10); ovariectomy group, ovariectomized rats to induce osteoporosis (n = 10). Each group was subdivided randomly into two groups containing five rats each as follows. Control group 1: non-ovariectomized rats euthanized at the age of 9 or 3 months post-ovariectomy (n = 5); Control group 2: non-ovariectomized rats euthanized at the age of 1 year or 6 months post-ovariectomy (n = 5); ovariectomy group 1: ovariectomized rats euthanized at the age of 9 months or 3 months post-ovariectomy (n = 5); ovariectomy group 2: ovariectomized rats euthanized at the age of 1 year or 6 months post-ovariectomy (n = 5). The incisor teeth and femoral bones of Wistar rats were removed to perform Raman spectroscopy using an excitation laser at 785 nm. In addition, an energy-dispersive X-ray spectrometer system was used to evaluate calcium (Ca) and phosphorus (P). The main findings included significant changes (p < 0.05) for phosphate and carbonate band areas for both incisor teeth and femur bones. In addition, there was significant negative correlation between the P concentration and phosphate/carbonate ratio (lower P content-larger ratio, p < 0.05) for incisor teeth and femoral bones. The proline and CH2 wag band areas were significantly reduced only for the incisor teeth (p < 0.05). Therefore, Raman spectroscopy assessed the compositional, physicochemical and structural changes in hard tissue. The current study also pointed out the possible action mechanisms of these changes, bone fracture risk and dental fragility. It is important to emphasize that poor dental quality may also occur due to osteoporosis.

In vivo study of dermal collagen of striae distensae by confocal Raman spectroscopy.

This research work mainly deals with studying qualitatively the changes in the dermal collagen of two forms of striae distensae (SD) namely striae rubrae (SR) and striae albae (SA) when compared to normal skin (NS) using confocal Raman spectroscopy. The methodology includes an in vivo human skin study for the comparison of confocal Raman spectra of dermis region of SR, SA, and NS by supervised multivariate analysis using partial least squares discriminant analysis (PLS-DA) to determine qualitatively the changes in dermal collagen. These groups are further analyzed for the extent of hydration of dermal collagen by studying the changes in the water content bound to it. PLS-DA score plot showed good separation of the confocal Raman spectra of dermis region into SR, SA, and NS data groups. Further analysis using loading plot and S-plot indicated the participation of various components of dermal collagen in the separation of these groups. Bound water content analysis showed that the extent of hydration of collagen is more in SD when compared to NS. Based on the results obtained, this study confirms the active involvement of dermal collagen in the formation of SD. It also emphasizes the need to study quantitatively the role of these various biochemical changes in the dermal collagen responsible for the variance between SR, SA, and NS.

In Vivo Human Skin Penetration Study of Sunscreens by Confocal Raman Spectroscopy.

This research work mainly deals with the application of confocal Raman spectroscopic technique to study in vivo human skin penetration of sunscreen products, as there are a lot of controversies associated with their skin penetration. Healthy human volunteers were tested for penetration of two commercial sunscreen products into their volar forearm skin for a period of 2 h. Measurements were taken before and after application of these sunscreen products. All the confocal Raman spectra were pre-processed and then subjected to multivariate two-dimensional principal component analysis and classical least squares analysis to determine the skin penetration of these sunscreens in comparison to the "sunscreen product spectrum" which was considered as the control. Score plots of principal component analysis of confocal Raman spectra indicated clear separation between the spectra before and after application of sunscreen products. Loading plots showed the maximum differences in the spectral region from 1590 to 1626 cm-1 where the characteristic peak of the pure sunscreen products was observed. Classical least squares analysis has shown a significant penetration to a depth of 10 µm in the volar forearm skin of healthy human volunteers for both these sunscreen products. The results confirm that the penetration of these tested sunscreen products was restricted to stratum corneum and also prove that confocal Raman spectroscopy is a simple, fast, nondestructive, and noninvasive semi-quantitative analytical technique for these studies.

In Vivo Determination of Moisturizers Efficacy on Human Skin Hydration by Confocal Raman Spectroscopy.

This research work deals with in vivo testing of the efficacy of commercial moisturizer products on the hydration of human skin, as there are various in vitro and ex vivo studies questioning their activity. Confocal Raman spectroscopy was used for this purpose of assessing the efficacy of moisturizers on skin hydration mainly owing to its simple, non-invasive, non-destructive, timesaving, and cost-effective nature. Water content and natural moisturizing factor (NMF) of stratum corneum were analyzed and compared using this method at high wavenumber (2500-4000 cm-1) and fingerprint (400-1800 cm-1) spectral regions, respectively, as these two parameters are correlated to skin hydration. Four commercial moisturizer products of different brands were tested on volar forearm region of healthy human female volunteers. This study was conducted for a period of 30 days with 0, 7, and 30 days as time points of analysis. The results of this study clearly indicate that not all the moisturizer products hydrate the skin to the expected levels, and this extent of skin hydration varies with duration of application of these products.

In vitro effects of hydrogen peroxide combined with different activators for the in-office bleaching technique on enamel.

OBJECTIVE: The aim of this study was to evaluate the alteration of human enamel bleached with high concentrations of hydrogen peroxide associated with different activators. MATERIALS AND METHODS: Fifty enamel/dentin blocks (4 × 4 mm) were obtained from human third molars and randomized divided according to the bleaching procedure (n = 10): G1 = 35% hydrogen peroxide (HP - Whiteness HP Maxx); G2 = HP + Halogen lamp (HL); G3 = HP + 7% sodium bicarbonate (SB); G4 = HP + 20% sodium hydroxide (SH); and G5 = 38% hydrogen peroxide (OXB - Opalescence Xtra Boost). The bleaching treatments were performed in three sessions with a 7-day interval between them. The enamel content, before (baseline) and after bleaching, was determined using an FT-Raman spectrometer and was based on the concentration of phosphate, carbonate, and organic matrix. Statistical analysis was performed using two-way ANOVA for repeated measures and Tukey's test. RESULTS: The results showed no significant differences between time of analysis (p = 0.5175) for most treatments and peak areas analyzed; and among bleaching treatments (p = 0.4184). The comparisons during and after bleaching revealed a significant difference in the HP group for the peak areas of carbonate and organic matrix, and for the organic matrix in OXB and HP+SH groups. Tukey's analysis determined that the difference, peak areas, and the interaction among treatment, time and peak was statistically significant (p < 0.05). CONCLUSION: The association of activators with hydrogen peroxide was effective in the alteration of enamel, mainly with regards to the organic matrix.

Effects of low-power LED and therapeutic ultrasound in the tissue healing and inflammation in a tendinitis experimental model in rats.

This work evaluated the anti-inflammatory response of low-power light-emitting diode (LED) and ultrasound (US) therapies and the quality and rapidness of tendon repair in an experimental model of tendinitis, employing histomorphometry and Raman spectroscopy. Tendinitis was induced by collagenase into the right tendon of 35 male Wistar rats with an average weight of 230 g. The animals were randomly separated into seven groups of five animals each: tendinitis without treatment-control (TD7 and TD14, where 1 and 2 indicated sacrifice on the 7th and 14th day, respectively), tendinitis submitted to US therapy (US7 and US14) and tendinitis submitted to LED therapy (LED7 and LED14). Contralateral tendons of the TD group at the 14th day were used as the healthy group (H). US treatment was applied in pulsed mode at 10 %, 1 MHz frequency, 0.5 W/cm(2), 120 s. LED therapy parameters were 4 J/cm(2), 120 s, daily dose at the same time and same point. Sacrifice was performed on the 7th or 14th day. Histomorphometric analysis showed lower number of fibroblasts on the 14th day of therapy for the US-treated group, compared to the TD and LED, indicating lower tissue inflammation. Raman showed that the LED group had an increase in the amount of collagen I and III from the 7th to the 14th day, which would indicate more organized fibers and a better quality of the healing, and US showed lower collagen I synthesis in the 14th day compared to H, indicating a lower tissue reorganization.

FT-Raman spectroscopy study of organic matrix degradation in nanofilled resin composite.

This in vitro study evaluated the effect of light curing unit (LCU) type, mouthwashes, and soft drink on chemical degradation of a nanofilled resin composite. Samples (80) were divided into eight groups: halogen LCU, HS--saliva (control); HPT--Pepsi Twist®; HLC--Listerine®; HCP--Colgate Plax®; LED LCU, LS--saliva (control); LPT--Pepsi Twist®; LLC--Listerine®; LCP--Colgate Plax®. The degree of conversion analysis and the measure of the peak area at 2,930 cm-1 (organic matrix) of resin composite were done by Fourier-transform Raman spectroscopy (baseline, after 7 and 14 days). The data were subjected to multifactor analysis of variance (ANOVA) at a 95% confidence followed by Tukey's HSD post-hoc test. The DC ranged from 58.0% (Halogen) to 59.3% (LED) without significance. Differences in the peak area between LCUs were found after 7 days of storage in S and PT. A marked increase in the peak intensity of HLC and LLC groups was found. The soft-start light-activation may influence the chemical degradation of organic matrix in resin composite. Ethanol contained in Listerine® Cool Mint mouthwash had the most significant degradation effect. Raman spectroscopy is shown to be a useful tool to investigate resin composite degradation.

Identification of biomarkers in diabetic nails by Raman spectroscopy.

BACKGROUND: The growth of Diabetes Mellitus (DM) is a serious public health issue which is more prevalent in developing countries. The main problems related to DM are the gradual changes in the structural and functional integrity of tissues caused by hyperglycemia, which calls for early diagnosis and periodic monitoring exams. Recent studies suggest that the quality of the nail plate has great potential to assess the secondary complications of DM. Hence, this study aimed to determine the biochemical characteristics of the nails of individuals with DM2 by Raman confocal spectroscopy (CRS). METHODS: We collected fragments from the distal region of the fingernails of 30 healthy volunteers and 30 volunteers with DM2. The samples were analyzed by CRS (Xplora - Horiba) coupled to a 785 nm laser. RESULTS: Alterations in different biochemical components, such as proteins, lipids, amino acids, and final agents of advanced glycation, and alterations in the disulfide bridges, which are important in stabilizing keratin in nails were identified. CONCLUSION: The spectral signatures and new DM2 markers in nails were identified. Therefore, the possibility of acquiring biochemical information by evaluating the nails of diabetics, a simple and easily acquired material associated with the CRS technique, may allow health complications to be detected quickly.

Scanning electron microscopy and roughness study of dental composite degradation.

Our aim was to test the hypothesis that the use of mouthwashes, consumption of soft drinks, as well as the type of light curing unit (LCU), would change the surface roughness (Ra) and morphology of a nanofilled composite resin (Z350® 3M ESPE). Samples (80) were divided into eight groups: Halogen LCU, group 1, saliva (control); group 2, Pepsi Twist®; group 3, Listerine®; group 4, Colgate Plax®; LED LCU, group 5, saliva; group 6, Pepsi Twist®; group 7, Listerine®; group 8, Colgate Plax®. Ra values were measured at baseline, and after 7 and 14 days. One specimen of each group was prepared for scanning electron microscopy analysis after 14 days. The data were subjected to multifactor analysis of variance at a 95% confidence followed by Tukey's honestly significant difference post-hoc test. All the treatments resulted in morphological changes in composite resin surface, and the most significant change was in Pepsi Twist® groups. The samples of G6 had the greatest increase in Ra. The immersion of nanofilled resin in mouthwashes with alcohol and soft drink increases the surface roughness. Polymerization by halogen LCU (reduced light intensity) associated with alcohol contained mouthwash resulted in significant roughness on the composite.

Micro energy-dispersive X-ray fluoresence mapping of enamel and dental materials after chemical erosion.

Energy-dispersive X-ray fluorescence was employed to test the hypothesis that beverage consumption or mouthwash utilization will change the chemical properties of dental materials and enamel mineral content. Bovine enamel samples (n = 45) each received two cavity preparations (n = 90), each pair filled with one of three dental materials (R: nanofilled composite resin; GIC: glass-ionomer cement; RMGIC: resin-modified GIC). Furthermore, they were treated with three different solutions (S: saliva; E: erosion/Pepsi Twist®; or EM: erosion+mouthwash/Colgate Plax®). It was found that mineral loss in enamel was greater in GICE samples than in RE > RMGICE > RMGICEM > REM > GICEM. An increased percentage of Zr was found in REM indicating organic matrix degradation. Dental materials tested (R, GIC, and RMGIC) were not able to protect adjacent enamel from acid erosion by the soft drink tested. The use of mouthwash promoted protection of enamel after erosion by the soft drink. To avoid chemical dissolution by mouthwashes, protection by resin composites with surface sealants is recommended.

Influence of creatine supplementation on bone quality in the ovariectomized rat model: an FT-Raman spectroscopy study.

The influence of creatine (Cr) supplementation on cortical and trabecular bone from ovariectomized rats was studied using FT-Raman spectroscopy. The intensity of organic-phase Raman bands was compared to mineral phase ones. Twenty-one female Wistar rats aged 3 months were divided into three groups (n = 7 per group): ovariectomized (OVX), ovariectomized treated with creatine (CRE) and sham-operated (SHAM) groups. Creatine supplementation (300 mg kg(-1) day(-1)) was provided for 8 weeks, starting 12 weeks after ovariectomy. FT-Raman spectroscopy was performed on the right medial femoral mid-shaft (cortical bone) and third lumbar vertebral body (trabecular bone). The integrated intensities of mineral phase (phosphate and carbonate bands at 959 and 1,071 cm(-1), respectively) and organic phase (amide I band at 1,665 cm(-1)) Raman bands were analyzed. The mineral-to-matrix (phosphate/amide I), carbonate-to-phosphate, and carbonate-to-amide I ratios were analyzed to assess bone quality. The phosphate content on trabecular bone was higher in the CRE group than the OVX group (p < 0.05). No significant changes in mineral or organic phases on cortical bone were observed. A radiographic assessment of bone density was encouraging as the same findings were showed by Raman intensity of phosphate from cortical (r(2) = 0.8037) and trabecular bones (r(2) = 0.915). Severe ovariectomy-induced bone loss was confirmed by FT-Raman spectroscopy. The results suggest that the chemical composition of trabecular bone tissue may be positively influenced by Cr supplementation after ovariectomy.

Effects of Er:YAG laser irradiation and manipulation treatments on dentin components, part 2: energy-dispersive X-ray fluorescence spectrometry study.

The effects of laser etching, decontamination, and storage treatments on dentin components were studied by energy-dispersive X-ray fluorescence spectrometry (EDXRF). Thirty bovine incisors were prepared to expose the dentin surface and then divided into two main groups based upon the decontamination process and storage procedure: autoclaved (group A, n=15) or stored in aqueous thymol solution (group B, n=15). The surfaces of the dentin slices were schematically divided into four areas, with each one corresponding to a treatment subgroup. The specimens were either etched with phosphoric acid (control subgroup) or irradiated with erbium-doped yttrium-aluminum-garnet (Er:YAG) laser (subgroups: I-80 mJ, II-120 mJ, and III-180 mJ). Samples were analyzed by micro-EDXRF, yielding three spectra for each area (before and after treatment). Surface mappings covering an area of 80x60 points with steps of 20 mum were also performed on selected specimens. The amount of Ca and P in group A specimens decreased significantly (P<0.05) after the acid etching and the CaP ratio increased (P<0.001). Er:YAG laser-etching using lower laser energies did not produce significant changes in dentin components. The mapping data support the hypothesis that acid etching on dentin produced a more chemically homogeneous surface and thus a more favorable surface for the diffusion of adhesive monomers.

Effects of Er:YAG laser irradiation and manipulation treatments on dentin components, part 1: Fourier transform-Raman study.

The effects of laser etching, decontamination, and storage treatments on dentin components were studied using Fourier transform (FT)-Raman spectroscopy. Thirty bovine incisors were prepared to expose the dentin surface and then divided in two main groups based upon the decontamination process and storage procedure: autoclaved (group A, n=15) or stored in thymol aqueous solution (group B, n=15). The surfaces of the dentin slices were schematically divided into four areas, with each one corresponding to a treatment subgroup. The specimens were either etched with phosphoric acid (control subgroup) or irradiated with erbium-doped yttrium-aluminum-garnet (Er:YAG) laser (subgroups: I-80 mJ, II-120 mJ, and III-180 mJ, and total energy of 12 J). Samples were analyzed by FT-Raman spectroscopy; we collected three spectra for each area (before and after treatment). The integrated areas of five Raman peaks were calculated to yield average spectra. The areas of the peaks associated with phosphate content (P<0.001), type I collagen, and organic C-H bonds (P<0.05) were reduced significantly in group A (control). Analyses of samples irradiated with reduced laser energies did not show significant changes in the dentin components. These results suggest that thymol storage treatment is advised for in vitro study; furthermore, 12 J of Er:YAG laser energy does not affect dentin components.

Effects of acidic primer/adhesives on primary and permanent dentin.

PURPOSE: To evaluate the quality of primary and permanent dentin by Fourier transformed Raman spectroscopy (FT-Raman), and scanning electron microscopy/energy-dispersive spectroscopy (SEM/EDS). METHODS: The middle dentin of crowns was reached by carbide bur abrading providing a uniform smear layer. Self-etching primers were applied in order to simulate the etching of self-etching adhesive systems. The groups were (n = 6): G1 (primary dentin smear layer); G2 (primary dentin etched by primer of Clearfil Protect Bond); G3 (primary dentin etched by Adper Prompt); G4 (permanent dentin smear layer); G5 (permanent dentin etched by primer of Clearfil Protect Bond); G6 (permanent dentin etched by Adper Prompt). SEM/EDS were made in order to obtain additional elemental data to complement FT-Raman. FT-Raman data were submitted to cluster analysis. RESULTS: Overall, FT-Raman showed differences between primary and permanent dentin concerning organic content, but not for inorganic content. FT-Raman showed differences in the organic content between primary and permanent dentin after self-etching primer use. HEMA usage caused molecular changes in the organic content, while phosphoric acidic ester caused molecular changes in the inorganic content of primary and permanent dentin. The SEM/EDS identified C, O, P, and Ca, which could not replace ions to change mineral molecular arrangement. Both organic and mineral content arrangements were similar after self-etching primers action. The organic content of dentin was modified by both Clearfil PB primer and Adper Prompt for primary dentin but only by Clearfil PB for permanent dentin. The inorganic content of primary middle dentin was similar to the inorganic content of permanent middle dentin, even when Adper Prompt is used.

Diagnosis of degenerative lesions of supraspinatus rotator cuff tendons by Fourier transform-Raman spectroscopy.

The Fourier transform (FT)-Raman spectroscopy technique is used to assess the biochemical alterations that occur in the degenerative process of the rotator cuff supraspinatus tendon. The main alterations observed occur in the glycine, proline, hydroxyproline, cysteine, cistine, phenylalanine, tyrosine, collagen I and III, nucleic acid, lipids, glycosaminoglycans, and metalloproteinases bands. An increasing intensity for these bands is found in degenerated tendons, a finding well correlated with hyaline state and cellular activity. Statistical analysis (principal components analysis and clustering) shows a clear separation of the spectra into nonhyalinized and hyalinized clusters, which enables the construction of a binary diagnosis model based on logistic regression. Best diagnosis provided a sensitivity of 66.0% and a specificity of 74.7% with 79.6% concordant pairs. The discriminating power of the diagnostic test is assessed by computing the area under the receiving-operator characteristic curve (AUC), which indicates good accuracy (AUC=0.81). In principle, these results indicate that Raman spectroscopy can be used as an auxiliary aid to improve shoulder tendon surgery quality by guiding anchoring onto more healthy (nonhyaline) pieces of tendons. This should contribute to a decrease in the current high rerupture rate (13 to 68%) for this procedure.

Role of cervicitis in the Raman-based optical diagnosis of cervical intraepithelial neoplasia.

The Raman-based optical diagnosis of normal cervix, inflammative cervix (cervicitis), and cervical intraepithelial neoplasia was investigated on samples of 63 patients. The main alterations were found in the 857 cm(-1) (CCH deformation aromatic); 925 cm(-1) (C-C stretching); approximately 1247 cm(-1) (CN stretch, NH bending of Amide III); 1370 cm(-1) (CH2 bending); and 1525 cm(-1) (C=CC=N stretching) vibrational bands in accordance with previously reported in the literature comparing normal and malignant cervical tissue. The statistical analysis (principal components analysis, clustering, and logistic regression models) applied to the spectral data indicated that the full discrimination among normal and neoplastic tissues of cervix by Raman optical biopsy is seriously affected by the presence of inflammatory infiltrates, which increases the false-positive rate. This fact is specially relevant once cervicitis is a very common state (noncancerous) of the cervix of sexually active woman. The results suggest that, for the correct Raman-based diagnosis of normal cervix from cervical intraepithelial neoplasia, it is necessary to use an auxiliary way to discriminate the contribution from the inflammatory infiltrates.

NaOCl effects on primary and permanent pulp chamber dentin.

OBJECTIVES: The dentin quality of primary and permanent pulp chamber was inspected by Fourier-transformed Raman spectroscopy (FT-Raman) and scanning electron microscopy (SEM). Fragments of pulp chamber dentin were obtained from 20 human molar crowns (primary and permanent). METHODS: The fragments were assigned to 8 groups (n=5)-Primary teeth: G1, pulp chamber dentin; G2, pulp chamber dentin irrigated with NaOCl 1% (30 min); G3, pulp chamber dentin irrigated with NaOCl 1% (30 min) and etched by 35% phosphoric acid; G4, pulp chamber dentin etched by 35% phosphoric acid. Permanent teeth: G5, pulp chamber dentin; G6, pulp chamber dentin irrigated with NaOCl 1% (30 min); G7, pulp chamber dentin irrigated with NaOCl 1% (30 min) and etched by 35% phosphoric acid; G8, pulp chamber dentin etched by 35% phosphoric acid. The spectra were subjected to the Cluster analysis. The SEM images were scored. RESULTS: Inorganic content: There was a difference between primary and permanent dentin. The groups treated with NaOCl were statistically similar between them, but differed from the groups not treated. Organic content: There was no difference between primary and permanent dentin. The groups became similar after NaOCl and phosphoric acid treatments. The microscopic images showed the presence of calcospherites on permanent dentin and their absence on primary dentin. CONCLUSIONS: The NaOCl changed the inorganic content in both dentitions; regardless of the following phosphoric acid etching. However, the chemical changes caused by NaOCl were not detected by SEM when it was followed by etching.

In vitro effects of alcohol-containing mouthwashes on human enamel and restorative materials.

The objective of this study was to evaluate the in vitro effects, including surface morphological characteristics and chemical elemental properties, of different mouthwash formulations on enamel and dental restorative materials, simulating up to 6 months of daily use. Human enamel samples, hydroxyapatite, composite resin, and ceramic surfaces were exposed to 3 different mouthwashes according to label directions - Listerine® Cool Mint®, Listerine® Total Care, and Listerine® Whitening - versus control (hydroalcohol solution) to simulate daily use for up to 6 months. The samples were analyzed using scanning electron microscopy (SEM), infrared spectrophotometry (µ-Fourier transform infrared microscopy), energy-dispersive X-ray (EDX) spectroscopy, and color analysis before and after exposure. No relevant changes were observed in the morphological characteristics of the surfaces using SEM techniques. The physical and chemical aspects of the enamel surfaces were evaluated using mid-infrared spectroscopy, and EDX fluorescence was used to evaluate the elemental aspects of each surface. There was no variation in the relative concentrations of calcium and phosphorus in enamel, silicon and barium in composite resin, and silicon and aluminum in the ceramic material before and after treatment. No relevant changes were detected in the biochemical and color properties of any specimen, except with Listerine® Whitening mouthwash, which demonstrated a whitening effect on enamel surfaces. Long-term exposure to low pH, alcohol-containing, and peroxide-containing mouthwash formulations caused no ultra-structural or chemical elemental changes in human enamel or dental restorative materials in vitro.