The direct cytotoxic effects of medicaments used in endodontic regeneration on human dental pulp cells.

AIM: The purpose of this in vitro study was to evaluate the effects of intracanal medicaments commonly used in endodontic regeneration on the survival of human dental pulp cells (DPCs). METHODS: DPCs were cultured and exposed to either no medicament treatment or low concentrations (0.3-5 mg ml(-1) ) of calcium hydroxide [Ca(OH)2 ], triple antibiotic paste (TAP), or double antibiotic paste (DAP) for 3 days. After that, toxicity to the DPCs was determined by lactate dehydrogenase activity assays (LDH) and cell proliferation was measured by colorimetric assays (WST-1). Two-way anova followed by Fisher's protected least significant differences was used for statistical analyses (α = 0.05). RESULTS: The group-by-concentration interactions were significant for the LDH and WST-1 assays (P < 0.0001). For the LDH assays, only the highest tested concentration (5 mg ml(-1) ) of Ca(OH)2 and TAP caused significant toxicity to the DPCs compared with the untreated control, while four tested concentrations of DAP (0.5, 1, 2.5, and 5 mg ml(-1) ) caused significant toxicity to the DPCs compared with the untreated control. For the WST-1 assays, the highest concentrations that did not negatively affect the proliferation rate of DAP, TAP and Ca(OH)2 were 0.3, 2, and 2.5 mg ml(-1) , respectively. CONCLUSION: The low concentrations of intracanal medicaments tested in this study were not cytotoxic in cultured cells. However, these concentrations are much lower than the concentrations that have been advocated in endodontic regeneration. Furthermore, the negative effects of TAP on DPCs were detected at lower concentrations by using the WST-1 assays than by measuring the LDH release.

Cytotoxicity of QMix™ endodontic irrigating solution on human bone marrow mesenchymal stem cells.

BACKGROUND: Debridement and disinfection of the root canal system is a crucial step in endodontic procedures. The effectiveness of irrigation relies on both the mechanical flushing action and the ability of irrigants to dissolve tissue and kill bacteria. The objective of the present study is to evaluate and compare the cytotoxicity of QMix™ root canal irrigating solution on immortalized human bone marrow mesenchymal stem cells (hTERT-MSC-C1) and to compare it with that of sodium hypochlorite (NaOCl). METHODS: Immortalized human bone marrow mesenchymal stem cells (hTERT-MSCs) were exposed to QMix™ and NaOCl. Cell viability was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and alamarBlue assays. The cell morphology was studied after two hours of exposure to QMix™ and NaOCl. Scanning electron microscopy (SEM) analyses were performed after 2- and 4-hour incubation periods. Finally, ethidium bromide/acridine orange (EB/AO) fluorescent stain was applied to the cells in the 8-chamber slides after they were incubated with the testing agents for 2 hours to detect live and dead cells. The observations were tabulated and analyzed statistically. RESULTS: QMix™ exposure resulted in a significantly higher percentage of cell viability than NaOCl in the MTT and alamarBlue assays at three time points compared to the control. The SEM analysis demonstrated minimal morphological changes associated with cells that were exposed to the QMix™ solution, with little shrinkage and fragmentation of the cell wall. The live/dead analysis showed that the number of live cells after exposure to QMix™ was similar to that of the untreated control. No cell structure could be observed with the NaOCl group, indicating cell lysis. CONCLUSION: Both the QMix™ and NaOCl solutions were toxic to human bone marrow MSCs. Each solution might have induced cell death in a different way as evidenced in the cell viability, SEM and fluorescent studies. The slower cell death induced by QMix™ might therefore be less aggressive and more acceptable to living tissues.

An in vitro evaluation of the cytotoxicity of varying concentrations of sodium hypochlorite on human mesenchymal stem cells.

AIM: To evaluate and compare the cytotoxicity of various concentrations of sodium hypochlorite on immortalized human bone marrow mesenchymal stem cells (MSCs). MATERIALS AND METHODS: The 5.25 percent sodium hypochlo-rite (NaOCl) at concentrations of 0.5, 0.1, 0.025, 0.0125, and 0.005 mg/ml were used to assess the cytotoxic effect on MSCs. Immortalized human bone marrow mesenchymal stem cells (hTERT-MSCs) were exposed to NaOCl at 5 different concentrations. Cell viability was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and alamarBlue assays. The cell morphology changes were assessed with scanning electron microscopy (SEM) after exposure to 2, 4, and 24 hour incubation. The ethidium bromide/acridine orange (EB/ AO) fuorescent stain was applied to the cells in the 8-chamber slides after they were incubated with the testing agents for 2 and 4 hours to detect live and dead cells. The observations were quantitatively and qualitatively analyzed. RESULTS: The cell viability study using MTT assay and AB assay showed significant reduction with varying concentration at 2 and 4 hours incubation period. The cell viability decreased with the higher percentage of NaOCl. The exposure time also revealed an inverse relation to the cell viability. The SEM analysis showed reduction in the number of cells and morphological alterations with 0.5 mg/ml at 2 and 4 hours compared to 0.025 mg/ml NaOCl. Destruction of the cells with structural alterations and lysis was evident under fuorescence microscope when the cells were exposed to 0.5 mg/ml NaOCl. CONCLUSION: Within the limitations of this in vitro study it can be concluded that NaOCl is toxic to the human bone marrow MSCs. The cell lysis was evident with higher concentration of sodium hypochlorite. From the observations, it can be concluded that a lower concentration of NaOCl may be used as endodontic irrigant due to its cytotoxic properties. Further studies are mandatory to evolve a consensus on the optimal concentration of sodium hypochlorite to be used as endodontic irrigant.

Antifungal and genotoxic effects of Thymus serpyllum as a root canal irrigant.

OBJECTIVES: The aim of this study was the assessment of the efficiency of the ethyl acetate (EthOAc) extract of Thymus serpyllum against Candida albicans and to compare it with sodium hypochlorite (NaOCl) and chlorhexidine (CHX), as well as their genotoxic effect. MATERIAL AND METHODS: The antifungal effectiveness of the EthOAc extract of Thymus serpyllum was determined using the agar disk diffusion method. The inhibition zones induced by the EthOAc extract were compared after 5 min, 60 min, and 24 h to those induced by standard solutions (2% CHX and 2% NaOCl). An in vitro genotoxicity assay was performed in cultured lymphocytes from the blood of human volunteers to observe micronuclei formation. Statistical analysis of the results was performed using the Kruskal-Wallis test and one-way analysis of variance. RESULTS: The inhibition zone of combination of CHX with EthOAc extract of Thymus serpyllum against C. albicans was 29.7 mm after 5 min, 28.3 mm after 60 min, and 29 mm after 24 h. The inhibition zone of NaOCl in combination with EthOAc extract of Thymus serpyllum against C. albicans was 0 mm. The EthOAc extract of Thymus serpyllum did not show a genotoxic effect on lymphocyte cells. CONCLUSIONS: The EthOAc extract of Thymus serpyllum in combination with CHX may be a useful root canal disinfection in endodontic therapy.

Sensitivity of human dental pulp cells to eighteen chemical agents used for endodontic treatments in dentistry.

To determine the adverse effects against human dental pulp tissue, the sensitivity of human dental pulp cells (D824 cells) to 18 chemical agents used for endodontic treatments in dentistry was examined. The cytotoxicity, as determined by a decrease in colony-forming ability of cells treated with the chemical agents, increased as the concentration increased. As a quantitative measure of the cytotoxic effect, LC(50), the concentration which induces a 50% lethality, was extrapolated from the concentration-response curves. The rank of the chemical agents according to their cytotoxic effect (LC(50)) was sodium arsenite > formaldehyde > hydrogen peroxide > zinc oxide > thymol ≈ iodoform ≈ eugenol > guaiacol > ethylenediaminetetraacetic acid ≈ iodine > procaine > lidocaine ≈ chloramphenicol ≈ m-cresol > calcium hydroxide ≈ sodium hypochlorite ≈ phenol ≈ p-phenolsulfonic acid. To compare the cytotoxicity and the levels of apoptosis and mRNA expression of five genes related to the function of dental pulp tissue, D824 cells treated with the LC(50) concentrations of chemical agents were assayed by the TUNEL method and quantitative reverse transcription polymerase chain reaction analysis, respectively. The inducibility of apoptotic cells and the level of mRNA expression of the genes varied with the chemical agents, indicating that both effects occurred independent of the rank of cytotoxic effect of the chemical agents. The results not only provide information concerning cytotoxicity of various chemical agents to human dental pulp cells, but also show an insight into the diversity of the pharmacodynamic action of the chemical agents.

Propolis and commonly used intracanal irrigants. Comparative evaluation of inflammatory potential.

AIM: The present study evaluated the inflammatory/irritant potential of propolis in comparison with commonly used intracanal irrigants such as chlorhexidine and calcium hydroxide, with normal saline solution as control using an animal (Wistar rats) model. METHOD: 2% Evans blue was intravenously injected into the lateral caudal vein. 0.1 ml each of the test solutions was intradermally injected into the experimental sites designed on their shaved backs. The animals were then sacrificed after 1 1/2 and 3 hours respectively. Each piece of skin containing the injected solution was excised, immersed in 4 ml formamide and incubated at 45 degrees C for 72 hours. After filtration with glass wool, optical density(OD) was measured using a spectro-photometer and analyzed statistically. RESULTS: At 620 nm irrespective of time, the mean optical density with Calcium Hydroxide was found to be maximum (0.197 +/- 0.095) while that with DMSO Propolis was found to be minimum (0.070 +/- 0.016). Both at 90 min and 180 min, the mean optical density with Calcium Hydroxide was found to be maximum. CONCLUSIONS: On short-term evaluation, maximum inflammation was seen with calcium hydroxide followed by chlorhexidine and DMSO extract of propolis. Minimum inflammation was seen with sterile physiologic saline. With progress of time, maximum inflammation was seen with calcium hydroxide followed by chlorhexidine and DMSO extract of propolis which was non-significant.

The cytotoxic effects of various endodontic irrigants on the viability of dental mesenchymal stem cells.

This study aimed to evaluate cytotoxic effects of various irrigation solutions used in regenerative endodontic treatments (RETs) on mesenchymal stem cells, and further examine the long-term effect of hypochlorous acid (HOCl) on the cell viability and alkaline phosphatase (ALP) activity. Stem cells were exposed to various concentrations of NaOCl, EDTA, chlorhexidine (CHX), etidronic acid (HEDP)/NaOCl combination and HOCl. HOCl was tested for its effects on ALP activity up to 21 days. Additionally, cell viability was measured fluorescently using calcein AM. The most cytotoxic irrigant was CHX even with the lowest concentration. NaOCl and HEDP/NaOCl with 1:100 dilution decreased viability to around 40%. HOCl showed the lowest cytotoxicity among all tested irrigants. HOCl also showed no significant reduction in ALP activity compared with the controls. The cytotoxicity of endodontic irrigants was time and concentration dependent. HOCl demonstrated promising results regarding viability and ALP activity, since RETs require host stem cell survival.

A laboratory evaluation of the antibacterial and cytotoxic effect of Liquorice when used as root canal medicament.

AIM: To evaluate the antibacterial and cytotoxic effects of Liquorice as a root canal medicament and to compare its action to the commonly used root canal medicament calcium hydroxide Ca(OH)(2). METHODOLOGY: The antibacterial effect of Liquorice and Ca(OH)(2) either separately or in combination was investigated against Enterococcus faecalis. Agar-well diffusion methods, broth microdilution tests and biofilm susceptibility assays were used to determine the antibacterial activity. Human periodontal ligament fibroblast tissue culture was used to assess the cytotoxicity of the preparations under investigation. RESULTS: Liquorice extract either by itself or in combination with Ca(OH)(2) had a significant inhibitory effect against Enterococcus faecalis compared with that of Ca(OH)(2) alone. The use of Liquorice extract followed by Liquorice/Ca(OH)(2) mixture retained significantly more viable periodontal ligament cells than Ca(OH)(2) , which had a strong lethal effect on the cells. CONCLUSION: Liquorice extract either separately or as Liquorice/Ca(OH)(2) mixture had a potent bactericidal effect against Enterococcus faecalis and retained compatibility with fibroblasts in tissue culture compared to the commonly used root canal medicament Ca(OH)(2).

Cytotoxicity evaluation of calcium hypochlorite and other commonly used root canal irrigants against human gingival fibroblast cells: An in vitro evaluation.

BACKGROUND: The conventional endodontic therapy primarily focuses on biomechanical preparation, which is achieved by the application of various intracanal irrigants and intracanal medicaments. One of the most commonly used intracanal irrigants - sodium hypochlorite (NaOCl) - has already been proven to have an antimicrobial effect as well as the ability to dissolve tissues in the areas where files cannot reach. One of the recently used irrigants having a promising effect is calcium hypochlorite (Ca(OCl)2), which has been shown to be relatively more stable than NaOCl and has much more chlorine ions. OBJECTIVES: The aim of this study was to assess the individual cytotoxicity of various root canal irrigants and the combined cytotoxicity of NaOCl and Ca(OCl)2 with ethylenediaminetetraacetic acid (EDTA) against human gingival fibroblast (hGF) cells. MATERIAL AND METHODS: The evaluation of the individual cytotoxicity was carried out with regard to the following root canal irrigants: NaOCl; Ca(OCl)2; and chlorhexidine (CHX). The evaluation of the combined cytotoxicity regarded NaOCl/EDTA and Ca(OCl)2/EDTA. The concentrations used were 0.025%, 0.050%, 0.10%, and 0.20%. The cytotoxicity against hGF cells was examined within a timeframe of 6 h and 24 h with the use of the sulforhodamine B (SRB) assay. RESULTS: It was observed that Ca(OCl)2 had a mean absorbance rate of 0.315 ±0.02, 0.294 ±0.03, 0.265 ±0.03, and 0.240 ±0.02 at 0.025%, 0.050%, 0.10%, and 0.20%, respectively. In combination with EDTA, the mean absorbance rate was 70.12 ±2.9, 67.42 ±4.3, 64.35 ±3.6, and 61.58 ±4.1 at 0.025%, 0.050%, 0.10%, and 0.20%, respectively. The cytotoxic effect of the root canal irrigants on hGF cells was observed to be statistically significant (p < 0.05). CONCLUSIONS: Calcium hypochlorite is less cytotoxic than NaOCl, and when used in combination with EDTA, it was shown to have its cytotoxic effect on hGF cells reduced to a great extent.

Chlorhexidine as a canal irrigant: a review.

The use of an irrigant during root canal therapy is an important factor in the cleaning and disinfecting of the root canal system. While sodium hypochlorite has been used for decades as a primary irrigant, other irrigants have been investigated as alternatives. This article reviews chlorhexidine as a canal irrigant, explores its different properties, and provides the dental practitioner with information to help make a more informed decision when choosing an irrigant.

Comparison of cytotoxicity and smear layer removal efficacy of triphala (an Indian ayurvedic herbal formulation) and 5.25% sodium hypochlorite as root canal irrigants: An in vitro study.

INTRODUCTIO: Healing potential of plants is an age-old idea that has recently attained renewed interest. Considering the ineffectiveness, potentially harmful effects, and safety concerns of commonly used synthetic irrigants, the herbal alternatives for endodontic usage might prove to be advantageous. AIM: The aim of this study was to evaluate the adequacy of smear layer removal and cytotoxicity potential of triphala in comparison to sodium hypochlorite. MATERIALS AND METHODS: The study was conducted in two parts: the first part of the study was cytotoxicity assessment studied using Alamar blue assay. L929 mouse fibroblasts were seeded in 96-well plates at a density of 5000 cells/well and treated with different concentrations of triphala and NaOCl for a period of 24 and 48 h. The percentage of cell viability was then quantified using an Alamar blue assay. The optical density was measured at 570 nm and compared with 620 nm, which was considered as a reference wavelength. The second part of the study was smear layer assessment at the coronal, middle, and apical third of twenty human premolar teeth using scanning electron microscope. RESULTS: The Alamar blue reagent cytotoxicity study suggested that triphala showed no cytotoxic properties against the normal mouse fibroblast cells whereas sodium hypochlorite showed a significant cytotoxic effect against the L929 cell lines with the IC50 concentration at 1.8%, respectively, after the treatment of 24 h of incubation at 37°C temperature. Triphala was as effective as sodium hypochlorite in smear layer removal in the coronal and middle third of the root, but sodium hypochlorite showed better smear layer removal in the apical third. CONCLUSION: Triphala can be considered as a superior irrigant with good antibacterial efficacy and least cytotoxicity potential compared to conventional hypochlorite irrigating agent and provide adequate clearing of smear layer in the coronal and middle third, and further studies are warranted to alter the properties of liquid to make it more cleansable in the apical third of the root.

Comparative assessment of time-related bioactive glass and calcium hydroxide effects on mechanical properties of human root dentin.

Suspensions of micro- or nanoparticulate SiO(2)-Na(2)O-CaO-P(2)O(5) bioactive glasses could potentially be used as dressings in traumatized front teeth with open apices as an alternative to Ca(OH)(2). These materials have a disinfecting capacity similar to Ca(OH)(2), but bear the advantage of bioactivity. However, because bioactive glasses initially act as alkaline biocides just as Ca(OH)(2) does, they may also negatively affect mechanical dentin properties over time. This was assessed in the current study using standardized human root dentin bars. Specimens were immersed in 1:20 (wt vol(-1)) suspensions of nanometric bioactive glass 45S5 or calcium hydroxide for 1, 10, or 30 days. Control specimens were immersed in pure saline for 30 days (n = 20 per group). Subsequently, modulus of elasticity (E) and flexural strength (FS) of the specimens were determined. Results were compared between groups using one-way anova and Scheffé's post-hoc test. Ca(OH)(2) caused a significant (P < 0.001) 35% drop in mean flexural strength values compared to the control treatment after 10 days. No further change was observed between 10 days and 30 days. Bioactive glass caused a 20% drop in mean flexural strength as compared to the control after 10 days. However, this difference did not reach statistical significance (P > 0.05). No effects of either material on dentin modulus of elasticity values were observed. It was concluded that the calcium hydroxide suspension affected the dentin more than the bioactive glass counterpart; however, the effect was self-limiting and probably restricted to superficial dentin layers, as suggested by the mere decrease in flexural strength but not in modulus of elasticity values.

Direct and Indirect Effect of Chlorhexidine on Survival of Stem Cells from the Apical Papilla and Its Neutralization.

INTRODUCTION: Several irrigants have been used for disinfection in regenerative endodontic procedures including chlorhexidine (CHX). In this context, the antibacterial properties of disinfectants are mainly in focus of research even though they may have an undesirable impact on the fate of stem cells. In this study, we hypothesized that CHX has both a direct effect when applied to stem cells of the apical papilla (SCAPs) and an indirect effect when SCAPs are exposed to dentin previously conditioned with CHX. METHODS: Cell toxicity was evaluated in vitro using the CellTox green fluorescence assay (Promega, Madison, WI) and CellTiter-Glo (Promega) after SCAPs were exposed directly to a dynamic concentration range of CHX; apical papilla explant cultures were stained with ApopTag (Merck Millipore, Billerica, MA) after culture with CHX. Furthermore, standardized slabs from human dentin were treated with CHX and consecutively rinsed in EDTA, L-α-lecithin (Sigma-Aldrich, St Louis, MO), or L-α-lecithin followed by EDTA. After that, SCAPs were cultured on the slabs for 5 days, and cellular viability was determined (indirect effect). Data were treated nonparametrically and analyzed using the Krukal-Wallis test (P ≤ .05). RESULTS: Direct exposure of SCAPs to CHX highly affected cell viability at concentrations above 10-3%, whereas lower concentrations had no adverse effect. During the initial 60 minutes, concentrations of 10-2% CHX or higher resulted in early pronounced toxicity with a maximum effect within 15 minutes after exposure. Likewise, CHX-conditioned dentin slabs were detrimental to SCAP survival; however, the deleterious effects were completely reversed by neutralization with L-α-lecithin. CONCLUSIONS: Chlorhexidine is toxic to SCAPs when applied directly or indirectly via conditioned dentin. If applied for a short time and neutralized by L-α-lecithin, it can be a gentle and cell-preserving disinfectant before endodontic regeneration.

Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA.

OBJECTIVE: The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC). MATERIAL AND METHODS: Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and » serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05. RESULTS: In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC. CONCLUSION: mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.

Influence of chlorine dioxide on cell death and cell cycle of human gingival fibroblasts.

OBJECTIVES: The effects of chlorine dioxide (ClO2), sodium hypochlorite (NaOCl), and hydrogen peroxide (H2O2) on cell death and the cell cycle of human gingival fibroblast (HGF) cells were examined. METHODS: The inhibition of HGF cell growth was evaluated using a Cell Counting Kit-8. The cell cycle was assessed with propidium iodide-stained cells (distribution of cells in G0/G1, S, and G2/M phases) using flow cytometry. The patterns of cell death (necrosis and apoptosis) were analyzed using flow cytometry with annexin V-FITC/PI staining. RESULTS: The lethal doses for 50% of the cells (LD50) of ClO2, NaOCl, and H2O2 were 0.16, 0.79, and 0.11 mM, respectively. All three dental disinfectants induced G0/G1 cell cycle arrest. H2O2 induced apoptosis at concentrations of 0.05 and 0.1 mM, while NaOCl and ClO2 did not induce significant apoptosis at any concentration examined. CONCLUSIONS: These results suggest that ClO2 is sufficient for use as a dental disinfectant compared with H2O2 or NaOCl.

Chlorhexidine gluconate in endodontics: an update review.

The major objective in root canal therapy is to disinfect the entire root canal system. This requires that the pulpal contents be eliminated as sources of infection. This goal may be accomplished using mechanical instrumentation and chemical irrigation, in conjunction with medication of the root canal between treatment sessions. Microorganisms and their by-products are considered to be the major cause of pulpal and periradicular pathosis. In order to reduce or eliminate bacteria, various irrigation solutions have been used during treatment. Chlorhexidine is a cationic molecule which can be used during treatment. It has a wide range antimicrobial activity. Furthermore, because of its cationic structure, chlorhexidine has a unique property named substantivity. The purpose of this paper is to review different aspects of the use of chlorhexidine gluconate in endodontics.

Assessment of toxicity and oxidative DNA damage of sodium hypochlorite, chitosan and propolis on fibroblast cells.

The objective of this study was to evaluate and compare the cytotoxicity and genotoxicity on human fibroblast cell lines of sodium hypochlorite (NaOCl), chitosan and propolis as root canal irrigating solutions. Human fibroblast cells were exposed to chitosan, propolis and NaOCl for 4 and 24 h. Cell viability was assessed by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide, and oxidative DNA damage was assessed by determination of 8-hydroxydeoxyguanosine (8-OHdG) level with an ELISA kit. The data of cell cytotoxicity were analysed statistically using a test of one-way analysis of variance at a significance level of p < 0.05. In the NaOCI group, the 8-OHdG level was higher than in the chitosan group, but there was no statistical difference when compared with the other groups (p < 0.05). It was determined that the irrigation solutions were cytotoxic, depending on the dose and time. NaOCl was the most toxic solution after both 4 and 24 h of exposure (p < 0.05). Chitosan and propolis may be alternatives to NaOCl for irrigation solutions, because they are both less toxic and produce less oxidative DNA damage.

Evaluation of chlorhexidine toxicity injected in the paw of mice and added to cultured l929 fibroblasts.

Because chlorhexidine (CHX) has been recommended as either an endodontic irrigant or root canal dressing, this study aimed to characterize, in vivo, the lesion induced by injections of CHX in the paw of mice at selected time intervals (24 and 48 hours and 7 and 14 days) and, in vitro, the mode of cell death, necrosis and/or apoptosis, and the cellular stress caused by exposition of cultured L929 fibroblasts to ascending concentrations of CHX for 24 hours. CHX injected in the subplantar space of the hind paw of mice induced severe toxic effects, as evidenced by necrotic changes in the epidermis, dermis, and subcutaneous tissue in association with reactive inflammatory response, particularly at higher concentrations. In addition, in cultured fibroblasts, CHX induced apoptosis at lower concentrations and necrosis at higher concentrations and increased expression of heat-shock protein 70, an indicator of cellular stress. Taken together, these findings suggest that CHX may have an unfavorable effect on the resolution of apical periodontitis.

Assessment of genotoxicity of 14 chemical agents used in dental practice: ability to induce chromosome aberrations in Syrian hamster embryo cells.

To assess the genotoxicity of 14 chemical agents used as locally applied agents in dental practice, the ability of these agents to elicit chromosome aberrations was examined using Syrian hamster embryo (SHE) cells. Chromosome aberrations in SHE cells were induced by treatment with three of eight chemical agents used as endodontic medicaments, i.e. ethylenediaminetetraacetic acid (EDTA), formocresol (a mixture of formalin and tricresol), and sodium arsenite. The other five chemical agents, i.e. chloramphenicol, p-chlorophenol, p-phenolsulfonic acid, sodium hypochlorite, and tetracycline hydrochloride exhibited a negative response for chromosome aberrations. Assessment of three dyes used for disclosing dental plaque showed chromosome aberrations induced by basic fuchsin but not by acid fuchsin and erythrosine B. Three local anesthetics, lidocaine hydrochloride, prilocaine hydrochloride, and procaine hydrochloride, were negative for chromosome aberrations. Among the ten chemical agents that exhibited a negative response in the assay, p-chlorophenol, sodium hypochlorite, and erythrosine B induced chromosome aberrations in SHE cells when treated in the presence of exogenous metabolic activation. The percentages of cells with polyploidy or endoreduplication were enhanced by formocresol, sodium arsenite, p-chlorophenol, p-phenolsulfonic acid, sodium hypochlorite, erythrosine B, prilocaine hydrochloride, and procaine hydrochloride in the absence or presence of exogenous metabolic activation. Our results indicate that the chemical agents that had a positive response in the present study are potentially genotoxic to mammalian cells.