RESUMO
Background: This investigation set out to compare the impacts of low-level diode laser (LLDL) and red light-emitting diode (LED) on the survival of human dental pulp stem cells (hDPSCs) and osteogenic/odontogenic differentiation. Methods and materials: In this ex vivo experimental study, the experimental groups underwent the irradiation of LLDL (4 J/cm2 energy density) and red LED in the osteogenic medium. Survival of hDPSCs was assessed after 24 and 48 h (n = 9) using the methyl thiazolyl tetrazolium (MTT) assay. The assessment of osteogenic/odontogenic differentiation was conducted using alizarin red staining (ARS; three repetitions). The investigation of osteogenic and odontogenic gene expression was performed at two time points, specifically 24 and 48 h (n = 12). This analysis was performed utilizing real-time reverse-transcription polymerase chain reaction (RT-PCR). The groups were compared at each time point using SPSS version 24. To analyze the data, the Mann-Whitney U test, analysis of variance, Tukey's test, and t-test were utilized. Results: The MTT assay showed that LLDL significantly decreased the survival of hDPSCs after 48 h, compared with other groups (p < 0.05). The qualitative results of ARS revealed that LLDL and red LED increased the osteogenic differentiation of hDPSCs. LLDL and red LED both upregulated the expression of osteogenic/odontogenic genes, including bone sialoprotein (BSP), alkaline phosphatase (ALP), dentin matrix protein 1 (DMP1), and dentin sialophosphoprotein (DSPP), in hDPSCs. The LLDL group exhibited a higher level of gene upregulation (p < 0.0001). Conclusions: The cell survival of hDPSCs was reduced, despite an increase in osteogenic/odontogenic activity. Clinical relevance: Introduction of noninvasive methods in regenerative endodontic treatments.
Assuntos
Diferenciação Celular , Sobrevivência Celular , Polpa Dentária , Lasers Semicondutores , Terapia com Luz de Baixa Intensidade , Odontogênese , Osteogênese , Células-Tronco , Humanos , Polpa Dentária/citologia , Polpa Dentária/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Osteogênese/efeitos da radiação , Células-Tronco/efeitos da radiação , Células-Tronco/citologia , Sobrevivência Celular/efeitos da radiação , Odontogênese/efeitos da radiação , Células Cultivadas , Luz VermelhaRESUMO
To investigate the effects of gallium-aluminum-arsenide (GaAlAs) diode laser low-level laser therapy (LLLT) on angiogenesis and dentinogenesis of the dentin-pulp complex in a human tooth slice-based in vitro model. Forty tooth slices were prepared from 31 human third molars. Slices were cultured at 37 °C, 5% CO2, and 95% humidity and randomly assigned to one of the following groups: group I: no laser treatment, group II: 660-nm diode laser; energy density = 1 J/cm2, group III: 660-nm diode laser; energy density = 3 J/cm2, group IV: 810-nm diode laser; energy density = 1 J/cm2 and group V: 810-nm diode laser; energy density = 3 J/cm2. LLLT was applied on the third and fifth days of culture. After 7 days, tissues were retrieved for real-time RT-PCR analysis to investigate the expression of VEGF, VEGFR2, DSPP, DMP-1, and BSP in respect to controls. Lower energy density (1 J/cm2) with the 660 nm wavelength showed a statistically significant up-regulation of both angiogenic (VEGF: 15.3-folds and VEGFR2: 3.8-folds) and odontogenic genes (DSPP: 6.1-folds, DMP-1: 3-fold, and BSP: 6.7-folds). While the higher energy density (3 J/cm2) with the 810 nm wavelength resulted in statistically significant up-regulation of odontogenic genes (DSPP: 2.5-folds, DMP-1: 17.7-folds, and BSP: 7.1-folds), however, the angiogenic genes had variable results where VEGF was up-regulated while VEGFR2 was down-regulated. Low-level laser therapy could be a useful tool to promote angiogenesis and dentinogenesis of the dentin-pulp complex when parameters are optimized.
Assuntos
Técnicas de Cultura de Células , Polpa Dentária/efeitos da radiação , Dentinogênese/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Adulto , Feminino , Humanos , Terapia com Luz de Baixa Intensidade/métodos , Masculino , Neovascularização Fisiológica/efeitos da radiação , Odontogênese/efeitos da radiação , Adulto JovemRESUMO
Several studies suggest a biomodulatory influence of low-level laser irradiation in the inflammatory and reparative processes of dental tissues. The aim of this study was to investigate the stimulatory effect of 808-nm laser irradiation on root development of rat molars and also to evaluate the histological reaction of pulp and periapical tissues. Twenty-four 30-day-old Wistar male rats were randomly assigned to three-time and five-time laser therapy groups. After initial x-ray, using mammography equipment, laser energy was applied at a wavelength of 808 nm (2 J/cm(2), 100 mW, 20 s) to the midroot area of the lower molars of one side of mouth at repeated intervals of the 48 h. The animals were killed 1 day after the final treatment, and root length development of the experimental samples was compared to contra-lateral non-irradiated molars using mammography. The histological reaction of the pulp and periapical tissue was evaluated under light microscopy. Root development was more advanced in irradiated groups than in the non-irradiated controls (p < 0.001). No significant differences, however, could be found between the root development changes in the three-time and five-time laser therapy groups (p > 0.05). Histological findings showed that the occurrence of secondary cement formation was significantly higher in the irradiation groups compared to the controls (p = 0.003). However, there were no statistically significant differences for the frequencies of pulp hyperemia, periodontal ligament fiber organization, or lamina dura remodeling between the groups (p > 0.05). Under the conditions used in this study, 808-nm low-level laser accelerates the rat molar root development in the presence of favorable histological reactions.
Assuntos
Terapia com Luz de Baixa Intensidade , Dente Molar/efeitos da radiação , Raiz Dentária/efeitos da radiação , Animais , Polpa Dentária/efeitos da radiação , Masculino , Modelos Animais , Dente Molar/diagnóstico por imagem , Odontogênese/efeitos da radiação , Tecido Periapical/efeitos da radiação , Radiografia , Ratos , Ratos Wistar , Raiz Dentária/diagnóstico por imagem , Raiz Dentária/crescimento & desenvolvimentoRESUMO
BACKGROUND AND OBJECTIVES: Third molars (wisdom teeth) in humans have limited functional value and cause a high frequency of morbidity. A window of opportunity exists to stop their development. The purpose of this study is to evaluate whether a long pulse diode laser can selectively prevent third molar development in a rat animal model. STUDY DESIGN/MATERIALS AND METHODS: The study calculated theoretical diode laser parameters and irradiated 2- to 4-day-old rat pups in the posterior aspects of the oral cavity near the time of tooth bud initiation. Rats were sacrificed at 35-50 days and examined intraorally and radiographically for third molar development. Control and treated sides were compared histologically. RESULTS: Nearly half of the maxillary third molars and 80% of the mandibular third molars were selectively prevented from developing using a 20 W diode laser with infrequent, minor adverse effects on maxillary jaw growth. CONCLUSIONS: This study confirms the hypothesis that controlled, non-invasive laser treatment delivered on the surface of the oral mucosa near the site of tooth bud formation can selectively achieve third molar agenesis. It also offers hope that further innovative studies may bring about a safe, non-invasive laser treatment to selectively prevent third molar development in humans.