Comparative analysis of the effects of ozone and ultrasound on Streptococcus mutans

Streptococcus mutans is one of the principal pathogens of the human oral habitat, being one of the principal etiological agents of carious lesions. Ozone is a powerful oxidant, it has the ability to inactivate microorganisms in general, and ultrasound is an acoustic system generated through a piezoelectric crystal that also presents microbicidal effects. In the present study, a comparative analysis was made of the damage caused to Streptococcus mutans in vitro by ultrasound and ozone, through scanning electron microscopy (SEM) and atomic force microscopy (AF) analysis. In addition, flow cytometry was used to determine microbial viability and the formation of Reactive Oxygen Species (ROS) after the application of different techniques. The data obtained by means of micro- scopic analysis reveal that both ozone and ultrasound produce morphological alterations in bacteria, which become rod-shaped organisms. In addition to this deformation, on the microbial surface it was possible to identify crater-like impressions. In contrast, the irregularity of protuber- ances on the surface of the microbial wall was only detected when ozone was employed. Regarding the formation of ROS, it was observed that that ozone induces a significant growth (p < .05) of these molecules, while ultrasound does not present this effect. Ozone and ultrasound present micro- bicidal effects, however, ozone is more efficient.

Analysis of damage on the Streptococcus mutans immersed in ozonated water: preliminary study for application as mouth rinse

Ozonated water has been demonstrated to induce significant results in terms of the elimination ofmicroorganisms. The present study assessed the damage toStreptococcusmutansafter exposureto ozonated water; the ozone generator was adjusted to provide an outlet concentration of60 mg/L, the samples were submitted to different ozonation times 1, 2, 4, 6, and 10 mi. Scanningelectron microscopy and atomic force images were obtained to identify damage to the bacteria,followed by reactive oxygen species (ROS) evaluation and microbial viability. The results showed asignificant reduction in viability and the images evidenced the generation of gaps on themicrobial wall and surface layer alterations. Ozone can induce significant damage toS.mutans,thus suggesting that the use of ozonated water to prevent carious lesion formation is extremelypromising.

Antiproliferative and antiangiogenic effect of Amblyomma sculptum (Acari: Ixodidae) crude saliva in endothelial cells in vitro

Neovascularization, a process that includes vasculogenesis and angiogenesis, may be a physiological or pathologic event, but in any cases the phenomenon is related to the formation of vascular net and sprouting of endothelial cells from preexisting blood vessel. The tumor environment, which counts on the tumor cell proliferation, is plenty of proangiogenic factors, such as angiogenin, TGF (a and ß), FGF, VEGF, all of them playing a crucial role in angiogenesis, an important hallmark of cancer frequently related to a poor prognosis. Therefore, therapies focusing the inhibition of cancer neovasculogenesis have become an interesting strategy for the development of antitumor therapies. In this work, we investigate the effect of tick saliva on the human endothelial cells, in order to understand its inhibitory effects on angiogenesis. To this end, the HUVEC cells were used as model of angiogenesis in vitro and the anti-proliferative, anti-migratory, cytotoxicity was evaluated. Our data depicts that saliva impairs cell development by causing structural changes while precludes cell proliferation and migration, that are crucial events related to angiogenesis. Aiming the identification of the bioactive components related to antiangiogenic activity, saliva was analyzed through the Mass Spectrometry and among all molecules identified, disintegrins and cathepsin L seems to be primarily responsible for the antiangiogenic effects of saliva.

Analysis of damage on the Streptococcus mutans immersed in ozonated water: preliminary study for application as mouth rinse

Ozonated water has been demonstrated to induce significant results in terms of the elimination ofmicroorganisms. The present study assessed the damage toStreptococcusmutansafter exposureto ozonated water; the ozone generator was adjusted to provide an outlet concentration of60 mg/L, the samples were submitted to different ozonation times 1, 2, 4, 6, and 10 mi. Scanningelectron microscopy and atomic force images were obtained to identify damage to the bacteria,followed by reactive oxygen species (ROS) evaluation and microbial viability. The results showed asignificant reduction in viability and the images evidenced the generation of gaps on themicrobial wall and surface layer alterations. Ozone can induce significant damage toS.mutans,thus suggesting that the use of ozonated water to prevent carious lesion formation is extremelypromising.

Antiproliferative and antiangiogenic effect of Amblyomma sculptum (Acari: Ixodidae) crude saliva in endothelial cells in vitro

Neovascularization, a process that includes vasculogenesis and angiogenesis, may be a physiological or pathologic event, but in any cases the phenomenon is related to the formation of vascular net and sprouting of endothelial cells from preexisting blood vessel. The tumor environment, which counts on the tumor cell proliferation, is plenty of proangiogenic factors, such as angiogenin, TGF (a and ß), FGF, VEGF, all of them playing a crucial role in angiogenesis, an important hallmark of cancer frequently related to a poor prognosis. Therefore, therapies focusing the inhibition of cancer neovasculogenesis have become an interesting strategy for the development of antitumor therapies. In this work, we investigate the effect of tick saliva on the human endothelial cells, in order to understand its inhibitory effects on angiogenesis. To this end, the HUVEC cells were used as model of angiogenesis in vitro and the anti-proliferative, anti-migratory, cytotoxicity was evaluated. Our data depicts that saliva impairs cell development by causing structural changes while precludes cell proliferation and migration, that are crucial events related to angiogenesis. Aiming the identification of the bioactive components related to antiangiogenic activity, saliva was analyzed through the Mass Spectrometry and among all molecules identified, disintegrins and cathepsin L seems to be primarily responsible for the antiangiogenic effects of saliva.

The Roles of ROS in Cancer Heterogeneity and Therapy.

Cancer comprises a group of heterogeneous diseases encompassing high rates of morbidity and mortality. Heterogeneity, which is a hallmark of cancer, is one of the main factors related to resistance to chemotherapeutic agents leading to poor prognosis. Heterogeneity is profoundly affected by increasing levels of ROS. Under low concentrations, ROS may function as signaling molecules favoring tumorigenesis and heterogeneity, while under high ROS concentrations, these species may work as cancer modulators due to their deleterious, genotoxic or even proapoptotic effect on cancer cells. This double-edged sword effect represented by ROS relies on their ability to cause genetic and epigenetic modifications in DNA structure. Antitumor therapeutic approaches may use molecules that prevent the ROS formation precluding carcinogenesis or use chemical agents that promote a sudden increase of ROS causing considerable oxidative stress inside tumor mass. Therefore, herein, we review what ROS are and how they are produced in normal and in cancer cells while providing an argumentative discussion about their role in cancer pathophysiology. We also describe the various sources of ROS in cancer and their role in tumor heterogeneity. Further, we also discuss some therapeutic strategies from the current landscape of cancer heterogeneity, ROS modulation, or ROS production.

The roles of ROS in cancer heterogeneity and therapy

Cancer comprises a group of heterogeneous diseases encompassing high rates of morbidity and mortality. Heterogeneity, which is a hallmark of cancer, is one of the main factors related to resistance to chemotherapeutic agents leading to poor prognosis. Heterogeneity is profoundly affected by increasing levels of ROS. Under low concentrations, ROS may function as signaling molecules favoring tumorigenesis and heterogeneity, while under high ROS concentrations, these species may work as cancer modulators due to their deleterious, genotoxic or even proapoptotic effect on cancer cells. This double-edged sword effect represented by ROS relies on their ability to cause genetic and epigenetic modifications in DNA structure. Antitumor therapeutic approaches may use molecules that prevent the ROS formation precluding carcinogenesis or use chemical agents that promote a sudden increase of ROS causing considerable oxidative stress inside tumor mass. Therefore, herein, we review what ROS are and how they are produced in normal and in cancer cells while providing an argumentative discussion about their role in cancer pathophysiology. We also describe the various sources of ROS in cancer and their role in tumor heterogeneity. Further, we also discuss some therapeutic strategies from the current landscape of cancer heterogeneity, ROS modulation, or ROS production.

Apoptotic effect of eugenol envolves G2/M phase abrogation accompanied by mitochondrial damage and clastogenic effect on cancer cell in vitro.

BACKGROUND: Eugenol (EUG) is a major phenolic compound present in clove whose anti-cancer properties have been demonstrated previously. These anti-cancer properties may involves the modulation of different mechanisms, including α-estrogen receptor (αER) in luminal breast cancer cells, COX-2 inhibition in melanoma cells or p53 and caspase-3 activation in colon cancer cells. HYPOTHESIS: EUG promotes a burst in ROS production causing cell-cycle perturbations, mitochondria toxicity and clastogenesis triggering apoptosis in melanoma breast- and cervix-cancer cells in vitro. METHODS: Morphological changes were evaluated through the light- and electronic- microscopy. Cell-cycle, ROS, PCNA and Apoptosis was detected by flow cytometry and clastogenicity was evaluated by Comet-assay. RESULTS: The results obtained herein pointed out that EUG promotes, increasing ROS production leading to abrogation of G2/M of phase of cell-cycle, and consecutively, clastogenesis in vitro. In addition, EUG induces Proliferation Cell Nuclear Antigen (PCNA) downregulation and decreasing in mitochondria potential (ΔΨm). Of note, a Bax up-regulation was also observed on cells treated with EUG. All of these findings cooperate in order to induce apoptosis in cancer cells. CONCLUSION: These promising results presented herein shed new light on the mechanisms of action of EUG suggesting a possible applicability of this phenylpropanoid as adjuvant in anti-cancer therapy.