Cytotoxicity, biocompatibility and osteoinductive profile of an MTA-hydrogel-based cement: An in vitro and animal study.

AIM: This study aimed to evaluate the cytotoxicity, biocompatibility and osteoinductive profile of a mineral trioxide aggregate (MTA)-hydrogel-based material (MTA Flow) in comparison with MTA Angelus. METHODOLOGY: Cell viability was evaluated in human periodontal ligament stem cells (hPDLSCs) using the methyl-thiazol-tetrazolium (MTT) colourimetric assay. Polyethylene tubes containing the tested materials and empty polyethylene tubes (control) were implanted in the subcutaneous tissue of Wistar rats. Cellular (lymphocyte infiltration) and extracellular events (ECM; collagen fibres) were analysed in histological sections. Immunohistochemical (collagen I, osteopontin, bone sialoprotein, bone morphogenetic protein4) analyses were also performed. RESULTS: At 24, 48 and 72 h, all tested groups showed cell viability similar to control (p > .05). Regarding biocompatibility, all groups showed similar cellular events represented by a slight inflammatory reaction characterized by hyperaemia and a mild lymphocytic inflammatory infiltrate. The analysis of lymphocytes during the time showed a decrease in these cells in the control group and a significant interaction between MTA Angelus and control (p < .001), with MTA Angelus showing a more extensive inflammatory infiltrate. Regarding fibres, an increase in content was observed in all groups during the experimental time (7, 30 and 60 days), however, no difference was detected among the experimental groups (p = .063). After 60 days, the immunoexpression of bone matrix proteins in the MTA Flow group was similar to or higher than that observed in the MTA Angelus and in the control group. CONCLUSIONS: MTA Flow showed a non-cytotoxic behaviour, biocompatibility and ability to stimulate tissue mineralization.

Antibacterial effect, cytotoxicity, and bond strength of a modified dental adhesive containing silver nanoparticles.

This study aimed to evaluate the antibacterial effect, cytotoxicity, and microtensile bond strength of an adhesive system containing silver nanoparticles (NAg). NAg was synthesized and incorporated (500 and 1000 ppm) into Scotchbond Multi-Purpose (SBMP) primer and bond. A microtensile bond test (µTBS) was performed after 24 h and 1 year. The adhesive interface was characterized using a confocal Raman microscope. The antibacterial activity was assessed using agar diffusion and biofilm inhibition assays (S. mutans). MTT assay was used to assess the cytotoxicity of NAg-conditioned culture media on human dental pulp stem cells (hDPSCs). The results were statistically analyzed using analysis of variance and Tukey's tests (α = .01). Incorporating 500 and 1000 ppm of NAg in the SBMP did not affect the µTBS after 24 h (p > 0.05). However, in the 1 year evaluation, 500 ppm presented the highest µTBS values (p < 0.05). The addition of NAg at 500 and 1000 ppm in the primer and bond led to larger inhibition halos and colony-forming units than the control (p < 0.05). For the unpolymerized and polymerized groups, the combination of primer and bond presented the highest cytotoxic effects on hDPSCs (p < 0.05). In conclusion, incorporating 500 or 1000 ppm of NAg into an etch-and-rinse adhesive system led to an antibacterial effect without altering the cytotoxicity. SBMP at 500 ppm presented a higher µTBS at 1 year.

Photobiomodulation treatments drive osteogenic versus adipocytic fate of bone marrow mesenchymal stem cells reversing the effects of hyperglycemia in diabetes.

Diabetes mellitus (DM) is a chronic metabolic disease that affects bone metabolism, which can be related to a reduced osteogenic potential of bone marrow mesenchymal stem cells (BM-MSCs). MSCs from diabetic rats (dBM-MSC) have shown a tendency to differentiate towards adipocytes (AD) instead of osteoblasts (OB). Since photobiomodulation (PBM) therapy is a non-invasive treatment capable of recovering the osteogenic potential of dBM-MSCs, we aimed to evaluate whether PBM can modulate MSC's differentiation under hyperglycemic conditions. BM-MSCs of healthy and diabetic rats were isolated and differentiated into osteoblasts (OB and dOB) and adipocytes (AD and dAD). dOB and dAD were treated with PBM every 3 days (660 nm; 5 J/cm2; 0.14 J; 20 mW; 0.714 W/cm2) for 17 days. Cell morphology and viability were evaluated, and cell differentiation was confirmed by gene expression (RT-PCR) of bone (Runx2, Alp, and Opn) and adipocyte markers (Pparγ, C/Ebpα, and C/Ebpß), production of extracellular mineralized matrix (Alizarin Red), and lipid accumulation (Oil Red). Despite no differences on cell morphology, the effect of DM on cells was confirmed by a decreased gene expression of bone markers and matrix production of dOB, and an increased expression of adipocyte and lipid accumulation of dAD, compared to heatlhy cells. On the other hand, PBM reversed the effects of dOB and dAD. The negative effect of DM on cells was confirmed, and PBM improved OB differentiation while decreasing AD differentiation, driving the fate of dBM-MSCs. These results may contribute to optimizing bone regeneration in diabetic patients.

Efficacy of photobiomodulation therapy on healing of ionizing irradiated bone: a systematic review of in vivo animal studies.

This systematic review aimed to answer the research focused question: What are the effects of photobiomodulation (PBM) therapy on bone healing after ionizing irradiation in animal models? The EMBASE, LILACS, LIVIVO, PubMed, Scopus, and Web of Science databases, including gray literature, were searched using the following keywords: "Head and Neck Neoplasms"; "Ionizing Radiation"; "Low-Level Light Therapy"; and "Bone regeneration", focusing on the primary studies that assessed the effects of PBM therapy on animal models of irradiated bone. Six studies have met the eligibility criteria and presented an overall regular quality according to the risk of bias assessment tools. All the studies utilized rat animal model and near-infrared laser PBM at low power output setting. Most of the studies showed increased new bone formation, osteocytes, osteoblasts, and vascularization networking, as a result of PBM therapy. However, only one out of the six studies has not shown any differences in bone healing in both lased and non-lased animal groups. Nevertheless, PBM therapy is a potential tool to improve bone healing induced by ionizing radiation. However, due to the scarce number of studies and the great variability of laser parameters and treatment protocols, a clear conclusion cannot be drawn. Hence, extensive preclinical in vivo studies are warranted to ensure these beneficial effects have been addressed prior to translational clinical trials.

Photobiomodulation as an adjunctive therapy for alveolar socket preservation: a preliminary study in humans.

Bone remodeling results in loss of alveolar bone height and thickness. Photobiomodulation (PBM) based on photochemical stimulation by low-intensity lasers emerges as an adjunctive therapy for alveolar socket preservation. Our study aimed to evaluate the effects of PBM therapy on alveolar bone repair. Twenty healthy patients in need of bilateral extraction of lower molars were enrolled in this split-mouth randomized and blind clinical trial. The extraction sites were randomly selected to receive either the PBM therapy with a CW GaAIAs diode laser (808 nm; 0.028 mm2; 0.1 W; 3.6 W/cm2; 89 J/cm2; 2.5 J/point) or no treatment (Control). Bone biopsies were harvested 45 days after the dental extraction and evaluated using micro-computerized tomography (µCT), morphometric, and histological analysis. Data were compared using the paired t test, and the level of significance was set at 5%. Bone surface (p = 0.029), bone surface/total volume (p = 0.028), trabecular number (p = 0.025), and connectivity density (p = 0.029) were higher at the PBM group compared with Control. The histological observations confirmed the µCT findings. PBM samples exhibited higher number of organized and connected bone trabeculae along with higher density of blood vessels than Control. Control samples displayed a dense and highly cellular connective tissue at the central area accompanied by the presence of immature bone trabeculae at the periphery. Our results indicated that the PBM therapy improved the newly bone trabeculae formation and their connectivity which increased bone surface, indicating the positive effect of the laser on alveolar human socket repair.

Aquaporin 1, 3, and 5 Patterns in Salivary Gland Mucoepidermoid Carcinoma: Expression in Surgical Specimens and an In Vitro Pilot Study.

Salivary gland aquaporins (AQPs) are essential for the control of saliva production and maintenance of glandular structure. However, little is known of their role in salivary gland neoplasia. Salivary gland tumors comprise a heterogeneous group of lesions, featuring variable histological characteristics and diverse clinical behaviors. Mucoepidermoid carcinoma (MEC) is the most common salivary gland malignancy. The aim of this study was to evaluate the expression of AQP1, AQP3, and AQP5 in 24 MEC samples by immunohistochemistry. AQP1 expression was observed in vascular endothelium throughout the tumor stroma. AQP3 was expressed in epidermoid and mucosal cells and AQP5 was expressed in mucosal cells of MEC. These proteins were expressed in the human MEC cell line UH-HMC-3A. Cellular ultrastructural aspects were analyzed by electron microscopy to certificate the tumor cell phenotype. In summary, our results show that, despite the fact that these molecules are important for salivary gland physiology, they may not play a distinct role in tumorigenesis in MEC. Additionally, the in vitro model may offer new possibilities to further investigate mechanisms of these molecules in tumor biology and their real significance in prognosis and possible target therapies.

Examining tumor modulating effects of photobiomodulation therapy on head and neck squamous cell carcinomas.

Photobiomodulation (PBM) therapy is an effective method for preventing and managing oral mucositis (OM) in head and neck squamous cell carcinoma (HNSCC) patients undergoing radiotherapy alone or in combination with chemotherapy. However, the potential effects of PBM therapy on premalignant and malignant cells eventually present in the treatment site are yet unknown. The aim of this systematic review was to analyze the effects of PBM therapy on HNSCC. A literature search was conducted in four indexed databases as follows: MEDLINE/PubMed, EMBASE, Web of Science, and Scopus. The databases were reviewed for papers published up to and including in October 2018. In vitro and in vivo studies that investigated the effects of PBM therapy on HNSCC were selected. From the 852 initially gathered studies, 15 met the inclusion criteria (13 in vitro and 2 in vivo). Only three in vitro studies were noted to have a low risk of bias. The included data demonstrated wide variations of study designs, PBM therapy protocols, and study outcomes. Cell proliferation and viability were the primary evaluation outcome in the in vitro studies. Of the 13 in vitro studies, seven noted a positive effect of PBM therapy on inhibiting or preventing an effect on HNSCC tumor cells, while six studies saw increased proliferation. One in vivo study reported increased oral SCC (OSCC) progression, while the other observed reduced tumor progression. Overall, the data from the studies included in the present systematic review do not support a clear conclusion about the effects of PBM therapy on HNSCC cells.

Short-term evaluation of photobiomodulation therapy on the proliferation and undifferentiated status of dental pulp stem cells.

The aim of this in vitro study was to analyze the effect of photobiomodulation therapy (PBMT) on the proliferation and undifferentiating status of stem cell from human exfoliated deciduous teeth (SHEDs). PBMT was carried out with an aluminum gallium indium phosphide (InGaAlP) diode laser in contact and punctual mode (continuous wave, 660 nm, 20 mW, 0.028 cm2, and average energy densities of 1 (1 s), 3 (4 s), 5 (7 s), 10 (14 s), 15 (21 s), or 20 (28 s) J/cm2 per point). The immunoprofile of the SHEDs was analyzed using flow cytometry. Cell proliferation was assessed by the MTT reduction assay. Gene expressions of mesenchymal stem cell markers (OCT4, Nestin, CD90, and CD105) were assessed by RT-qPCR 48 h after PBMT. Data were compared by analysis of variance (ANOVA) and Tukey's test (p ≤ 0.05). Cells cultured under nutritional deficit and treated with PBMT at 5 J/cm2 presented similar cell growth than those of positive control group. Cell growth was significantly higher than those of other groups. Mesenchymal stem cell gene markers were still expressed after PBMT at 5 J/cm2. In a short-term analysis, PBMT increases the number of stem cells with no interference in the undifferentiated state of the irradiated cells, which opens wide possibilities for application in tissue regeneration.

Cell sheets of human dental pulp stem cells for future application in bone replacement.

OBJECTIVES: To analyze the potential of human dental pulp stem cells (hDPSCs) for maintaining their undifferentiated status and osteogenic differentiation capacity when arranged in cell sheets (CSs) for future application in bone replacement. MATERIALS AND METHODS: CSs were formed after being induced for 10-15 days by clonogenic medium containing additional vitamin C (20 µg/ml). The cell viability of hDPSC4s in the CSs was followed until 96 h using the Live/Dead® assay. The cells of the CSs were enzymatically dissociated and then compared with the original hDPSC4s. The two cell types were characterized immunophenotypically by flow cytometry using specific mesenchymal stem cell-associated markers (CD105, CD146, CD44, STRO-1, and OCT3/4) and non-associated markers (CD34, CD45, and CD14). Osteogenic differentiation was analyzed with the Alizarin red assay. RESULTS: Living cells were observed until 96 h in the CSs. Both cell types exhibited osteogenic differentiation and expressed the specific undifferentiated MSC-associated markers. Cells spontaneously detached from the CSs attached and proliferated at the bottom of the culture dishes. CONCLUSIONS: Cells in the hDPSC4s cell sheets survived for at least 96 h. Moreover, the cells in the cell sheets retained their stemness and their osteogenic differentiation potential. CLINICAL RELEVANCE: Cell sheets of hDPSCs could be employed as natural tri-dimensional structures for treating bone loss. This technique would be useful particularly for critical bone defects or any type of bone defects in patients carrying diseases that impair bone regeneration, such as diabetes mellitus, medication-related osteonecrosis of the jaw (MRONJ), and osteoporosis.

Relationship between toothpastes properties and patient-reported discomfort: crossover study.

OBJECTIVES: This study aims to correlate patient-reported reactions with in vitro analyses of the pH, abrasive quality, and cytotoxicity of four toothpastes. MATERIALS AND METHODS: One hundred twenty-one patients received non-identified samples of toothpaste to be used for 6 days and answered a questionnaire about their sensations. In vitro analysis: the pH of toothpastes was measured with a pH meter. The abrasivity of toothpastes was evaluated against composite resin specimens (n = 10). A toothbrushing machine was used to simulate wear, which was indirectly measured by mass loss using a scale. Cell culture media conditioned with toothpaste were used to assess the cytotoxicity. Confluent cells were kept in contact with the conditioned media or control for 24 h. The cell viability was measured using the 3-(bromide, 4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium (MTT)-reduction assay. The obtained data on the pH, weight loss, and cell viability were compared by ANOVA/Tukey's tests (p < 0.05). RESULTS: With the exception of the bleaching effect paste, the Oral B® paste produced the highest frequencies of irritation reports, tooth sensitivity, taste discomfort, and texture discomfort in the clinical study; patients also reported rougher teeth, soft tissue peeling, dry mouth, thrush, tingling, and taste changes in response to this paste. The in vitro analysis demonstrated that Oral B® had the lowest pH, the highest abrasivity, and produced the lowest cell viability (p < 0.01). CONCLUSION: Results suggest that low pH toothpastes that are highly abrasive and cytotoxic may cause undesirable reactions in patients. CLINICAL RELEVANCE: Toothpaste's properties should be well known for indication to patient therefore minimizing discomfort reports.

Evaluation of pain, jaw movements, and psychosocial factors in elderly individuals with temporomandibular disorder under laser phototherapy.

Few studies have been carried out on the application of laser phototherapy (LPT) for treating painful temporomandibular disorder (TMD) in elderly population that is growing worldwide. The aim of the present study was to evaluate the pain, jaw movements, and psychosocial factors in ten elderly patients with painful TMD before and after LPT. All patients were evaluated before and after LPT by using the Research Diagnostic Criteria for temporomandibular disorders (RDC/TMD) axes I and II. For pain assessment, a visual analogue scale (VAS) was used. The LPT was carried out with an GaAlAs diode laser (780 nm; spot size 0.04 cm(2)) in punctual and contact mode. Two settings of irradiations were applied as follows: in patients presenting myofascial pain, 10 mW, 5 J/cm(2), 20 s, 0.2 J per application point; and in patients with joint TMD, 70 mW, 105 J/cm(2), 60 s on five points, 4.2 J per point. Two sessions of LPT were carried out per week over four consecutive weeks, in the total of eight sessions. Data was statistically analyzed (p < 0.05). Significant pain reduction was found in all patients. There were increase in maximum mouth opening without pain and reduction in muscle pain during right and left lateral excursion. A significant reduction in chronic pain severity (p = 0.02) and significant improvements in depression (p = 0.038) and nonspecific physical symptoms with pain (p = 0.0167) were observed. The present findings indicate that LPT is able to promote pain relief and improvement of jaw movements in elderly patients with TMD, with a positive effect on psychosocial aspects.

Influence of CO2 (10.6 µm) and Nd:YAG laser irradiation on the prevention of enamel caries around orthodontic brackets.

One possible undesirable consequence of orthodontic therapy is the development of incipient caries lesions of enamel around brackets. The aim of this study was to compare the effects of CO2 (λ = 10.6 µm) and Nd:YAG (λ = 1,064 nm) lasers associated or not with topical fluoride application on the prevention of caries lesions around brackets. Brackets were bonded to the enamel of 65 premolars. The experimental groups (n = 13) were: G1--application of 1.23% acidulated fluoride phosphate gel (AFP, control); G2--Nd:YAG laser irradiation (0.6 W, 84.9 J/cm(2), 10 Hz, 110 µs, contact mode); G3--Nd:YAG laser irradiation associated with AFP; G4--CO2 laser irradiation (0.5 W, 28.6 J/cm(2), 50 Hz, 5 µs, and 10 mm focal distance); and G5--CO2 laser irradiation associated with AFP. Quantitative light-induced fluorescence was used to assess enamel demineralization. The data were statistically compared (α = 5%). The highest demineralization occurred in the Nd:YAG laser group (G2, 26.15% ± 1.94). The demineralization of all other groups was similar to that of the control group. In conclusion, CO2 laser alone was able to control enamel demineralization around brackets at the same level as that obtained with topical fluoride application.

Laser phototherapy enhances mesenchymal stem cells survival in response to the dental adhesives.

BACKGROUND: We investigated the influence of laser phototherapy (LPT) on the survival of human mesenchymal stem cells (MSCs) submitted to substances leached from dental adhesives. METHOD: MSCs were isolated and characterized. Oral mucosa fibroblasts and osteoblast-like cells were used as comparative controls. Cultured medium conditioned with two adhesive systems was applied to the cultures. Cell monolayers were exposed or not to LPT. Laser irradiations were performed using a red laser (GaAlAs, 780 nm, 0.04 cm(2), 40 mW, 1 W/cm(2), 0.4 J, 10 seconds, 1 point, 10 J/cm(2)). After 24 h, cell viability was assessed by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide reduction assay. Data were statistically compared by ANOVA followed by Tukey's test (P < 0.05). RESULTS: Different cell types showed different viabilities in response to the same materials. Substances leached from adhesives were less cytotoxic to MSCs than to other cell types. Substances leached from Clearfil SE Bond were highly cytotoxic to all cell types tested, except to the MSCs when applied polymerized and in association with LPT. LPT was unable to significantly increase the cell viability of fibroblasts and osteoblast-like cells submitted to the dental adhesives. CONCLUSION: LPT enhances mesenchymal stem cells survival in response to substances leached from dental adhesives.

Role of rhBMP-2 and rhBMP-7 in the metabolism and differentiation of osteoblast-like cells cultured on chemically modified titanium surfaces.

This study analyzed the role of recombinant human bone morphogenetic protein 2 (rhBMP-2) and recombinant human bone morphogenetic protein 7 (rhBMP-7) in the adhesion and differentiation of rat osteoblast-like (osteo-1) cells cultured on chemically modified titanium surfaces. Osteo-1 cells were cultured on chemically modified (modified sandblasted and acid-etched) titanium surfaces in 3 different types of medium: control, medium supplemented with 20 ng/mL rhBMP-2, and medium supplemented with 20 ng/mL rhBMP-7. The following parameters were evaluated: cell adhesion after 24 hours; total protein content; collagen content and alkaline phosphatase (AP) activity after 7, 14, and 21 days; and calcified nodule formation after 21 days. The addition of rhBMP-2 or rhBMP-7 did not influence cell adhesion (P = .1175). Cell differentiation was influenced by rhBMP-2, as demonstrated by a significant increase in collagen content after 7 days of culture (P < .0001) and a significant decrease in AP activity after 21 days (P < .0001). The addition of rhBMP-7 only influenced AP activity, and a significant increase was observed after 21 days (P < .0001). Within the limitations of the study, we conclude that the presence of rhBMP-2 or rhBMP-7 did not influence cell adhesion to chemically modified titanium surfaces but provided an additional stimulus during the differentiation of rat osteo-1 cells cultured on this type of surface.

New uses for rehabilitation protocol for oral sinus communications in ARONJ patients.

This report presents a new use for rehabilitation protocol for oral sinus communications in patients with antiresorptive agent-induced osteonecrosis of the jaw. The treatment plan consisted of constructing an atraumatic complete denture with rounded edges, made with nontoxic resin, to prevent any injury to the mucosa and recurrence of the disease. The patient was followed up for 4 years, without any complications, and was socially reintegrated by resuming the normal life he experienced before tooth loss.

Osteoradionecrosis in a Torus Mandibularis Treated by Antimicrobial Photodynamic Therapy: A Case Report.

Background: Osteoradionecrosis (ORN) of the jaws is a late complication after radiotherapy to head and neck cancer. Objective: To describe a rare case of ORN of the torus mandibularis that was successfully managed exclusively with antimicrobial photodynamic therapy (aPDT). Case report: A 72-year-old man presented an exposed necrotic bone observed in the torus mandibularis, extending to the lingual alveolar ridge with no edema nor suppuration. The treatment provided a noninvasive treatment leading to spontaneous sequestrectomy of the torus in 2 weeks with complete mucosal repair in 5 weeks and absence of lesion signs and/or symptoms even after 6 months of follow-up. Conclusions: The aPDT indicated to be a satisfactory treatment for ORN affecting torus mandibularis, a region with surgical limitations, avoiding surgery.

Tooth bleaching induces changes in the vascular permeability of rat incisor pulps.

PURPOSE: To evaluate the inflammatory response in dental pulps of rat incisors subjected to tooth bleaching protocols with different HP concentrations and application times. METHODS: 42 incisors from Wistar rats were submitted to tooth bleaching using concentrations of 25% or 35% HP for treatment times of 15, 30 or 45 minutes. Four non-bleached teeth were used as controls. The animals received an intravenous injection of India ink immediately after the bleaching procedure and were sacrificed 1 hour later. Six bleached teeth from each group and three controls were made transparent, and one sample from each group was processed for histological analysis. The data were statistically analyzed using Kruskal Wallis and Dunn's tests (P < 0.05). RESULTS: The amount of dental pulp ink content was significantly higher in the samples that were bleached with 35% HP for 30 minutes and with both HP concentrations (25 and 35%) for 45 minutes than in the controls. For the samples bleached with the same HP concentration, the ink content was higher in samples that were bleached for 45 minutes. These results indicate that HP tooth bleaching can induce an increase in vascular permeability in rat incisors. Importantly, this increase is more dependent on the length of the bleaching procedure than on the concentration of the bleaching agent.

Toward personalized cell therapies by using stem cells: seven relevant topics for safety and success in stem cell therapy.

Stem cells, both embryonic and adult, due to the potential for application in tissue regeneration have been the target of interest to the world scientific community. In fact, stem cells can be considered revolutionary in the field of medicine, especially in the treatment of a wide range of human diseases. However, caution is needed in the clinical application of such cells and this is an issue that demands more studies. This paper will discuss some controversial issues of importance for achieving cell therapy safety and success. Particularly, the following aspects of stem cell biology will be presented: methods for stem cells culture, teratogenic or tumorigenic potential, cellular dose, proliferation, senescence, karyotyping, and immunosuppressive activity.

Bactericidal effects of two parameters of Er:YAG laser intracanal irradiation: ex-vivo study.

The success of endodontic treatment depends on the complete elimination of microorganisms from the root canal system, thus the search for new procedures to eliminate them is justified. The aim of this study was to assess bacterial reduction after intracanal irradiation with the Er:YAG laser. The canals of 70 extracted human maxillary canines were prepared up to file #40 using 1% NaOCl, irrigated with 17% EDTA, and then washed with physiological solution activated by ultrasound. The roots were sterilized by autoclaving, inoculated with 10 µl of a suspension containing 1.5 × 10(8) CFU/ml of Enterococcus faecalis ATCC 29212 and incubated at 37°C for 72 h. The canals were irradiated with the Er:YAG laser using two energy settings: 60 mJ and 15 Hz, and 100 mJ and 10 Hz. The remaining bacteria were counted immediately and 48 h after laser irradiation. The results showed a high bacterial reduction at both time points. With 60 mJ and 15 Hz there was an immediate reduction of 99.73% and the reduction was 77.02% after 48 h, and with 100 mJ and 10 Hz there was an immediate reduction of 99.95% and the reduction was 84.52% after 48 h. Although the best results were observed with 100 mJ of energy, the difference between the two settings was not statistically significant. The count performed 48 h after irradiation showed that E. faecalis were able to survive, and can grow even from small numbers.

Cytotoxicity evaluation of Curcuma zedoaria (Christm.) Roscoe fluid extract used in oral hygiene products.

OBJECTIVE: This in vitro study evaluated the cytotoxic effects of the Curcuma zedoaria (Christm.) Roscoe (popular name: zedoary) fluid extract, as used in preparations for oral hygiene, mostly for anti-septic purposes. MATERIALS AND METHODS: The cell viability and cell growth were assessed by Trypan blue dye exclusion assay using the LMF cell line derived from oral mucosa. Cell viability (short-term assay) was measured 0, 6, 12 and 24 h after contact with the fluid extract. Cell growth (long-term assay) was analyzed in 1, 3, 5 and 7 days. The experimental groups were those testing the fluid extract obtained from the zedoary rhizome and the extractor liquid (ethanol 70° GL) in the concentrations of 0.01-0.0001% v/v. Fresh DMEM were used in the control cultures. RESULTS: Short-term assay-all studied cultures maintained stable cell viability; Long-term assay-there was progressive cell growth in all studied cultures. CONCLUSION: According to the results, the zedoary fluid extract presents low cytotoxicity and probably can be used in the oral hygiene products.