Effects of diode laser irradiation on dental pulps in rats.

OBJECTIVES: To evaluate the effects of different power densities of diode laser on dental pulps in rats. BACKGROUND: In this study, we used the maxillary central incisors (n=80) of the 40 adult male Wistar albino rats. METHODS: Rats were randomly divided into four groups according to power densities of diode laser (n=10). Histopathological changes in pulp and height of odontoblast layer were examined . All data were compared statistically using Mann‒Whitney U (Bonferroni) test, p<0.05. RESULTS: G2 displayed slight histolopathologic alterations such as odontoblast cell disorganization and irregularities in cell extensions. Alterations were more prominent in the G3 than G2. Although the lowest odontoblast layer was measured in the G4, the difference in height of odontoblast layer among the groups was not found to be statistically significant. CONCLUSION: It was concluded that the use of diode laser caused changes at the cellular level in histological examination and may induce the formation of tertiary dentin by influencing the secretory activity of odontoblasts. As long as used in accordance with the recommended procedure, the diode laser can be safely used in dental hard tissues (Tab. 1, Fig. 4, Ref. 15).

Effects of Er:YAG and Diode Laser Irradiation on Dental Pulp Cells and Tissues.

Vital pulp therapy (VPT) is to preserve the nerve and maintain healthy dental pulp tissue. Laser irradiation (LI) is beneficial for VPT. Understanding how LI affects dental pulp cells and tissues is necessary to elucidate the mechanism of reparative dentin and dentin regeneration. Here, we show how Er:YAG-LI and diode-LI modulated cell proliferation, apoptosis, gene expression, protease activation, and mineralization induction in dental pulp cells and tissues using cell culture, immunohistochemical, genetic, and protein analysis techniques. Both LIs promoted proliferation in porcine dental pulp-derived cell lines (PPU-7), although the cell growth rate between the LIs was different. In addition to proliferation, both LIs also caused apoptosis; however, the apoptotic index for Er:YAG-LI was higher than that for diode-LI. The mRNA level of odontoblastic gene markers-two dentin sialophosphoprotein splicing variants and matrix metalloprotease (MMP)20 were enhanced by diode-LI, whereas MMP2 was increased by Er:YAG-LI. Both LIs enhanced alkaline phosphatase activity, suggesting that they may help induce PPU-7 differentiation into odontoblast-like cells. In terms of mineralization induction, the LIs were not significantly different, although their cell reactivity was likely different. Both LIs activated four MMPs in porcine dental pulp tissues. We helped elucidate how reparative dentin is formed during laser treatments.

Photopolymerization of cell-laden gelatin methacryloyl hydrogels using a dental curing light for regenerative dentistry.

Photopolymerized hydrogels, such as gelatin methacryloyl (GelMA), have desirable biological and mechanical characteristics for a range of tissue engineering applications. OBJECTIVE: This study aimed to optimize a new method to photopolymerize GelMA using a dental curing light (DL). METHODS: Lithium acylphosphinate photo-initiator (LAP, 0.05, 0.067, 0.1% w/v) was evaluated for its ability to polymerize GelMA hydrogel precursors (10% w/v) encapsulated with odontoblast-like cells (OD21). Different irradiances (1650, 2300 and 3700mW/cm2) and photo-curing times (5-20s) were tested, and compared against the parameters typically used in UV light photopolymerization (45mW/cm2, 0.1% w/v Irgacure 2959 as photoinitiator). Physical and mechanical properties of the photopolymerized GelMA hydrogels were determined. Cell viability was assessed using a live and dead assay kit. RESULTS: Comparing DL and UV polymerization methods, the DL method photopolymerized GelMA precursor faster and presented larger pore size than the UV polymerization method. The live and dead assay showed more than 80% of cells were viable when hydrogels were photopolymerized with the different DL irradiances. However, the cell viability decreased when the exposure time was increased to 20s using the 1650mW/cm2 intensity, and when the LAP concentration was increased from 0.05 to 0.1%. Both DL and UV photocrosslinked hydrogels supported a high percentage of cell viability and enabled fabrication of micropatterns using a photolithography microfabrication technique. SIGNIFICANCE: The proposed method to photopolymerize GelMA cell-laden hydrogels using a dental curing light is effective and represents an important step towards the establishment of chair-side procedures in regenerative dentistry.

"Metabolism of Odontoblast-like cells submitted to transdentinal irradiation with blue and red LED".

OBJECTIVES: The present study evaluated the trans-dentinal effect of light emitting diodes (LEDs) irradiation on the metabolism of odontoblast-like cells. METHODS: Seventy-two dentin discs (0.2mm thick) were obtained from human molar teeth. MDPC-23 cells (20,000 cells/disc) were seeded on the pulpal side of the discs using DMEM, supplemented with 10% fetal bovine serum (FBS). After 12h, the culture medium was replaced with DMEM containing 0.5% FBS. After additional 12h, blue (455±10nm) or red (630±10nm) LEDs were used at irradiances of 80 and 40mW/cm2, respectively, to irradiate the occlusal side of the discs. The energy doses were fixed at 2 or 4J/cm2. Cell viability, alkaline phosphatase activity (ALP), total protein production and collagen synthesis were evaluated 72h after irradiation. Data were submitted to Kruskal-Wallis and Mann-Whitney tests (α=0.05). RESULTS: Red light promoted proliferative effects at the energy dose of 4J/cm2. Conversely, cell cultures irradiated with 2J/cm2 emitted by the blue light showed reduced viability. ALP production was stimulated by red light in comparison with blue light at 4J/cm2. Total protein production was reduced after exposure to blue light at 4J/cm2, while no effect was observed on collagen production. CONCLUSIONS: Irradiation with red LED at 4J/cm2 bio-stimulated the viability of odontoblast-like cells, whilst blue light had unfavorable effects on the cellular metabolism.

Effect of low-level laser therapy (λ780 nm) on the mechanically damaged dentin-pulp complex in a model of extrusive luxation in rat incisors.

In order to regenerate the dental pulp, many strategies have been developed as phototherapy. In the pulp repair, we do not know if gallium-aluminum-arsenide (GaAlAs) laser preserves the primary odontoblasts or stimulates the formation of more dentin matrix when dental pulp is damaged. The aim of the present study was to examine the effect of laser phototherapy (λ780 nm) on vascularization, inflammation, density of the primary odontoblast layer, and formation of reactionary and reparative dentin in the dental pulp by provoking extrusion of the rat incisor. The upper incisors were extruded 3 mm and then repositioned into their original sockets followed by a laser irradiation of the palatal mucosa (λ = 780 nm; p = 70 mW; CW; 4.2 J/cm2; 60 s) every 48 h. Non-traumatized and/or non-irradiated incisors were used as the controls. At 8 and 30 days after surgery, incisors were processed for histological and histomorphometric analysis. Morphological analysis revealed no differences in vascularization between groups, but showed discrete inflammation in some non-irradiated and injured specimens, which correlated with a more irregular reparative dentin. The density of primary odontoblasts in the groups treated with lasers was higher when compared to non-irradiated groups, but no statistically significant difference between groups (p > 0.05). The thickness of the tertiary dentin was increased in both traumatized groups with no statistically significant difference between non-irradiated and irradiated groups (p > 0.05).The present findings revealed that the GaAlAs laser induced small changes on dentin-pulp complex, with more regular dentin matrix in the irradiated dental pulps.

Near-infrared laser increases MDPC-23 odontoblast-like cells proliferation by activating redox sensitive pathways.

Near infrared laser is known to induce biostimulatory effects, resulting in cell proliferation enhancement. Although such positive effect is widely exploited in various clinical applications, molecular mechanisms involved are still poorly understood. The aim of the study was to investigate the ability of laser stimulation to increase cell proliferation through an early activation of three redox sensitive pathways, namely Nrf-2, NF-κB and ERK in a rat odontoblast-like cell line (MDPC-23 cells). MDPC-23 cells were irradiated with different energy settings (0-50J, corresponding to 0-32.47J/cm2) and cell proliferation was evaluated by cell counting. Nrf-2, NF-κB and ERK signaling pathways activation was investigated through Western blot analysis. Our results show that a single 25J laser stimulation is able to increase cell proliferation and that this effect could be increased by repeating the stimulation twice with a time lapse of 24h. Western blot experiments demonstrated that laser stimulation is able to induce an early activation response in intracellular signaling, with an overlapping time pattern between the three considered pathways. Results discussed in this paper reveal a complex mechanism underlying near-infrared induced increase in pre-odontoblasts proliferation, involving three survival pathways that can act both separately or through reciprocal crosstalk. In particular, data presented suggest an important role for ERK pathway that could act directly by stimulating cell proliferation but can also induce both Nrf-2 and NF-κB activation, acting as a critical cellular checkpoint in response to imbalanced redox state generated by a laser induced increase in ROS production.

Pre-odontoblast proliferation induced by near-infrared laser stimulation.

OBJECTIVE: Laser therapy is known to stimulate cell proliferation and differentiation, an effect called "biostimulation". Although many clinical applications of laser therapy take advantage from such positive effect, the underlying molecular mechanisms are not fully understood. The aim of this work was to investigate the effect of near-infrared laser stimulation on rat pre-odontoblast cells (MDPC-23 cells) and the molecular mechanism/s involved. MATERIALS AND METHODS: MDPC-23 cells were stimulated with a near-infrared (980 nm) laser source with different energy settings (1-50 J, corresponding to 0.65-32.47 J/cm2) and cell proliferation was evaluated by manual count. ERK 1/2 pathway activation was evaluated by Western blot analysis. RESULTS: 1-10 J stimulation (corresponding to 0.65-6.5 J/cm2) significantly increase MDPC-23 cell proliferation and such effect seems to be mediated by ERK 1/2 signalling pathway activation, showing a key role of ERK 1/2 pathway in mediating the proliferative response induced by laser stimulation. CONCLUSIONS: Near infrared laser stimulation with low energies (1-10 J) is able to increase cell proliferation through ERK 1/2 signalling pathway activation. At the same time, higher energy stimulation (25-50 J) induces an initial toxic effect, probably activating pro-apoptotic signalling molecules, downstream ERK 1/2 kinase. Such results foster the application of this therapeutic approach in different clinical settings in which a regenerative tissue response is needed.

Effects of CO2 Lasers on Dental Pulp Biology in Rats.

OBJECTIVE: The purpose of this study was to investigate the effects of CO2 lasers on the proliferation and differentiation of dental pulp cells, and their latent self-recovery in connection with their stemness using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. MATERIALS AND METHODS: The first molars from male Sprague-Dawley rats, each weighing ∼150-200 g, were used for this study. The upper first molars were irradiated with a 10,600 nm wavelength CO2 laser under identical parameters (2 W CO2 laser, energy 4J, energy density 203.84 J/cm(2) for 8.8 sec) through the dentin of the occlusal surface. The molars were extracted immediately, or at 1, 3 or 5 days after the laser irradiation. RT-PCR analysis using primers specific for heat shock protein 70 (Hsp70), adenosine triphosphate (ATP)-binding cassette transporter G2 (ABCG2), dentin sialophosphoprotein (DSPP), and dentin matrix protein 1 (DMP1), and immunohistochemistry using antibodies specific for proliferating cell nuclear antigen (PCNA), ABCG2, CD34, and CD44 were performed. RESULTS: RT-PCR analysis revealed that Hsp70 mRNA expression in the immediate group and ABCG2 mRNA expression at day 1 were the highest. DSPP and DMP1 mRNA expression in the laser-irradiated groups increased gradually, reaching its peak on the 5th day of the experiment, although no significant difference found among groups with regard to DMP1 expression. Immunohistochemically, PCNA-positive cells were observed at all times after the laser irradiation; however, they were most evident on day 3. CD44-positive cells were observed strongly on day 1 and day 3, while ABCG2-positive cells were the most evident on day 3. CONCLUSIONS: These results demonstrate that CO2 laser irradiation induces degeneration in the pulp tissue, which is then repaired by newly formed odontoblast-like cells.

Synthesis of dental matrix proteins and viability of odontoblast-like cells irradiated with blue LED.

To evaluate the effect of irradiation with light-emitting diode (LED; 455 nm) on the viability and synthesis of dentin matrix proteins by odontoblast-like cells, MDPC-23 cells were cultivated (10(4) cells/cm(2)) in 24-well culture plates. After 12 h incubation in Dulbecco's modified Eagle's medium (DMEM), the cells were submitted to nutritional restriction by means of reducing the concentration of fetal bovine serum (FBS) for an additional 12 h. Cells were irradiated one single time with one of the following energy densities (EDs): 0.5, 2, 4, 10, or 15 J/cm(2) and irradiance fixed at 20 mW/cm(2). Non-irradiated cells served as control. After 72 h, cells were evaluated with regard to viability (methylthiazol tetrazolium technique (MTT)), mineralization nodule (MN) formation, total protein (TP) production, alkaline phosphatase activity (ALP), and collagen synthesis (Sircol), n = 8. The data were submitted to Kruskal-Wallis and Mann-Whitney tests (p > 0.05). There was no statistical difference between the viability of cells irradiated or not (control), for all the EDs. However, an increase in TP was observed for all the EDs when compared with the control group. A reduced ALP activity was seen in all irradiated groups, except for the ED of 0.5 J/cm(2), which did not differ from the control. There was no difference between the irradiated groups and control regarding collagen synthesis, with the exception of the ED of 10 J/cm(2), which inhibited this cell function. Significant reduction in MN occurred only for the EDs of 0.5 and 2 J/cm(2). The single irradiation with blue LED (455 nm), irradiance of 20 mW/cm(2), and energy densities ranging from 0.5 to 15 J/cm(2) exerted no effective biostimulatory capacity on odontoblast-like cells.

GaAlAs laser-induced pulp mineralization involves dentin matrix protein 1 and osteopontin expression.

OBJECTIVES: GaAlAs lasers induce pulp mineralization by promoting reparative dentinogenesis. This study analyzed the expression of dentin matrix protein 1 (DMP1) and osteopontin in GaAlAs laser-irradiated rat molars, to examine the hypothesis that these proteins play a role in the laser-induced reparative dentinogenic process. MATERIALS AND METHODS: The mesial surfaces of the upper first molars of 8-week-old Wistar rats were irradiated with a pulsed GaAlAs laser. After 1-14 days, mRNA expression of DMP1 and osteopontin in the coronal pulp was analyzed using real-time PCR. DMP1, osteopontin, and heat shock protein 25 (HSP25) were immunolocalized at 1-21 days. RESULTS: The pulp exhibited a degenerative zone in its mesial portion on days 1-3, and progressive formation of reparative dentin lined with HSP25-immunoreactive odontoblast-like cells, from day 7 onwards. DMP1 and osteopontin mRNA expression were significantly upregulated on days 1-7 and 3-7, respectively. From day 7 onwards, DMP1 and osteopontin immunoreactivity colocalized along the boundary between the primary and reparative dentin. CONCLUSION: GaAlAs laser irradiation of rat molars induced upregulated DMP1 and osteopontin mRNA expression in the coronal pulp, followed by the formation of reparative dentin and the colocalization of DMP1 and osteopontin immunoreactivity at the site at which this tissue first appeared.

Metabolic activity of odontoblast-like cells irradiated with blue LED (455 nm).

Blue light emitting diodes (LEDs) are frequently used in dentistry for light activation of resin-based materials; however, their photobiostimulatory effects have not yet been fully investigated. This study aimed to investigate the effect of blue LED (455 nm) on the metabolism of odontoblast-like cells MDPC-23. Energy doses of 2 and 4 J/cm(2) were used at 20 mW/cm(2) fixed power density. MDPC-23 cells were seeded at 10,000 cells/cm(2) density in Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum (FBS). After 12 h, the culture medium was replaced with new DMEM supplemented with 0.5 % of FBS, and the cells were incubated for further 12 h. After that, single irradiation was performed to the culture, under selected parameters. Cell viability evaluations (Alamar Blue Assay, n = 12), number of viable cells (Trypan Blue Assay, n = 12), morphological analysis by scanning electron microscopy (SEM, n = 2), gene expression (n = 6) of alkaline phosphatase (Alp), collagen (Col-1a1), and dental matrix protein (Dmp-1) (quantitative polymerase chain reaction (qPCR)) were performed 72 h after irradiation. Data were analyzed by Kruskal-Wallis, ANOVA, and Tukey tests (p < 0.05). Direct light application at 4 J/cm(2) energy dose had no negative effects on cell viability, while irradiation with 2 J/cm(2) reduced cell metabolism. None of doses affected the number of viable cells compared with the control group. The two energy doses downregulated the expression of Alp; however, expression of Col-1a1 and Dmp-1 had no alteration. Cells presented change in the cytoskeleton only when irradiated with 2 J/cm(2). In conclusion, the blue LED (455 nm) irradiation, under the evaluated parameters, had no biostimulatory effects on MDPC-23 cells.

Transdentinal cell photobiomodulation using different wavelengths.

OBJECTIVE: The aim of this study was to investigate the effects of transdentinal irradiation with different light-emitting diode (LED) parameters on odontoblast-like cells (MDPC-23). METHODS AND MATERIALS: Human dentin discs (0.2 mm thick) were obtained, and cells were seeded on their pulp surfaces with complete culture medium (Dulbecco modified Eagle medium). Discs were irradiated from the occlusal surfaces with LED at different wavelengths (450, 630, and 840 nm) and energy densities (0, 4, and 25 J/cm(2)). Cell viability (methyltetrazolium assay), alkaline phosphatase activity (ALP), total protein synthesis (TP), and cell morphology (scanning electron microscopy) were evaluated. Gene expression of collagen type I (Col-I) was analyzed by quantitative polymerase chain reaction (PCR). Data were analyzed by the Mann-Whitney test with a 5% significance level. RESULTS: Higher cell viability (21.8%) occurred when the cells were irradiated with 630 nm LED at 25 J/cm(2). Concerning TP, no statistically significant difference was observed between irradiated and control groups. A significant increase in ALP activity was observed for all tested LED parameters, except for 450 nm at 4 J/cm(2). Quantitative PCR showed a higher expression of Col-I by the cells subjected to infrared LED irradiation at 4 J/cm(2). More attached cells were observed on dentin discs subjected to irradiation at 25 J/cm(2) than at 4 J/cm(2). CONCLUSION: The infrared LED irradiation at an energy density of 4 J/cm(2) and red LED at an energy density of 25 J/cm(2) were the most effective parameters for transdentinal photobiomodulation of cultured odontoblast-like cells.

Effect of radiotherapy on the eruption rate and morphology of the odontogenic region of rat incisors.

OBJECTIVE: The goal in this study was to evaluate the results of doses of 5 and 15 Gy of radiation in odontogenic region of the rats inferior mandibular-incisors by a histological analysis and the rate of eruptions. DESIGN: Animals were divided into three groups: control, radiotherapy 5 Gy and radiotherapy 15 Gy. In which tooth-eruption-rate was measured every two days. RESULTS: Animals in Group 5 Gy presented values similar to those of the control group. Animals in Group 15 Gy presented reduction in tooth-eruption-rate as of the sixth day of the experiment, vast disorganization of odontoblasts and ameloblasts, apparent reduction in cell population in the follicle region and alterations in cervical loop formation of the dental organ. CONCLUSIONS: It was concluded that there was a difference between the researched doses, and histological alteration at 15 Gy lead to statistical reduction in tooth-eruption-rate.

Effect of low-level laser therapy on odontoblast-like cells exposed to bleaching agent.

The aim of the present study was to evaluate the effect of low-level laser therapy (LLLT) on odontoblast-like MDPC-23 cells exposed to carbamide peroxide (CP 0.01%-2.21 µg/mL of H2O2). The cells were seeded in sterile 24-well plates for 72 h. Eight groups were established according to the exposure or not to the bleaching agents and the laser energy doses tested (0, 4, 10, and 15 J/cm(2)). After exposing the cells to 0.01% CP for 1 h, this bleaching solution was replaced by fresh culture medium. The cells were then irradiated (three sections) with a near-infrared diode laser (InGaAsP-780 ± 3 nm, 40 mW), with intervals of 24 h. The 0.01% CP solution caused statistically significant reductions in cell metabolism and alkaline phosphate (ALP) activity when compared with those of the groups not exposed to the bleaching agent. The LLLT did not modulate cell metabolism; however, the dose of 4 J/cm(2) increased the ALP activity. It was concluded that 0.01% CP reduces the MDPC-23 cell metabolism and ALP activity. The LLLT in the parameters tested did not influence the cell metabolism of the cultured cells; nevertheless, the laser dose of 4 J/cm(2) increases the ALP activity in groups both with and without exposure to the bleaching agent.

A novel 785-nm laser diode-based system for standardization of cell culture irradiation.

OBJECTIVE: The purpose of this study was to develop a novel device that concatenates alignment of infrared lasers and parallel procedure of irradiation. The purpose of this is to seek standardization of in vitro cell irradiation, which allows analysis and credible comparisons between outcomes of different experiments. BACKGROUND DATA: Experimental data obtained from infrared laser therapies have been strongly dependent upon the irradiation setup. Although further optical alignment is difficult to achieve, in contact irradiation it usually occurs. Moreover, these methods eventually use laser in a serial procedure, extending the time to irradiate experimental samples. METHODS: A LASERTable (LT) device was designed to provide similar infrared laser irradiation in 12 wells of a 24 well test plate. It irradiated each well by expanding the laser beam until it covers the well bottom, as occurs with unexpanded irradiation. To evaluate the effectiveness of this device, the spatial distribution of radiation was measured, and the heating of plain culture medium was monitored during the LT operation. The irradiation of LT (up to 25 J/cm(2) - 20 mW/cm(2); 1.250 sec) was assessed on odontoblast-like cells adhered to the bottom of wells containing 1 mL of plain culture medium. Cell morphology and metabolism were also evaluated. RESULTS: Irradiation with LT presented a Gaussian-like profile when the culture medium was not heated >1°C. It was also observed that the LT made it 10 times faster to perform the experiment than did serial laser irradiation. In addition, the data of this study revealed that the odontoblast-like cells exposed to low-level laser therapy (LLLT) using the LT presented higher metabolism and normal morphology. CONCLUSIONS: The experimental LASERTable assessed in this study provided parameters for standardization of infrared cell irradiation, minimizing the time spent to irradiate all samples. Therefore, this device is a helpful tool that can be effectively used to evaluate experimental LLLT protocols.

Application of high-frequency radio waves to direct pulp capping.

INTRODUCTION: In vital pulp therapy such as direct pulp capping, clinical success rates depend on achieving hemostasis in exposed pulp tissue. For hemostasis of exposed pulp tissue, gentle pressure by cotton pellets moistened with sodium hypochlorite is most commonly used. However, more rapid and reliable methods are necessary. Therefore, we focused on high-frequency radio waves (HRW). METHODS: To evaluate reparative dentin induction by HRW, we used a rat direct pulp capping model and performed hemostasis by using HRW of several strengths, covering the pulp with calcium hydroxide as a direct capping agent. After 14 or 28 days, rats were killed, and reparative dentin and pulp inflammation were investigated histologically. RESULTS: Radio wave-induced hemostasis required less time when compared with the control group. Reparative dentin with regularly arranged dentinal tubules was observed in the HRW group. CONCLUSIONS: HRW induce hemostasis and produce high-quality reparative dentin and reduced pulpal inflammation.

Low-level laser therapy influences mouse odontoblast-like cell response in vitro.

OBJECTIVE: The purpose of this study was to analyze the influence of two different irradiation times with 85 mW/cm(2) 830 nm laser on the behavior of mouse odontoblast-like cells. BACKGROUND DATA: The use of low-level laser therapy (LLLT) to stimulate pulp tissue is a reality, but few reports relate odontoblastic responses to irradiation in in vitro models. METHODS: Odontoblast-like cells (MDPC-23) were cultivated and divided into three groups: control/nonirradiated (group 1); or irradiated with 85 mW/cm(2), 830 nm laser for 10 sec (0.8 J/cm(2)) (group 2); or for 50 sec (4.2 J/cm(2)) (group 3) with a wavelength of 830 nm. After 3, 7, and 10 days, it was analyzed: growth curve and cell viability, total protein content, alkaline phosphatase (ALP) activity, calcified nodules detection and quantification, collagen immunolocalization, vascular endothelial growth factor (VEGF) expression, and real-time polymerase chain reaction (PCR) for DMP1 gene. Data were analyzed by Kruskall-Wallis test (α=0.05). RESULTS: Cell growth was smaller in group 2 (p<0.01), whereas viability was similar in all groups and at all periods. Total protein content and ALP activity increased on the 10th day with 0.8 J/cm(2) (p<0.01), as well as the detection and quantification of mineralization nodules (p<0.05), collagen, and VEGF expression (p<0.01). The expression of DMP1 increased in all groups (p<0.05) compared with control at 3 days, except for 0.8 J/cm(2) at 3 days and control at 10 days. CONCLUSIONS: LLLT influenced the behavior of odontoblast-like cells; the shorter time/smallest energy density promoted the expression of odontoblastic phenotype in a more significant way.

Effects of intrapulpal temperature change induced by visible light units on the metabolism of odontoblast-like cells.

PURPOSE: To investigate the effects of intrapulpal temperature changes induced by a quartz tungsten halogen (QTH) and a light emitting diode (LED) curing units on the metabolism of odontoblast-like cells. METHODS: Thirty-six 0.5 mm-thick dentin discs obtained from sound human teeth were randomly assigned into three groups: QTH, LED and no light (control). After placement of the dentin discs in pulp chamber devices, a thermistor was attached to the pulpal surface of each disc and the light sources were applied on the occlusal surface. After registering the temperature change, odontoblast-like cells MDPC-23 were seeded on the pulpal side of the discs and the curing lights were again applied. Cell metabolism was evaluated by the MTT assay and cell morphology was assessed by SEM. RESULTS: In groups QTH and LED the intrapulpal temperature increased by 6.4 degrees C and 3.4 degrees C, respectively. The difference between both groups was statistically significant (Mann-Whitney; P < 0.05). QTH and LED reduced the cell metabolism by 36.4% and 33.4%, respectively. Regarding the cell metabolism, no statistically significant difference was observed between both groups (Mann-Whitney; P > 0.05). However, when compared to the control, only QTH significantly reduced the cell metabolism (Mann-Whitney; P < 0.05). It was concluded that the irradiance of 0.5 mm-thick human dentin discs with a QTH in comparison to a LED curing unit promoted a higher temperature rise, which propagates through the dentin negatively affecting the metabolism of the underlying cultured pulp cells.

Trans-enamel and trans-dentinal cytotoxic effects of a 35% H2O2 bleaching gel on cultured odontoblast cell lines after consecutive applications.

AIM: To evaluate the trans-enamel and trans-dentinal cytotoxic effects of a 35% H(2)O(2) bleaching gel on an odontoblast-like cell lines (MDPC-23) after consecutive applications. METHODOLOGY: Fifteen enamel/dentine discs were obtained from bovine central incisor teeth and placed individually in artificial pulp chambers. Three groups (n = 5 discs) were formed according to the following enamel treatments: G1: 35% H(2)O(2) bleaching gel (15 min); G2: 35% H(2)O(2) bleaching gel (15 min) + halogen light (20 s); G3: control (no treatment). After repeating the treatments three consecutive times, the extracts (culture medium + gel components that had diffused through enamel/dentine discs) in contact with the dentine were collected and applied to previously cultured MDPC-23 cells (50 000 cells cm(-2)) for 24 h. Cell metabolism was evaluated by the MTT assay and data were analysed statistically (alpha = 5%; Kruskal-Wallis and Mann-Whitney U-test). Cell morphology was analysed by scanning electron microscopy. RESULTS: Cell metabolism decreased by 92.03% and 82.47% in G1 and G2 respectively. G1 and G2 differed significantly (P < 0.05) from G3. Regardless of halogen light activation, the application of the bleaching gel on the cultured odontoblast-like cells caused significantly more severe cytotoxic effects than those observed in the nontreated control group. In addition, significant morphological cell alterations were observed in G1 and G2. CONCLUSION: After three consecutive applications of a 35% H(2)O(2) bleaching agent, the diffusion of the gel components through enamel and dentine caused severe toxic effects to cultured pulp cells.

Histopathological evaluation of pulpotomy with Er,Cr:YSGG laser vs formocresol.

The purpose of this study was to histologically investigate whether pulpotomy with Er,Cr:YSGG laser is an acceptable alternative for formocresol. Pulpotomy of 48 dog's primary canine teeth was performed with formocresol or Er,Cr:YSGG laser. Histological evaluations on hematoxylin and eosin-stained pulp tissues were made by an optical microscope 7 or 60 days later. Statistical analysis was performed with Fisher's exact test, Mann-Whitney U test, and Student's t test. Seven days after pulpotomy, samples treated with laser had significantly favorable histological features in the following measures: continuity of odontoblastic layer (P<0.001), presence of hemorrhage (P<0.008), amount of inflammation (P<0.002), tissue necrosis (P<0.001), internal resorption (P<0.002), level of vascularization (P<0.002), and size of abscess (P<0.041). Similar results were observed 60 days after pulpotomy, except that the differences were not mostly significant due to natural exfoliation of 16 teeth. In conclusion, Er,Cr:YSGG laser system is an acceptable alternative for formocresol in pulpotomy of deciduous teeth.