Comparison of in vivo and in vitro models to evaluate pulp temperature rise during exposure to a Polywave® LED light curing unit

Abstract Objectives: To measure and compare in vivo and in vitro pulp temperature (PT) increase (ΔTEMP) over baseline, physiologic temperature using the same intact upper premolars exposed to the same Polywave® LED curing light. Methodology: After local Ethics Committee approval (#255,945), local anesthesia, rubber dam isolation, small occlusal preparations/minute pulp exposure (n=15) were performed in teeth requiring extraction for orthodontic reasons. A sterile probe of a temperature measurement system (Temperature Data Acquisition, Physitemp) was placed within the pulp chamber and the buccal surface was sequentially exposed to a LED LCU (Bluephase 20i, Ivoclar Vivadent) using the following exposure modes: 10-s low or high, 5-s Turbo, and 60-s high. Afterwards, the teeth were extracted and K-type thermocouples were placed within the pulp chamber through the original access. The teeth were attached to an assembly simulating the in vivo environment, being similarly exposed while real-time temperature (°C) was recorded. ΔTEMP values and time for temperature to reach maximum (ΔTIME) were subjected to two-way ANOVA and Bonferroni's post-hoc tests (pre-set alpha 0.05). Results: Higher ΔTEMP was observed in vitro than in vivo. No significant difference in ΔTIME was observed between test conditions. A significant, positive relationship was observed between radiant exposure and ΔTEMP for both conditions (in vivo: r2=0.917; p<0.001; in vitro: r2=0.919; p<0.001). Conclusion: Although the in vitro model overestimated in vivo PT increase, in vitro PT rise was close to in vivo values for clinically relevant exposure modes.

Evaluation of temperature increase during in-office bleaching

ABSTRACT The use of light sources in the bleaching process reduces the time required and promotes satisfactory results. However, these light sources can cause an increase in the pulp temperature. Objective The purpose of the present study was to measure the increase in intrapulpal temperature induced by different light-activated bleaching procedures with and without the use of a bleaching gel. Material and Methods A human maxillary central incisor was sectioned 2 mm below the cementoenamel junction. A K-type thermocouple probe was introduced into the pulp chamber. A 35% hydrogen peroxide bleaching gel was applied to the vestibular tooth surface. The light units used were a conventional halogen, a hybrid light (only LED and LED/Laser), a high intensity LED, and a green LED light. Temperature increase values were compared by two-way ANOVA and Tukey´s tests (p<0.05). Results There were statistically significant differences in temperature increases between the different light sources used and between the same light sources with and without the use of a bleaching gel. The presence of a bleaching gel generated an increase in intra-pulpal temperature in groups activated with halogen light, hybrid light, and high intensity LED. Compared to the other light sources, the conventional halogen lamp applied over the bleaching gel induced a significant increase in temperature (3.83±0.41°C). The green LED unit with and without gel application did not produce any significant intrapulpal temperature variations. Conclusion In the present study, the conventional halogen lamp caused the highest increase in intrapulpal temperature, and the green LED caused the least. There was an increase in temperature with all lights tested and the maximum temperature remained below the critical level (5.5°C). The addition of a bleaching gel led to a higher increase in intrapulpal temperatures.

Influência de diferentes densidades de energia do laser de baixa intensidade em fibroblastos derivados da polpa de dentes decíduos / Influence of low-level laser with different energy densities on pulp fibroblasts from human primary teeth

O objetivo deste trabalho foi comparar os efeitos de diferentes densidades de energia e irradiâncias do Laser de Baixa Intensidade (LBI), variando em função do tempo de irradiação e potência, na viabilidade e proliferação de fibroblastos derivados da polpa de dentes decíduos humanos (HPF). HPF foram cultivados em DMEM e usados entre a 4ª e 8ª passagem. Os grupos foram divididos de acordo com diferentes densidades de energia, variando: Tempo de irradiação - Grupo I Ia (1,2 J/cm2 - 5 mW - 10 s), Ib (2,5 J/cm2 - 5 mW - 20 s), Ic (3,7 J/cm2 - 5 mW - 30 s), Id (5,0 J/cm2 - 5 mW - 40 s), e Ie (6,2 J/cm2 - 5 mW - 50 s); ou potência - Grupo II IIa (1,2 J/cm2 - 5 mW - 10 s), IIb (2,5 J/cm2 - 10 mW - 10 s), IIc (3,7 J/cm2 - 15 mW - 10 s), IId (5,0 J/cm2 - 20 mW - 10 s), e IIe (6,2 J/cm2 - 25 mW - 10 s). Células não irradiadas - cultivadas em condições nutricionais regulares - 10% Soro Fetal Bovino (SFB) (If e IIf) e células não irradiadas - cultivadas em déficit nutricional - 1% SFB (Ig e IIg), foram consideradas controles positivos e negativos, respectivamente. A viabilidade e proliferação celular foram avaliadas, repesctivamente, pelas técnicas MTT e Cristal violeta (CV), nos períodos de 24, 48 e 72 horas após a irradiação. Os dados obtidos foram submetidos à análise estatística por ANOVA 2 critérios, seguido pelo teste de Tukey (P<0,05). No ensaio MTT, os controles negativos, Ig e IIg, apresentaram significativamente menor viabilidade em relação aos correspondentes grupos experimentais: IIa e IIb, 24 horas após a irradiação; Ia, Ib, Ie, If e IIf no período de 48 horas; e Ib-If, assim como, IIa-IIf após 72 horas. Nos diferentes períodos de avaliação do ensaio CV, todos os grupos, exceto Ie, IIe e If, exibiram significativamente maior proliferação em comparação aos respectivos controles negativos. Dentro de um mesmo grupo nos diferentes períodos, os grupos If e IIe apresentaram menor viabilidade durante o período de 24 horas em comparação ao período de 72 horas pelo ensaio MTT. Na avaliação intragrupos, o ensaio CV revelou menor proliferação no período de 24 horas em comparação aos períodos de 48 e 72 horas, independente do grupo avaliado. Os diferentes protocolos de irradiação, grupos I e II, não apresentaram diferença estatisticamente significativa na viabilidade e proliferação celular entre densidades de energia iguais com irradiâncias diferentes durante os períodos avaliados. De acordo com os resultados obtidos, as diferentes densidades de energia e irradiâncias propostas não prejudicaram a viabilidade e proliferação de fibroblastos pulpares de dentes decíduos humanos. A variação do protocolo de irradiação LBI, em função do tempo ou da potência, não interferiram nas respostas celulares após a aplicação da mesma densidade de energia com irradiâncias diferentes.(AU)

The aim of this study was to compare the effects of Low-level laser (LLL) with different energy densities and irradiances, varying according to the irradiation time and power, on cell viability and proliferation of pulp fibloblasts from human primary teeth (HPF). HPF were culture in DMEM and used between 4th and 8th passages. Groups were divided according to different energy densities, varying: Time of irradiation Ia (1.2 J/cm2 - 5 mW - 10 s), Ib (2.5 J/cm2 - 5 mW - 20 s), Ic (3.7 J/cm2 - 5 mW - 30 s), Id (5.0 J/cm2 - 5 mW - 40 s), and Ie (6.2 J/cm2 - 5 mW - 50 s); or output power - Grupo II IIa (1.2 J/cm2 - 5 mW - 10 s), IIb (2.5 J/cm2 - 10 mW - 10 s), IIc (3.7 J/cm2 - 15 mW - 10 s), IId (5.0 J/cm2 - 20 mW - 10 s), e IIe (6.2 J/cm2 - 25 mW - 10 s). Non-irradiated cells - grown in regular nutritional conditions - 10% Fetal Bovine Serum (FSB) (If and IIf) and non-irradiated cells - grown in nutritional deficit - 1% FBS (Ig and IIg) were considered positive and negative controls, respectively. Cell viability and proliferation were respectively assessed through MTT and Crystal violet (CV) assays at 24, 48 and 72h after irradiation. Data were submitted to statistical analysis by ANOVA 2 criteria, followed by Tukey test (P<0.05). In the MTT assay, the negative controls, Ig and IIg, showed significantly lower viability in relation to the corresponding groups: IIa and IIb 24 hours after irradiation; Ia, Ib, Ie, If and IIf at 48 hours period; and Ib-If, as IIa-IIf, after 72 hours. At different periods of evaluation of CV assay, all groups, except Ie, IIe and If, exhibited significantly higher proliferation compared to the respective negative controls. Within the same group at different periods, groups If and IIe showed lower viability during 24 hours compared to 72 hours period by MTT assay. In the intragroup evaluation, CV assay revealed lower proliferation at 24 hours compared to 48 and 72 hours periods, regardless of the evaluated group. Different irradiation protocols, groups I and II, showed no statistically significant differences on cell viability and proliferation among equals energy densities with different irradiances at the evaluated periods. According to these findings, different LLL energy densities and irradiances proposed did not impair viability and proliferation of pulp fibloblasts from human primary teeth. The variation of the LLL irradiation protocol, by the time or power, did not interfere in cellular responses after the application of the same energy density with different irradiances.(AU)

Low-intensity laser phototherapy enhances the proliferation of dental pulp stem cells under nutritional deficiency

Abstract Dental trauma in immature permanent teeth can damage pulp vascularization, which leads to necrosis and cessation of apexogenesis. Studies on tissue engineering using stem cells from human exfoliated deciduous teeth (SHEDs) have yielded promising results. Laser phototherapy (LPT) is able to influence the proliferation and differentiation of these cells, which could improve tissue engineering. SHEDs (eighth passage) were seeded into 96-well culture plates (103 cells/well) and were grown in culture medium supplemented with 15% defined fetal bovine serum (FBS) for 12 h. After determining the appropriate nutrition deficiency status (5% FBS), the cells were assigned into four groups: 1) G1 - 15% FBS (positive control); 2) G2 - 5% FBS (negative control); 3) G3 - 5% FBS+LPT 3 J/cm2; and 4) G4 - 5% FBS+LPT 5 J/cm2. For the LPT groups, two laser irradiations at 6 h intervals were performed using a continuous wave InGaAlP diode laser (660 nm, with a spot size of 0.028 cm2, 10 mW) in punctual and contact mode. Cell viability was assessed via an MTT reduction assay immediately after the second laser irradiation (0 h) and 24, 48, and 72 h later. We found that G3 and G4 presented a significantly higher cell growth rate when compared with G2 (p < 0.01). Moreover, G4 exhibited a similar cell growth rate as G1 throughout the entire experiment (p > 0.05). These findings indicate that LPT with 5 J/cm2 can enhance the growth of SHEDs during situations of nutritional deficiency. Therefore, LPT could be a valuable adjunct treatment in tissue engineering when using stem cells derived from the dental pulp of primary teeth.

Efeito do laser de baixa potência sobre células tronco de polpa de dentes decíduos esfoliados humanos (SHED) / Effect of low level laser on stem cells from human exfoliated deciduous teeth (SHED)

Avaliou-se a proliferação das células tronco da polpa de dentes decíduos esfoliados humanos (SHED) após aplicação única do laser de baixa potência. Foi realizada a análise da viabilidade das SHED cultivadas sob déficit nutricional e em condições ideais após irradiação com o laser de baixa potência vermelho de Indio Gálio Alumínio e Fósforo - InGaAlP (660nm, 40mW e 10J/cm2) e infravermelho (780nm, 40mW e 10J/cm2) durante 4 e 8 segundos, nos períodos de 24, 48 e 72 horas através dos ensaios de redução do MTT e do ensaio colorimétrico de Busatti e Gomes. Para análise estatística utilizou-se o teste ANOVA complementado pelo teste de Tukey com nível de significância de 5% (p< 0,05). Observou-se tanto com o MTT quanto com o ensaio colorimétrico de Busatti e Gomes uma tendência de aumento da proliferação celular diretamente relacionada à dose do LBP, estatisticamente significante nos períodos de 24, 48 e 72 horas. Ao analisar os resultados e considerando os parâmetros utilizados e o protocolo de LBP, pode-se concluir que o LBP promoveu a proliferação das SHED tanto a 660 nm quanto a 780nm, pode influenciar a viabilidade e a proliferação das SHED nas doses e comprimentos de onda utilizados e os ensaios do MTT e colorimétrico de Busati e Gomes demonstraram dentro de suas limitações ser eficientes para determinar a viabilidade e proliferação das SHED...

It was evaluated the proliferation of stem cells from human exfoliated deciduous teeth (SHED) after a single application of low power laser. The viability of SHED grown under ideal conditions and under nutritional deficit after irradiation with red laser (660/780nm, 10J/cm2 and 40mW) during periods of 4 and 8 seconds was analyzed through the MTT reduction assays and rapid colorimetric assay of Busatti and Gomes. Statistical analysis was performed using the ANOVA and Tukey´s multiple comparisons test with a significance level of 5% (p < 0.05). It was observed with the MTT assay and Busatti and Gomes assay a trend of cell proliferation increase directly releated to the irradiation dose, statistically significant. After 24, 48 and 72 hours, all the groups showed higher cell proliferation when compared to control. Analyzing the results and considering the used parameters and LBP protocol, it can be concluded that LBP promoted the proliferation of SHED both 660nm and 780nm according to the dosage and wavelengths used, and MTT assay and colorimetric Busatti and Gomes demonstrated within their limitations to be effective in determining the viability and proliferation of SHED...

Efeito do laser de baixa potência sobre células tronco de polpa de dentes decíduos esfoliados humanos (SHED) / Effect of low level laser on stem cells from human exfoliated deciduous teeth (SHED)

Avaliou-se a proliferação das células tronco da polpa de dentes decíduos esfoliados humanos (SHED) após aplicação única do laser de baixa potência. Foi realizada a análise da viabilidade das SHED cultivadas sob déficit nutricional e em condições ideais após irradiação com o laser de baixa potência vermelho de Indio Gálio Alumínio e Fósforo - InGaAlP (660nm, 40mW e 10J/cm2) e infravermelho (780nm, 40mW e 10J/cm2) durante 4 e 8 segundos, nos períodos de 24, 48 e 72 horas através dos ensaios de redução do MTT e do ensaio colorimétrico de Busatti e Gomes. Para análise estatística utilizou-se o teste ANOVA complementado pelo teste de Tukey com nível de significância de 5% (p< 0,05). Observou-se tanto com o MTT quanto com o ensaio colorimétrico de Busatti e Gomes uma tendência de aumento da proliferação celular diretamente relacionada à dose do LBP, estatisticamente significante nos períodos de 24, 48 e 72 horas. Ao analisar os resultados e considerando os parâmetros utilizados e o protocolo de LBP, pode-se concluir que o LBP promoveu a proliferação das SHED tanto a 660 nm quanto a 780nm, pode influenciar a viabilidade e a proliferação das SHED nas doses e comprimentos de onda utilizados e os ensaios do MTT e colorimétrico de Busati e Gomes demonstraram dentro de suas limitações ser eficientes para determinar a viabilidade e proliferação das SHED.

It was evaluated the proliferation of stem cells from human exfoliated deciduous teeth (SHED) after a single application of low power laser. The viability of SHED grown under ideal conditions and under nutritional deficit after irradiation with red laser (660/780nm, 10J/cm2 and 40mW) during periods of 4 and 8 seconds was analyzed through the MTT reduction assays and rapid colorimetric assay of Busatti and Gomes. Statistical analysis was performed using the ANOVA and Tukey´s multiple comparisons test with a significance level of 5% (p < 0.05). It was observed with the MTT assay and Busatti and Gomes assay a trend of cell proliferation increase directly releated to the irradiation dose, statistically significant. After 24, 48 and 72 hours, all the groups showed higher cell proliferation when compared to control. Analyzing the results and considering the used parameters and LBP protocol, it can be concluded that LBP promoted the proliferation of SHED both 660nm and 780nm according to the dosage and wavelengths used, and MTT assay and colorimetric Busatti and Gomes demonstrated within their limitations to be effective in determining the viability and proliferation of SHED.

In vitro effect of low intensity laser on the cytotoxicity produced by substances released by bleaching gel

This in vitro study aimed to analyze the effect of different parameters of phototherapy with low intensity laser on the viability of human dental pulp fibroblasts under the effect of substances released by bleaching gel. Cells were seeded into 96 wells plates (1 x 10³ cells/well) and placed in contact with culture medium conditioned by a 35 percent hydrogen peroxide bleaching gel for 40 minutes, simulating the clinical condition of the in-office bleaching treatment. Cells cultured in ideal growth conditions served as positive control group (PC), and the cells grown in conditioned medium and non-irradiated served as negative control group (NC). Cells grown in conditioned medium were submitted to a single irradiation with a diode laser (40 mW, 0.04 cm²) emitting at visible red (660 nm; RL) or near infrared (780 nm; NIR) using punctual technique, in contact mode and energy densities of 4, 6 or 10 J/cm². The cell viability was analyzed through the MTT reduction assay immediately and 24 hours after the irradiation. The data was compared by ANOVA followed by the Tukey's test (p < 0.05). The cell viability increased significantly in 24 hours within each group. The PC presented cell viability significantly higher than NC in both experimental times. Only the NIR/10 J/cm² group presented cell viability similar to that of PC in 24 hours. The phototherapy with low intensity laser in defined parameters is able to compensate the cytotoxic effects of substances released by 35 percent hydrogen peroxide bleaching gel.

invitro study on the temperature changes of dentin, irradiated by CO2 and Er: Cr;YSGG laser

Aim of this in-vitro study was the evaluation of temperature changes due to irradiation of two different lasers used for the reduction of dentinal hypersensitivity and their effect on the pulp damage. The study was done for two dentin thicknesses. Twenty intact extracted third molars were prepared by longitudinal ground sectioning for 1 and 2 mm dentin thicknesses while a thermocouple was positioned at the inner surface of the dentin disk. Thermal evaluation was assessed by a KJT digital thermometer. During the test, the data produced by the thermometer was transferred and logged into a PC via RS232 serial port. CO2 laser [Ultra pulse, 50W, 100?sec, Spot size: 0.5 mm] and Er,Cr;YSGG laser [Free-running pulse mode,0. 25W, 140?sec, 12.50 milli-joules] irradiations were randomly performed upon the dentin surfaces. The collected data was analyzed by two-way ANOVA test. The mean temperature rise in 1mm dentinal thickness was 8.57°C which was significantly higher than 3.63°C in 2mm dentin thickness [P<0.001] and higher than the threshold temperature for pulp damage; however, no significant difference was noted between the two lasers [P=0.355]. After removing the CO2 laser, the temperature decreased to the initial level faster than the time needed for Er,Cr;YSGG laser [44.47°C versus 62.82°C][P<0.001]. In other words, in both lasers the temperature decrease in 2mm dentinal disc was faster than 1mm dentinal disc. The temperature rise due to both lasers for 1mm of dentinal thickness was in excess of safe limit for the tissue and it would probably result in pulpal damage. In the case of 2mm dentinal thickness, the temperature rise was not higher than the safe limit and it would not damage the pulp in clinical conditions

Intrapupal temperature variation during Er,Cr: YSGG enamel irradiation on carries prevention

Studies have shown the cariostatic effect of Er,Cr:YSGG (2.78 mm) laser irradiation on human enamel and have suggested its use on caries prevention. However there are still no reports on the intrapulpal temperature increase during enamel irradiation using parameters for caries prevention. The aim of this in vitro study was to evaluate the temperature variation in the pulp chamber during human enamel irradiation with Er,Cr:YSGG laser at different energy densities. Fifteen enamel blocks obtained from third molars (3 x 3 x 3 mm) were randomly assigned to 3 groups (n=5): G1 - Er,Cr:YSGG laser 0.25 W, 20 Hz, 2.84 J/cm², G2 - Er,Cr:YSGG laser 0.50 W, 20 Hz, 5.68 J/cm², G3 - Er,Cr:YSGG laser 0.75 W, 20 Hz, 8.52 J/cm². During enamel irradiation, two thermocouples were fixed in the inner surface of the specimens and a thermal conducting paste was used. One-way ANOVA did not show statistically significant difference among the experimental groups (a=0.05). There was intrapulpal temperature variation <0.1ºC for all irradiation parameters. In conclusion, under the tested conditions, the use of Er,Cr:YSGG laser with parameters set for caries prevention lead to an acceptable temperature increase in the pulp chamber.

Estudo da resposta pulpar em pacientes portadores de neoplasias malignas de cabeça e pescoço submetidos à radioterapia / Evaluation of pulp response in patients undergone radiation therapy for treatment of head and neck cancer

Introdução: a radioterapia promove alterações na cavidade oral e na polpa dental que conduzem a mudanças na sensibilidade frente a estímulos em contato com a superfície dental. Torna-se necessário investigar a ocorrência dessas alterações e estabelecer o diagnóstico pulpar e perirradicular dos elementos remanescentes em áreas submetidas à irradiação ionizante, com o intuito de prevenirem-se os processos patológicos periapicais, descritos como fatores precipitadores de osteorradionecrose. Objetivo: avaliar as respostas pulpares ao teste de vitalidade com gás refrigerante tetrafluoroetano, em pacientes, submetidos à radioterapia e em um grupo de controle, em três agrupamentos dentários distintos (incisivos inferiores, incisivos superiores e caninos). Casuística e método: foram testados 91 dentes hígidos de pacientes de ambos os gêneros submetidos à radioterapia para o tratamento de neoplasias malignas de cabeça e pescoço e 103 dentes de pacientes do grupo de controle. Resultados: verificou-se que a diferença significativa (p<.05) entre os dois grupos com relação às respostas negativas apresentadas; não houve diferença entre os agrupamentos dentários avaliados. Conclusão: os pacientes submetidos à radioterapia de cabeça e pescoço apresentam amior número de elementos dentais com resposta negativa ao teste de vitalidade pulpar quando comparados com um grupo de controle

Introduction: alterations promoted by radiation therapy in the oral cavity and in dental pulp leads to sensitiveness changes in enamel surface when in contact with some external stimuli. The investigation of these changes and the establishment of the pulpal and perapicaql diagnosis of the elements remained in irradiated sites, in order to prevent pathological process that have been described as osteoradionecrosis developing factors, is mandatory. Objective: to evaluate the pulp response obtained by tetrafluoroethane in patients under gone radiation therapy and in a control group, in three dental groups (maxillary incisors, mandibular incisors and canines). Patients and methods: 91 teeth were tested in patients in both genders submitted to radiation therapy for head and neck malignances and 103 teeth in a control group. Results: the results showed sifnificant difference (p<.05) between the groups evaluated related to negative responses, despite no difference between the dental groups. Conclusion: patients undergoing radiation therapy in tghe head and neck present a greater number of teeth with negative response to the vitality test in comparison to the control group

El láser y la Odontopediatría / Laser and Pediatric Dentistry

Aunque la energís láser se conoce desde hace 40 años, los últimos 10 años ha crecido su importancia en la práctica odontológica, hoy en día existen diferentes tipos de energía láser, diferentes materiales láser, que nos permiten encarar tanto la prevención como el diagnóstico y el tratamiento de una gran parte de las patologías bucales. Con el láser puede aumentarse la resistencia del esmalte al avance del proceso de la caries, pueden diagnosticarse incipientes pérdidas de sustancia inorgánica en la superficie del esmalte o prepararse cavidades con destino a la operatoria adhesiva, sin dolor y sin anestesia en la mayoría de los casos. También hay láseres que nos permiten realizar exitosos procedimientos quirúrgicos en los tejidos blandos otratamientos médicos con efectos antiinflamatorios, analgésicos, antiedematosos o cicatrices

Efectos de la remoción químico-mecánica (GK-10/E) sobre la pulpa dental / Chemo-mechanical removing effects (GK-10/E) over dental pulp

El objetivo del presente trabajo fue determinar la efectividad y la veracidad de todas las investigaciones realizadas con el Caridex, a objeto de ser recomendado y utilizado en servicios de atención masiva, y tratar de simplificar los procedimientos de eliminación de caries tanto en los servicios públicos como privados..

Determinación de la longitud de trabajo de los conductos radiculares a través de métodos electrónicos / Electronic determination of root canal length work

Se realiza una revisión bibliográfica, que permite comparar, a través de los resultados la efectividad de la conductometria electrónica con la obtenida por métodos convencionales dando a conocer resultados obtenidos, confiabilidad y establecer ventajas y desventajas de este método..