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

Abstract 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.

Molecular and morphological surface analysis: effect of filling pastes and cleaning agents on root dentin

Abstract The quality of the dentin root is the most important factor for restoration resin sealing and drives the outcome of endodontic treatment. Objective This study evaluated the effect of different filling pastes and cleaning agents on the root dentin of primary teeth using Fourier-transformed Raman spectroscopy (FT-Raman), micro energy-dispersive X-ray fluorescence (µ-EDXRF) and scanning electron microscopic (SEM) analysis. Material and Methods Eighty roots of primary teeth were endodontically prepared and distributed into 4 groups and filled according to the following filling pastes: Control-no filling (CP), Calen®+zinc oxide (CZ), Calcipex II® (CII), Vitapex® (V). After seven days, filling paste groups were distributed to 4 subgroups according to cleaning agents (n=5): Control-no cleaning (C), Ethanol (E), Tergenform® (T), 35% Phosphoric acid (PA). Then, the roots were sectioned and the dentin root sections were internally evaluated by FT-Raman, µ-EDXRF and SEM. Data was submitted to two-way ANOVA and Tukey tests (α=0.05). Results Regarding filling pastes, there was no significant difference in organic content. CP provided the lowest calcium values and, calcium/phosphoric ratio (Ca/P), and the highest phosphoric values. For cleaning agents there was no difference in organic content when compared to the C; however, T showed significantly higher calcium and Ca/P than PA. All groups showed similar results for phosphorus. The dentin smear layer was present after use of the cleaning agents, except PA. Conclusion The filling pastes changed the inorganic content, however they did not change the organic content. Cleaning agents did not alter the inorganic and organic content. PA cleaned and opened dentin tubules.

Estudo de linfonodos por espectroscopia Raman confocal / Lymph node study by confocal Raman spectroscopy

Dentre os órgãos linfoides encontram-se os linfonodos. Também conhecidos como nódulos linfáticos, os quais são considerados como filtros da linfa por serem ricos em linfócitos e outras células de defesa. Entre os fatores que contribuem para o seu comprometimento temos a invasão de sua estrutura por células neoplásicas. A citologia constitui o exame padrão ouro no diagnóstico sendo, a PAAF a técnica mais utilizada para a obtenção do material a ser analisado. Entretanto, alguns estudos questionam a sua utilização, devido à perda da arquitetura celular. A espectroscopia Raman confocal por meio da sua capacidade em utilizar as diferenças bioquímicas de tecidos e células vem se destacando cada vez mais neste tipo de diagnóstico. O estudo teve por objetivo identificar as principais características bioquímicas em linfonodos normais, comprometidos e lesão cervical utilizando a espectroscopia Raman confocal. Foram analisadas 12 amostras de linfonodos normais e alterados de pacientes com indicação cirúrgica para linfadenectomia pela espectroscopia seguida pela análise estatística de componentes principais (PCA) e análise de discriminante linear (LDA). Após as análises observou-se que a técnica utilizada conseguiu discriminar os tecidos com valores de especificidade, sensibilidade e acurácia de 83%. Portanto, a espectroscopia Raman confocal se mostrou uma importante ferramenta na caracterização de linfonodos podendo futuramente auxiliar no diagnóstico de lesões linfonodais.

Among the lymphoid organs there are lymph nodes,, which are considered as lymph filters because they are rich in lymphocytes and other defense cells. Among the factors that contribute to their impairment we note the invasion of its structure by neoplastic cells. Cytology is the gold-standard diagnostic test, and FNA is the technique most used to obtain the material to be analyzed. However, some studies question its use, due to the loss of cellular architecture. Confocal Raman spectroscopy, through its ability to utilize biochemical differences in tissues and cells, has been increasingly prominent in this type of diagnosis. The study aimed to identify the main biochemical characteristics in normal and compromised lymph nodes and cervical lesion using confocal Raman spectroscopy. We analyzed 12 normal and altered lymph node samples from patients with medical indications for a lymphadenectomy by spectroscopy, followed by statistical analysis of the main components (PCA) and linear discriminant analysis (LDA). After the analysis, it was observed that this technique was able to discriminate the tissues with values of specificity, sensitivity and accuracy of up to 83%. Therefore, confocal Raman spectroscopy has been shown to be an important tool in the characterization of lymph nodes, and may help in the diagnosis of lymph node lesions in the future.

Biochemical imaging of normal, adenoma, and colorectal adenocarcinoma tissues by Fourier transform infrared spectroscopy (FTIR) and morphological correlation by histopathological analysis: preliminary results

Introduction The colorectal cancer is a major health problem worldwide. Histology is considered the gold standard for differential diagnosis. However, it depends on the observer's experience, which can lead to discrepancies and poor results. Spectroscopic imaging by Fourier transform infrared (FTIR) is a technique that may be able to improve the diagnosis, because it is based on biochemical differences of the structural constituents of tissue. Therefore, the main goal of this study was to explore the use of FTIR imaging technique in normal colon tissue, colorectal adenoma, and adenocarcinoma in order to correlate their morphological structures with their biochemical imaging. Methods Samples were collected from normal (n = 4), adenoma (n = 4), and adenocarcinoma human colorectal tissue (n = 4) from patients undergoing colonoscopy or surgical resection of colon lesions. The samples were sectioned with a cryostat in sequential sections; the first slice was placed on CaF2 slide and the second slice was placed on glass slide for histological analysis (HE staining). The cluster analyses were performed by the software Cytospec (1.4.02)®. Results In normal samples, biochemical analysis classified six different structures, namely the lamina propria of mucous glands (epithelial cells and goblet cells), central lumen of the gland, mucin, and conjunctive tissue. In samples with adenoma and adenocarcinoma, altered regions could also be identified with high sensitivity and specificity. Conclusion The results of this study demonstrate the potential and viability of using infrared spectroscopy to identify and classify colorectal tissues.

Evaluation of inorganic and organic bone components after application of an apatite-coated Al2O3 implants as scaffolds for bone repair

The aim of this work was to study the influence of uncoated and apatite-coated Al2O3 implants on bone regeneration after 30 days of surgery in New Zealand white rabbits. Twelve samples of Al2O3 implants were prepared and half of them (n = 6) were apatite-coated by the modified biomimetic method. Three experimental groups were tested as Group C - control, surgery procedure without cerami c implant, Group Ce - uncoated Al 2O3 implants (n = 6) and Group CeHA - apatite-coated Al2O3 implants (n = 6). The mineralization of the Al2O3 implants was analyzed in bone fragments using FT-Raman spectroscopy. Raman peaks at 959 cm-1 and 2940 cm-1 evaluated the inorganic and organic bone content, respectively. In vivo citotoxicity was analyzed using micronucleus test. Inorganic and organic content were higher in CeHA samples than in Ce and C (CeHA > Ce > C). FT-Raman spectroscopy showed that the higher the deposition of the organic matrix, more mineralization occurred. The micronucleus test showed that the uncoated and apatite-coated Al2O3 implants were non-cytotoxic and safe to in vivo applications.

Confocal Raman spectroscopy: determination of natural moisturizing factor profile related to skin hydration

INTRODUCTION: Skin health and skin care to reduce the effects of aging are the main interests of many researchers. The skin is very important because it protects the body from various effects of the external environment, and studies of the largest organ of the human body have been conducted since antiquity. In skin, aging effects are severe enough to promote changes in cell structure and biochemical composition. In this study, we quantitatively analyzed the water content and natural moisturizing factor of human facial skin in vivo and in real time by confocal Raman spectroscopy. This non-invasive technique is capable of providing detailed information on the biochemical composition at different depth profiles in the skin. METHODS: We studied 10 volunteers, phototype II (40 and 50 years old), using a confocal Raman system to examine the skin surface down to 25 µm. Raman spectra were obtained before product use (T0), and after 30 days of continuous use of cosmetics (T30). RESULTS: The results show a significant increase of 6.4% in water content in the surface layer of the facial skin after the cosmetic use. The amounts of natural moisturizing factor (NMF) compounds were also increased. Urocanic acid underwent a greater change in relation to carboxylic acid pyrrolidone, with a 38.5% increase in the stratum corneum. CONCLUSION: Confocal Raman spectroscopy identified changes in the biochemical composition of the superficial layers of the epidermis, which suggests the anti-aging efficacy of the formulation.

FT-Raman spectroscopic study of skin wound healing in diabetic rats treated with Cenostigma macrophyllum Tul

INTRODUCTION: Patients with diabetes mellitus exhibit a delay in the lesion repair process. The active components of Cenostigma macrophyllum may represent a viable alternative to facilitate the recovery of these lesions. The aim of this study was to evaluate the effects of emulsion oil-water Cenostigma macrophyllum in the repair process of lesions in rats with induced diabetes. METHODS: 63 male rats (Wistar, 200-250 g body weight, 30-40 days old) were distributed into the following groups: control (C), diabetic (D) and diabetic treated with Cenostigma macrophyllum (P), subdivided based on the experimental times, days 7, 14 and 28, with 21 animals per main group. Diabetes mellitus (DM) was induced by administration of streptozotocin (50 mg/kg via penile vein and 12-h fasting) and confirmed at day 21 (glycemic index > 240 mg/dL). In the animals of group P, 0.5 ml of the oil-water emulsion obtained from the plant seed was used. The samples were removed and hemisectioned, and one portion was used for the quantitative histological analysis of collagen using Masson's trichrome staining, while another portion was analyzed by FT-Raman spectroscopy. RESULTS: A higher percentage area of the volume of collagen fibers was observed for the experimental time Day 14 in group P compared with group D (p < 0.001). Regarding the ratio of areas of the amides I (1700‑1600 cm-1) and III (1245-1345 cm-1), the groups D and P show the opposite behavior. CONCLUSION: Cenostigma macrophyllum accelerated the repair process in skin of diabetic ratsfor14 days.

Study of advanced rheumatoid arthritis

INTRODUCTION: Rheumatic diseases are considered public health problems affecting millions of people worldwide resulting in high and rising health-care costs. In this work, Fourier Transform Infrared spectroscopy associated to Partial Least Square regression (PLS) analysis was used to diagnose rheumatoid arthritis (RA) from human serum. METHODS: The sera of 94 individuals were collected, which included 47 from rheumatic patients and 47 from healthy individuals. The results from PLS analysis were compared to standard clinical trials such as anti-citrullinated peptide antibodies, C- Reactive protein, and Rheumatoid factor. RESULTS: For clinical diagnosis, the anti-citrullinated peptide antibodies of second generation proved to be the most specific to diagnosis rheumatoid arthritis even after long periods of drug therapy. CONCLUSIONS: The qualitative PLS analysis has shown higher values of IgM of RA group, but the difference was very small. The RA patients were under medication, which interfered with the IgM concentration.

Effects of various hydrogen peroxide bleaching concentrations and number of applications on enamel

AIM: To evaluate the effects of three hydrogen peroxide (HP) concentrations on enamel mineral content (MC), following three peroxide applications and using three methods to measure the MC. METHODS : Forty samples were obtained from bovine incisors and randomly divided into 4 groups (n=10). The control group remained untreated, while the experimental groups were tested for three HP concentrations (10%, 35% and 50%). The HP gel was applied 3 times on the enamel surface for 30 min per application and the samples were analyzed after each application. The MC of the enamel was determined before and after bleaching using Fourier transform (FT-Raman) spectroscopy and micro energy-dispersive x-ray fluorescence spectrometry (µEDXRF). The calcium (Ca) lost from the bleached enamel was quantified with an atomic absorption spectrometer (AAS). The data were analyzed statistically by ANOVA, Tukey and Dunnett´s tests (p<0.05). RESULTS: The FT-Raman showed a decrease in MC for all bleaching treatments, without influence of the different HP concentrations or the number of applications. µEDXRF did not detect any changes in MC. CONCLUSIONS: Ca loss was observed by the AAS, with no difference among the three HP concentrations. The FT-Raman and AAS analyses detected MC reduction and Ca loss after HP bleaching.

Análise da composição bioquímica da pele por espectroscopia Raman / Skin biochemical composition analysis by Raman spectroscopy

O envelhecimento cutâneo caracteriza-se por alterações celulares e moleculares. Neste contexto, a espectroscopia Raman Confocal foi utilizada para medir estas alterações bioquímicas em função da profundidade da pele in vivo. Neste estudo houve a tentativa de correlacionar os modos vibracionais dos aminoácidos puros com os espectros da pele de voluntárias de diferentes faixas etárias. Foram coletados espectros in vivo de 32 voluntárias, sendo 11 para o Grupo A (20-23 anos), 11 para o Grupo B (39-42 anos) e 10 para o Grupo C (59-62 anos). Para cada grupo, os espectros Raman foram medidos na superfície (0 µm), 30 ± 3 µm e 60 ± 3 µm. Os resultados das comparações intergrupos mostraram que o grupo de maior idade teve prevalência da banda da tirosina, mas também apresentou um decréscimo da banda do ácido pirrolidônico, centrada em 875 cm-1. A banda da amida I, centrada em 1637 cm-1, atribuída à presença de colágeno, como também outras proteínas e lipídeos, apresentou uma menor intensidade no Grupo C, o que pode ser justificado pelo decréscimo na concentração de colágeno em função da idade.

Skin aging is characterized by cellular and molecular alterations. In this context, Confocal Raman spectroscopy was used in vivo to measure these biochemical changes as function of the skin depth. In this study we have tried to correlate spectra from pure amino acids to in vivo spectra from volunteers with different ages. This study was performed on 32 volunteers: 11 from Group A (20-23 years), 11 from Group B (39-42 years) and 10 from Group C (59-62 years). For each group, the Raman spectra were measured on the surface (0 µm), 30 ± 3 µm and 60 ± 3 µm below the surface. The results from intergroup comparisons showed that the oldest group had a prevalence of the tyrosine band, but it also presented a decrease in the band centered at 875 cm-1 of pyrrolidone acid. The amide I band centered at 1637 cm-1 that is attributed to collagen, as well as other proteins and lipid, showed a smaller amount of these biomolecules for Group C, which can be explained by the decrease in collagen concentration as a function of age.

Creatine Supplementation in Wistar Rats Submitted to Physical Tests of Swimming: Evaluation of the Physiological Effects through Raman Spectroscopy.

Creatine is a nitrogenous organic acid that naturally occurs in vertebrates. This compound is associated to energy supply to muscles, being that its biological synthesis occurs mainly in liver and is related to the aminoacids glycine, arginine and ornitine. Several studies have shown the benefits of creatine supplementation, such as higher muscle power, increased fat-free mass, and strength in healthy subjects. Besides, it can be applied as therapeutic agent for some diseases. In this work, the physical condition of Wistar rats that received creatine supplementation is compared with individuals without any supplementation. For this evaluation, swimming tests were developed in order to elucidate the physical condition of animals in different nutritional conditions. Indeed, swimming is considered one of the most complete exercises, since the practice of this physical activity involves great increase in the aerobic and anaerobic capability.

Influence of the photoinitiator system and light photoactivation units on the degree of conversion of dental composites

The aim of this study was to observe the influence of two light polymerization units (LED or halogen light) on the degree of conversion (DC) of three dental composites with lighter shades and a different photoinitiator system. The top (T) and bottom (B) surfaces of 60 discs of composite resin (Filtek™ Supreme, Filtek™ Z250, Tetric™ Ceram Bleach) cured either by LED or by halogen lamp (HL) were studied using an FT-Raman spectrometer. The degree of conversion (DC) was evaluated by following the changes in the intensity of the methacrylate C=C stretching mode at 1640 cm-1. The calculated DC ranged from 54.2 percent (B) to 73.4 percent (T) and from 60.2 percent (B) to 76.6 percent (T) for the LED and HL, respectively. LED and halogen devices were able to produce an adequate DC for all the resins tested.

Estudo da ação do ultrassom terapêutico em modelo experimental de tendinite em ratos Wistar / Study of the therapeutic efficacy of ultrasound in experimental model of tendinitis in Wistar rats

Introdução: O ultrassom terapêutico acelera o reparo do tendão. Objetivos: Estudar a ação do ultrassom em tendinite experimental por análises histomorfométrica e histopatológica. Método: Vinte e cinco ratos Wistar, 220-250 g, foram divididos em três grupos experimentais (7 e 14 dias): A7 e A14, tendinite induzida sem tratamento; B7 e B14, tendinite induzida tratada com ultrassom pulsado de 1 MHz a 10% 0, 5 W/cm², e C, o grupo sem indução de tendinite. Resultados: A histomorfometria mostrou-se altamente significativa (p <0,001) do ultrassom em contra-relógio de 14 dias, comparado com o grupo saudável (C), e muito significativamente em comparação com o mesmo grupo tendinite (A7 e A14) (p <0,01). Conclusão: A histologia e a histomorfometria demonstraram qualitativa e quantitativamente, um aumento do número de fibroblastos, em sete dias, e das fibras de colágeno, em 14 dias, para o grupo tratado com ultrassom em relação ao tendinite.

Introduction: The therapeutic ultrasound accelerates repair of tendon. Objectives: Analyze the action of ultrasound in experimental tendonitis in histomorphometric and histopathological analysis. Methods: Twenty-five Wistar rats, 220 ? 250 g, were divided into three experimental groups (7 and 14 days): A7 and A14 induced tendinitis and without treatment; B7 and B14, induced tendinitis and treated with pulsed ultrasound 1 MHz to 10% 0.5 W/cm², and C, the group without induction of tendinitis. Results: Histomorphometry was highly significant (p <0.001) of ultrasound against the clock for 14 days, compared with the healthy group (C), and very significantly in relation to the same group tendinitis (A7 and A14) (p <0, 01). Conclusion: The histology and histomorphometry demonstrated qualitatively and quantitatively, an increased number of fibroblasts in seven days, and the collagen fibers, in 14 days, for the group of ultrasound compared with the group of tendinitis.

Differential diagnosis in primary and metastatic cutaneous melanoma by FT-Raman spectroscopy / Diagnóstico diferencial no melanoma primário e metastático por espectroscopia FT-Raman

PURPOSE: To qualify the FT-Raman spectral data of primary and metastatic cutaneous melanoma in order to obtain a differential diagnosis. METHODS: Ten normal human skin samples without any clinical or histopathological alterations, ten cutaneous melanoma fragments, and nine lymph node metastasis samples were used; 105, 140 and 126 spectra were obtained respectively. Each sample was divided into 2 or 3 fragments of approximately 2 mm³ and positioned in the Raman spectrometer sample holder in order to obtain the spectra; a monochrome laser light Nd:YAG at 1064 nm was used to excite the inelastic effect. RESULTS: To differentiate the three histopathological groups according to their characteristics extracted from the spectra, data discriminative analysis was undertaken. Phenylalanine, DNA, and Amide-I spectral variables stood out in the differentiation of the three groups. The percentages of correctly classified groups based on Phenylalanine, DNA, and Amide-I spectral features was 93.1 percent. CONCLUSION: FT-Raman spectroscopy is capable of differentiating melanoma from its metastasis, as well as from normal skin.

OBJETIVO: Qualificar os dados espectrais FT-Raman do melanoma cutâneo primário e metastático e assim realizar o diagnóstico diferencial. MÉTODOS: Foram utilizadas amostras de 10 fragmentos de pele sem alterações clínicas ou histopatológicas, 10 de melanomas cutâneos e 9 de metástases linfonodais; 105, 140 and 126 espectros foram obtidos respectivamente. Cada amostra foi dividida em 2 ou 3 frações de 2 mm³ e posicionada no porta amostras do espectrômetro Raman para obtenção dos espectros, por meio da excitação do espalhamento inelástico pelo laser de Nd:YAG em 1064 nm incididos na amostra. RESULTADOS: Para diferenciar os três grupos formados de acordo com as características fornecidas pelos espectros, realizamos a análise discriminante dos dados. As variáveis espectrais Fenilalanina, DNA e Amida-I se destacaram na capacidade de diferenciação dos três grupos histológicos. A porcentagem de classificação correta utilizando estes critérios foi de 93,1 por cento; o que mostra a eficiência da análise realizada. CONCLUSÃO: A espectroscopia FT-Raman é capaz de diferenciar o melanoma de sua metástase, assim como da pele normal.

FT-Raman spectroscopy for the differentiation between cutaneous melanoma and pigmented nevus / Espectroscopia FT-Raman na diferenciação entre melanoma cutâneo e nevo pigmentado

Cutaneous melanoma is the most aggressive type of skin cancer and Ft-Raman spectroscopy has been studied as a potential method that could be a real alternative for early diagnosis of neoplasms. PURPOSE: To qualify the spectral FT-Raman data, in order to differentiate cutaneous melanoma and pigmented nevus. METHODS: For this study, 10 samples of cutaneous melanoma, 9 samples of pigmented nevi, and 10 samples of normal skin were obtained by incisional biopsies performed during plastic surgeries ex vivo, immediately after removing the surgical sample. RESULTS: The FT-Raman spectra of each group presented a high correlation between the elements of the same group, thus favoring the elaboration of spectral averages. When analyzing the spectral standard of each group, the normal skin standard did not show a significant variation between the spectra; the standard of the pigmented nevi group showed significant variation, and the cutaneous melanoma group also showed variation. Through univariate analysis, specific bands were detected for each vibrational mode identified. The discriminatory analysis of the data showed a 75.3 percent efficiency of the differentiation between the three groups studied. CONCLUSION: The vibrational modes Polysaccharides, Tyrosine and Amide-I differentiated the melanoma from the pigmented nevus.

O melanoma cutâneo é o câncer de pele mais agressivo, e a espectroscopia FT-Raman tem sido estudada como um método em potencial que pode ser uma verdadeira alternativa no diagnóstico precoce de neoplasias. OBJETIVO: Qualificar os dados espectrais FT-Raman de modo a diferenciar melanoma cutâneo de nevo pigmentado. MÉTODOS: Foram utilizadas 10 amostras de melanoma cutâneo, obtidas por meio de biopsias incisionais realizadas "ex-vivo"; nove amostras de nevo pigmentado e 10 amostras de pele normal foram coletadas durante cirurgias plásticas. RESULTADOS: Os espectros FT-Raman de cada grupo diagnóstico apresentaram alta correlação entre os elementos do mesmo grupo, o que favoreceu a realização das médias espectrais. Analisando o padrão espectral de cada grupo, o de pele normal não mostrou grande variação entre os espectros; o de nevo pigmentado apresentou variação notável e, o grupo melanoma primário também indicou variação. Por meio de análise univariada foram identificadas bandas específicas para cada modo vibracional identificado. A análise discriminante aos dados mostrou 75,3 por cento de eficiência na diferenciação entre os três grupos estudados. CONCLUSÃO: Os modos vibracionais Polissacarídeos (Banda I), Tirosina (Banda 6) e Amida I (Banda 10) diferenciaram o melanoma do nevo pigmentado.