Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization

Abstract The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. Objective To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. Methodology Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. Results The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). Conclusion Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

Role of Candida albicans on enamel demineralization and on acidogenic potential of Streptococcus mutans in vitro biofilms

Abstract There is growing evidence that C. albicans is associated with dental caries, but its role on caries development needs to be better clarified. Objective: To evaluate at the hard tissue level the effect of C. albicans on the cariogenic potential of S. mutans biofilms focusing on the mineral profile of induced carious lesions. This study also aimed to evaluate the effect of C. albicans on the acidogenic potential of S. mutans biofilms. Methodology: Dual-species (CA+SM) and single-species biofilms (CA or SM) were grown on the surface of enamel slabs in the presence of glucose/sucrose supplemented culture medium for 24, 48 and 72 hours. Demineralization was evaluated through percentage of surface microhardness change (%SMC) and transversal microradiography analysis (ILM and LD) and pH of the spent medium was recorded daily. Data were analyzed by two-way ANOVA followed by Bonferroni correction. Results: %SMC was statistically different among the biofilms at each time point being the highest for SM biofilms and the lowest for CA biofilms which also differed from CA+SM biofilms [SM (24 h: 47.0±7.3; 48 h: 66.3±8.3; 72 h: 75.4±3.9); CA (24 h: 7.3±3.3; 48 h: 7.1±6.4; 72 h: 6.6±3.6); CA+SM (24 h: 35.9±7.39.1; 48 h: 47.2±9.5; 72 h: 47.6±9.5)]. pH of spent medium was statistically lower for SM biofilms compared to the other biofilms at each time point and remained constant over time while pH values increased from 24 to 72 h for both CA and CA+SM biofilms [SM (24 h: 4.4±0.1; 48 h: 4.4±0.1; 72 h: 4.5±0.1); CA (24 h: 6.9±0.3; 48 h: 7.2±0.2; 72 h: 7.5±0.2); CA+MS (24 h: 4.7±0.2; 48 h: 5.1±0.1; 72 h: 6.1±0.6)]. IML and LD for SM biofilms increased over time while no difference was observed from 24 to 72 h for the other biofilms. Conclusions: The present data suggest that C. albicans has low enamel demineralization potential and the presence of C. albicans can reduce both the cariogenic and acidogenic potentials of S. mutans biofilms.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization

Abstract Objectives: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. Methodology: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). Results: M. urundeuva All. at 100, 10 and 0.1 μg/mL and Q. grandiflora Mart. at 100 and 0.1 μg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 μg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 μg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. Conclusions: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Comparison between static and semi-dynamic models for microcosm biofilm formation on dentin

Abstract Microcosm biofilm has been applied to induce carious lesions in dentin. However, no study has been done to compare the impact of the type of model for providing nutrients to microcosm biofilm formation on dentin. Objective This study compared the performance of two kinds of models (static and semi-dynamic) on the biofilm formation and the development of dentin carious lesions. Material and Methods In both models, biofilm was produced using inoculum from pooled human saliva mixed with McBain saliva for the first 8 h (5% CO2 and 37°C). Afterwards, for the static model, the samples were placed in 24-wells microplate containing McBain saliva with 0.2% sucrose, which was replaced at 24 h. In the semi-dynamic model, the samples were submitted to artificial mouth system with continuous flow of McBain saliva with 0.2% sucrose (0.15 ml/min, 37°C) for 10 h a day (for the other 14 h, no flow was applied, similarly to the static model). After 5 days, biofilm viability was measured by fluorescence and dentin demineralization by transverse microradiography. Results Biofilm viability was significantly lower for the static compared with semi-dynamic model, while dentin demineralization was significantly higher for the first one (p<0.05). The static model was able to produce a higher number of typical subsurface lesions compared with the semi-dynamic model (p<0.05). Conclusions The type of model (static and semi-dynamic) applied in the microcosm biofilm may have influence on it's viability and the severity/profile of dentin carious lesions.

Effect of Myracrodruon urundeuva and Qualea grandiflora extracts on viability and activity of microcosm biofilm and prevention of enamel demineralization in vitro / Efeito de extratos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade e atividade de biofilme microcosmo e na prevenção da desmineralização do esmalte in vitro

The objective of this study was to evaluate the antimicrobial and anti-caries effects of two plant extracts. The first chapter dealt with a review of the literature whose objective was to discuss the antimicrobial potential of Brazilian natural agents on the biofilm related to dental caries and gingivitis/periodontal disease. The research of the articles was carried out using PubMed. We found a total of 23 papers. Most of the studies were performed using planktonic microorganisms or under clinical trials. Nineteen articles were focused on cariogenic bacteria. From these nineteen articles, eleven were also about periodontopathogenic bacteria. Four studies addressed only periodontopathogenic bacteria. The most tested Brazilian natural agents were green propolis, essential oils of Lippia sidoides and Copaifera sp. Most of the tested agents showed similar results when compared to positive control (essential oils and extracts) or better effect than negative control (green propolis). More studies involving protocols closer to the clinical condition and the use of response variables that allows understanding the mechanism of action of natural agents are necessary before the incorporation of these natural agents into dental products. The second chapter aimed to test the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability of the microcosm biofilm and on the prevention of enamel demineralization. The microcosm biofilm was produced on bovine enamel, using human saliva pool mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva at 100, 10 and 0.1 µg/ml and Q. grandiflora at 100 and 0.1 µg/ml reduced cell viability similarly to the positive control and significantly more than negative control. M. urundeuva at 1000, 100 and 0.1 µg/ml were able to reduce the counting formation unit-CFU counting of lactobacilli sp. and Streptococcus mutans, while Q. grandiflora at 1000 and 1.0 µg/ml significantly reduced the S. mutans CFU counting. On the other hand, the natural extracts did not reduce the production of extracellular polyssacharides, lactic acid and the development of enamel caries lesions. The third chapter aimed to evaluate the effect of hydroalcoholic extracts of M. urundeuva and Q. grandiflora (alone or combined) on the viability of S. mutans biofilm and the prevention of enamel demineralization. S. mutans strain (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration and minimum biofilm eradication concentration were determined to choose the concentrations to be tested under the biofilm model. S. mutans biofilm (5x105 CFU/ml) was produced on bovine enamel using McBain saliva with 0.2% sucrose for 3 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva (isolated or combined) at concentrations equal or higher than 0.625 mg/ml was able to reduce the bacteria viability, whereas Q. grandiflora extract alone showed antimicrobial effect at 5 mg/ml only (p<0.05). On the other hand, none of the extracts was able to reduce the development of enamel caries lesions. Despite the tested natural extracts have antimicrobial effect; they are unable to prevent caries in enamel.(AU)

O objetivo foi avaliar os efeitos antimicrobiano e anti-cárie de dois extratos de plantas. O primeiro capítulo se referiu a uma revisão da literatura cujo objetivo foi discutir o potencial antimicrobiano dos agentes naturais brasileiros sobre o biofilme relacionado à cárie dentária e à gengivite/doença periodontal. A pesquisa dos artigos foi realizada usando o PubMed. Foram encontrados 23 trabalhos. A maioria dos estudos foi realizada utilizando microorganismos na fase planctônica ou ensaios clínicos. Dezenove artigos foram focados em bactérias cariogênicas. Dos dezenove artigos, onze também eram sobre bactérias periodontopatogênicas. Quatro estudos abordaram apenas bactérias periodontopatogênicas. Os agentes naturais brasileiros mais testados foram própolis verde, óleos essenciais de Lippia sidoides e Copaifera sp. Os agentes testados apresentaram resultados similares quando comparados ao controle positivo (óleos essenciais e extratos) ou melhor efeito que o controle negativo (própolis verde). Mais estudos próximos da condição clínica e o uso de variáveis de resposta que permitam entender o mecanismo de ação são necessários, para permitir a incorporação desses agentes naturais em produtos odontológicos. O segundo capítulo teve como objetivo testar o efeito dos extratos hidroalcoólicos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade do biofilme microcosmo e na prevenção da desmineralização do esmalte. O biofilme microcosmo foi produzido em esmalte bovino, utilizando pool de saliva humana misturada à saliva de McBain (0,2% de sacarose) durante 14 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva a 100, 10 e 0,1 µg/ml e Q. grandiflora a 100 e 0,1 µg/ml reduziram a viabilidade dos microrganismos de forma semelhante ao controle positivo e significativamente maior do que o controle negativo. M. urundeuva a 1000, 100 e 0,1 µg/ml foi capaz de reduzir a contagem de Unidade formadora de colônia-UFC para Lactobacilos totais e Streptococcus mutans, enquanto a Q. grandiflora a 1000 e 1,0 µg/ml reduziu significativamente a contagem de UFC para S. mutans. Os extratos naturais não conseguiram reduzir a produção de polissacarídeos extracelulares-PEC, ácido lático e o desenvolvimento da lesão cariosa em esmalte. O terceiro capítulo teve como objetivo avaliar o efeito dos extratos hidroalcoólicos de M. urundeuva. e Q. grandiflora (sozinhos ou combinados) sobre a viabilidade do biofilme de S. mutans e na prevenção da desmineralização do esmalte. Cepa de S. mutans (ATCC 21175) foi reativada em caldo BHI. Concentração inibitória mínima, concentração bactericida mínima, concentração inibitória mínima de biofilme e concentração de erradicação mínima de biofilme foram determinadas para escolher as concentrações a serem testadas sob o modelo de biofilme. O biofilme de S. mutans (5x105 CFU/ml) foi produzido em esmalte bovino, utilizando saliva de McBain com 0,2% de sacarose durante 3 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva (isolada ou combinada) nas concentrações iguais ou superiores a 0,625 mg/ml foi capaz de reduzir a viabilidade das bactérias, enquanto que o extrato da Q. grandflora apresentou efeito antimicrobiano somente a 5 mg/ml (p<0,05). Nenhum dos extratos reduziu o desenvolvimento da lesão da cárie. Apesar dos extratos naturais terem efeito antimicrobiano, são incapazes de prevenir o desenvolvimento da lesão cariosa em esmalte.(AU)

Analysis of the antimicrobial and anti-caries effects of TiF4 varnish under microcosm biofilm formed on enamel

Abstract Titanium tetrafluoride (TiF4) is known for interacting with enamel reducing demineralization. However, no information is available about its potential antimicrobial effect. Objectives This study evaluated the antimicrobial and anti-caries potential of TiF4 varnish compared to NaF varnish, chlorhexidine gel (positive control), placebo varnish and untreated (negative controls) using a dental microcosm biofilm model. Material and Methods A microcosm biofilm was produced on bovine enamel previously treated with the varnishes, using inoculum from human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. All experiments were performed in biological triplicate (n=4/group in each experiment). Factors evaluated were: bacterial viability (% dead and live bacteria); CFU counting (log10 CFU/mL); and enamel demineralization (transverse microradiography - TMR). Data were analysed using ANOVA/Tukey's test or Kruskal-Wallis/Dunn's test (p<0.05). Results Only chlorhexidine significantly increased the number of dead bacteria (68.8±13.1% dead bacteria) compared to untreated control (48.9±16.1% dead bacteria). No treatment reduced the CFU counting (total microorganism and total streptococci) compared to the negative controls. Only TiF4 was able to reduce enamel demineralization (ΔZ 1110.7±803.2 vol% μm) compared to both negative controls (untreated: ΔZ 4455.3±1176.4 vol% μm). Conclusions TiF4 varnish has no relevant antimicrobial effect. Nevertheless, TiF4 varnish was effective in reducing enamel demineralization under this model.

Comparação de diferentes pré-tratamentos de superfície na remineralização de esmalte com flúor em tratamento de lesões com mancha branca e sua resistência após desafio ácido / Comparison of different pretreatments on remineralization of enamel white spot lesions with fluoride and its resistance after new acidic challenge

A aplicação de flúor tópico é a principal estratégia de natureza química para a remineralização de lesões incipientes (LI) clinicamente visíveis como manchas brancas (MB) por cárie. Abordagens para aumentar a retenção de F no substrato pode favorecer sua ação e para isso, tratamentos prévios da superfície do esmalte podem ser usados. O objetivo deste trabalho foi comparar a capacidade de remineralização do flúor sem e com pré-tratamento do esmalte com ácido fosfórico e nitrato de alumínio e a sua resistência após novo desafio. Sessenta espécimes de esmalte bovino foram preparados (6mm x 4mm) e selecionados por meio de análise de microdureza de superfície (MS). LI foram produzidas através de ciclagem Desmineralização-Remineralização (DES-RE) e os espécimes divididos aleatoriamente em cinco grupos, de acordo com o tratamento (n=12): V- controle (verniz de fluoreto de sódio 5% por 4 horas), F (sem pré-tratamento); P-F(condicionamento ácido com ácido fosfórico por 30s); Al-F (nitrato de alumínio a 0,05M por 1min); P-Al-F (condicionamento com ácido fosfórico a 37% + nitrato de alumínio a 0,05M). Os tratamentos foram repetidos semanalmente durante quatro semanas. Após o tratamento, os espécimes foram submetidos à nova ciclagem ácida. Após cada etapa, nova MS foi realizada e ao final das análises, um corte transversal dos espécimes foi realizado. Uma das metades foi submetida à análise da microdureza longitudinal (ML) e a outra preparada para realização da microrradiografia transversal (TMR) para a análise de conteúdo mineral perdido. O percentual de perda de dureza de superfície (%PDS) foi analisado por ANOVA a 2 critérios de medidas repetidas e Tukey e o percentual de perda de dureza longitudinal (%PDL) por ANOVA a 3 critérios de medidas repetidas e Tukey (p<0,05). Os dados de TMR no parâmetro LD (profundidade da lesão) foram analisados por ANOVA a dois critérios (p<0,05). Os resultados mostraram que a ciclagem DES-RE resultou em significante %PDS e %PDL em todos os grupos. Grupo F revelou menor perda de MS após tratamento e, F, Al-F e P-Al-F, mostraram menor perda de dureza final, após o novo desafio ácido. Na análise de %PDL, o grupo V apresentou menor perda de dureza final quando comparado com os demais grupos, nas diferentes profundidades. A análise do conteúdo mineral não revelou nenhuma diferença entre os tratamentos e fases. Nenhum dos pré tratamentos propostos foram capazes de otimizar a atuação do gel APF na remineralização de MB.(AU)

The topical fluoride is the main chemical strategy to remineralize incipient caries lesions (ICL) visible as carious white spot lesions (WSL). Approaches to increase the fluoride retention may favor its action and therefore, enamel pretreatments can be used for this purpose. The aim of this study was to compare the potential of fluoride remineralization with and without previous enamel pretreatment with aluminum nitrate and phosphoric acid as well as their resistance after a new acid challenge. Sixty bovine enamel specimens were prepared (6mm x 4mm) and selected by the surface hardness (SH) analysis. The ICL were produced using DE-RE cycling and the specimens were randomized in five groups, according to the treatment (n=12): Vcontrol (5% sodium fluoride varnish during 4h), and four groups previously preatreated with topical application of acidulated phosphate fluoride (APF) during 4min: F (without pretreatment), P-F (phosphoric acid etching during 30s), Al-F (0.05M aluminum nitrate during 1min); P-Al-F (phosphoric acid etching + aluminum nitrate). The treatments were performed weekly during four weeks. After the treatment, the specimens were submitted to the new acid challenge. After each step, a new SH analysis was performed followed by the transversal cut of the specimens. A half was submitted to the longitudinal cross-sectional hardness analysis (LH) and the other half was prepared to transverse microradiography assessments (TMR) to determine the loss of mineral content. The percentage of surface mineral loss (%SH) was analyzed using two-way repeated-measures ANOVA and the percentage of crosssectional mineral loss (%LH) by the three-way repeated-measures ANOVA and Tukey (p<0.05). Data of TMR analysis by LD (lesion depth) parameter was analyzed with two-way ANOVA (p<0.05). The results showed significant %SH and %LH after DE-RE cycling for all groups. F group showed the lowest %SH after treatment, with no significant difference to Al-F and P-Al-F after the new acid challenge. V group showed the lowest %LH compared to the other groups, in the different depths, suggesting more resistance. Mineral content assessment did not reveal any difference among treatments and phases. None pretreatments were able to increase the potential of fluoride remineralization.(AU)

Assessment of Early Dental Caries by Using Optical Coherence Tomography / 치위생과학회지

The purpose of this study was to assess the correlation between integrated mineral loss (volume % mineral×µm, ΔZ(TMR)) determined using transverse microradiography (TMR) and integrated reflectivity (dB×µm, ΔR(OCT)) determined using optical coherence tomography (OCT) for detecting early dental caries with lesion depth more than 200 µm. Sixty tooth specimens were made from sound bovine teeth. They were immersed in a demineralized solution for 20, 30, and 40 days. The ΔR(OCT) was obtained from the cross-sectional OCT image. The ΔZ(TMR) was obtained from the TMR image. The correlation between ΔR(OCT) and ΔZ(TMR) was examined using Pearson correlation. The Bland-Altman plot was constructed using the ΔR(OCT) and ΔZ(TMR) values. A significant correlation between ΔR(OCT) and ΔZ(TMR) was confirmed (r=0.491, p=0.003). Moreover, most of the difference between ΔR(OCT) and ΔZ(TMR) was included in the error section of the Bland-Altman plot. Therefore, OCT could be used as a substitute for TMR when analyzing mineral loss in early dental caries.

Effect of 10% fluoride on the remineralization of dentin in situ

ABSTRACT Objective The purpose of this randomized, cross-over, in situ study was to determine the remineralization of demineralized dentin specimens after the application of a 10% fluoride (F-) or a 1% chlorhexidine–1% thymol (CHX–thymol) varnish. Material and Methods Twelve individuals without current caries activity wore removable appliances in the lower jaw for a period of four weeks. Each appliance contained four human demineralized dentin specimens fixed on the buccal aspects. The dentin specimens were obtained from the cervical regions of extracted human third molars. After demineralization, half the surface of each specimen was covered with a nail varnish to serve as the reference surface. The dentin specimens were randomly assigned to one of the three groups: F-, CHX–thymol, and control (no treatment). Before the first treatment period and between the others, there were washout periods of one week. After each treatment phase, the changes in mineral content (vol% µm) and the lesion depths (µm) of the dentin slabs were determined by transverse microradiography (TMR). Data analysis was accomplished by the Kruskal-Wallis test and the Mann-Whitney U test (p<0.05). Results The medians (25th/75th percentile) of integrated mineral loss were 312.70 (203.0-628.7) for chlorhexidine varnish, 309.5 (109.8-665.8) for fluoride varnish, and -346.9 (-128.7 - -596.0) for the control group. The medians (25th/75th percentile) of lesion depth were 13.6 (5.7-34.5) for chlorhexidine varnish, 16.5 (5.6-38.1) for fluoride varnish, and -14.2 (-4.5- -32.9) for the control group. Use of the 10% F- or 1% CHX–1% thymol varnishes resulted in significantly decreased mineral loss and lesion depth in dentin when compared with the control group. There were no statistically significant differences among the test groups. Conclusions Within the limitations of this study, the results suggest that the effect of the treatment of demineralized dentin with 10% F- or 1% CHX–1% thymol is better than without any treatment.

Cariostatic effect of various cheeses on the degree of mineralisation of enamel in situ

To investigate the effect of 9 cheeses with different compositions on the extent of demineralisation/ remineralisation of enamel slabs in situ. Sucrose and sorbitol solutions were used as controls. Ten subjects were required to wear removable appliances with human enamel slabs cut from white spot lesions created in vitro. The subjects were required to wear their appliances, for two days to allow plaque to grow. Over the following 5 days, they were asked to immerse their appliances 4 times per day for 10 minutes each time in a suspension of the test cheese or control solution. De- or remineralisation was then measured using a combination of microradiography and an image analysis system. The results for lesion depth [ micro m] tested by paired t-test revealed significant net remineralisation [p<0.05] with two types of cheeses. As for mineral loss or gain [vol.% micro m], significant net remineralisation was seen with 6 types of cheeses. Some cheeses can cause remineralisation of early lesions

Remineralization of carious dentin. II: In vivo microradiographic and chemical studies in human permanent teeth capped with calcium hydroxide

O principal objetivo deste estudo in vivo foi verificar uma possível remineralização da dentina cariada humana. Oitenta e seis amostras de dentina cariada foram estudadas, removidas de 36 dentes permanentes de 24 pacientes e divididas em não-tratadas (controles) e capeadas com hidróxido de cálcio quimicamente puro (experimentais), estudadas de acordo com períodos de observação entre 10 e 120 dias, classificadas levando-se em conta profundidade e grau de amolecimento dentinário e avaliadas em relação a peso, concentração de fósforo, microradiografias qualitativas e quantitativas e valores absolutos de conteúdo mineral total. As amostras experimentais foram examinadas ao microscópio óptico e um aumento qualitativo em radiopacidade foi observado. Quantitativamente, foi mostrado que, no caso das amostras analisadas para concentração de fósforo, a média das diferenças em aumento de porcentagens após tratamento foi da ordem de 9.56%, enquanto que para as amostras avaliadas microradiograficamente o aumento do conteúdo mineral total foi de 22.29%. Em ambos os casos, as diferenças registradas foram estatisticamente significantes.

Remineralization of carious dentin. I: In vitro microradiographic study in human teeth capped with calcium hydroxide

O objetivo principal deste estudo in vitro foi o de verificar uma possível remineralização da dentina cariada humana. Trinta e nove dentes permanentes e deciduos recém-extraídos foram utilizados. Cada dente foi dividido em duas metades. Uma metade foi usada como controle e a outra como experimental. Nesta última, uma cavidade foi preparada e a camada remanescente de dentina desmineralizada capeada com hidróxido de cálcio quimicamente puro. As amostras experimentais foram armazenadas em estufa a 37ºC. Os períodos de observação foram de 2, 4, 6, 8, 10 e 12 semanas. Todas as metades foram desgastadas até conseguir-se cortes plano-paralelos com espessuras variando entre 75 e 117 µm. Microradiografias qualitativas evidenciaram um aumento na radiopacidade das amostras tratadas com hidróxido de cálcio. A microradiografia quantitativa evidenciou um aumento estatisticamente significante no conteúdo mineral total nas amostras experimentais comparadas com as controles. Os resultados indicam uma remineralização in vitro da dentina cariada através do hidróxido de cálcio.

Volume Fraction of the Human Pathologic Ossicles Destructed by Cholesteatoma Using Micro-CT / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Conventional studies for the middle ear ossicles destructed by cholesteatoma give relatively limited information on shape, mechanical strength and 3-dimensional relationship. Recently, Micro-CT scanning technology has been applied to analysis of skeletal structure. This study aims to reconstruct the 3-dimensional structure of normal ossicles and destructed ossicles by different cholesteatomas and to compare volume fraction of destructed ossicles with that of normal ossicles. MATERIALS AND METHOD: 13 pathological ossicles obtained during cholesteatoma surgery were used in this study (2 malleuses and 2 incuses from attic cholesteatoma;1 malleus and 2 incuses from sinus cholesteatoma;2 malleuses and 2 incuses from tensa retraction cholesteatoma; 2 incuses from congenital cholesteatoma). As a control, normal ossicles harvested from cadaveric temporal bone specimens were used. The structure of obtained ossicles was analyzed through Micro-CT (SkySan-1072, Belgium) by filming the 2-dimensional cross section image running perpendicular to the long axis of the ossicles. The 3-dimensional images were thereby reconstructed for each, and were analyzed using CT-Analyzer(TM) and ANTTM. Based on the results, the strength of each ossicle were measured. RESULTS: The volume fraction, indicating relative strengths of ossicles, was lower in the portion of destructed surface. The pattern of destructed ossicles by cholesteatoma was different according to pathological status of cholesteatoma. CONCLUSION: Our preliminary data of ossicular structure analyzed by Micro-CT scan will be helpful for elucidation of pathological ossicles in middle ear cholesteatoma and will make a great contribution to research of middle ear cholesteatoma.

In vivo quantitative analysis of remineralization effect of remineralization solution "R" of incipient enamel dental caries / 대한치과보존학회지

Dental caries is a chronic disease that causes the destruction of tooth structure by the interaction of plaque bacteria, food debris, and saliva. There has been attempts to induce remineralization by supersaturating the intra-oral environment around the surface enamel, where there is incipient caries. In this study, supersaturated remineralized solution "R" was applied to specimens with incipient enamel caries, and the quantitative ananlysis of remineralization was evaluated using microradiography. Thirty subjects volunteered to participate in this study. Removable appliances were constructed for the subjects, and the enamel specimen with incipient caries were embedded in the appliances. The subjects wore the intra-oral appliance for 15 days except while eating and sleeping. The removable appliance were soaked in supersaturated solution "R", saline, or Senstime(R) to expose the specimen to those solutions three times a day, 5 minutes each time. After 15 days, microradiography was retaken to compare and evaluate remineralization. The results were as the following: 1. The ratio of remineralized area to demineralized area was significantly higher in the supersaturated solution "R" and Senstime(R) than in the saline. (p<0.05) 2. Remineralization in the supersaturated buffer solution "R" occurred in the significantly deeper parts of the tooth, compared to the Senstime(R) group containing high concentration of fluoride.(p<0.05) As in the above results, the remineralization effect of remineralized buffer solution "R" on incipient enamel caries has been proven. For clinical utilization, further studies on soft tissue reaction and the effect on dentin and cementum are necessary. In conclusion compared to commercially available fluoride solution, remineralization solution "R" showed better remineralization effect on early enamel caries lesion, so it is considered as effecient solution for clinical application.

The Experimental Study Of Effects Of Irradiation On Osseointegration / 대한구강악안면방사선학회지

The purpose of this study was to investigate the effects of the Co-60 gamma irradiation on the osseointegration. 2.0 mm titanium alloy screw implants were placed in the tibial metaphysis of the rabbits, bilaterally. The mean length of the implants was 6.0 mm. The right tibia was irradiated with a single dose of 15 Gy from 60Co teletherapic machine at 5th postoperative day. The experimental group was irradiated tibia. The control group was non-irradiated tibia. To observe the phase of bone formation, the bone labeling by intramuscular injection of 20mg/Kg of Tetracycline, Calcein, Alizarin red S, was performed. The rabbits were sacrificed on the 1st, 2nd, 4th, 6th, 8th week and the tibia including implants were taken, and then the specimens were examined by the microradiography, light microscopy, and fluorescent microscopy. The obtained results were as follows; 1. There were connective tissue between bone and titanium at 1st week, in both group. Especially, the many empty lacunae without nucleus and obscure cytoplasm in experimental group, were observed. 2. The osteons were observed at 4th week in control group, and at 6th week in experimental group. The bone formation in experimental group was retarded as compared to the control group. 3. In fluorescent microscopy, bone labelling band was observed as linear, arc or concentric shape. Occasionary interrupted labelling band was observed, which is demonstrated bone remodeling. 4. In microradiographic examination, the radiolucent image was found between bone and implant with widening of bone marrow spaces as compared to the control group.

Effect of a specially formulated remineralizing solution on the remineralizing capacity of saliva.

The present study was carried out to assess whether the remineralization of artificial caries like lesion by artificial saliva can be improved by daily 5 minute treatment in a specially formulated remineralizing solution containing Ca, P, Zn, Sr, F. Extracted human premolars were dipped for 24 hours in artificial saliva and subsequently daily for 5 minutes in the remineralizing solution for 3 weeks. Lesions were assessed using transverse microradiography, electron probe microanalysis (EPMA) and scanning electron microscope (SEM). The results showed that remineralization by specially formulated solutions repair the enamel lesion and enhances the salivary remineralizing potential.

Aplicaçäo de esforços de torçäo no osso "in vivo" / Torsional forces applied to the living bone

Um fixador externo especial foi confeccionado de modo que sua extremidade distal pudesse ser acionada para rodar ao longo de um eixo central. Foi aplicado na face medial da tíbia direita de cordeiros de seis meses de idade por meio de quatro pinos de Schanz e regulado diariamente de maneira de aplicar um torque de 32 kgf.cm, até que produzisse torçao externa de 30§ na extremidade distal da tíbia, sendo, depois disso, mantido por três meses adicionais. Por meio de ponte de extensometria, foi possível monitorar o torque durante a aplicaçao da torçao e após este período. Após o sacrifício dos animais, foi encontrado que o osso deformou-se permanentemente a favor da aplicaçao da torçao, quando comparado com a tíbia normal. Por meio de microrradiografias, microscopia de luz polarizada e luz ultravioleta, foi constatado que houve modificaçao na microestrutura do osso, com áreas de deposiçao óssea assimétrica que mudaram o formato circular da secçao transversa da diáfise da tíbia para uma forma elipsóide.

Sugestäo de metodologia para avaliaçäo da interface implante/tecido ósseo / Suggestion of a methodology to evaluate the implant/bone tissue interface

Com o objetivo de desenvolver uma metodologia simples e de baixo custo para o estudo da interface implante/tecido ósseo, pinos de aço inoxidável revestidos com Al2O3, TiO2, Nb2O5 e sem revestimento foram implantados em fêmures de cäes. Após o óbito dos animais, as amostras foram radiografadas por técnica convencional (que se mostrou imprópria), técnica de alta resoluçäo (que permitiu boa avaliaçäo preliminar), microrradiografadas e analisadas por espectromia de fluorescência de raios X, o que possibilitou a avaliaçäo da liberaçäo de material pelos implantes. As microrradiografias tiveram sua densidade óptica medida, o que permitiu a determinaçäo precisa da espessura do tecido formado ao redor dos implantes.