Ozonated Water Treatment Research Articles

Evaluation of oxygenated and ozonated nanobubble water treatments for dollar spot suppression in seashore paspalum

AbstractDollar spot (caused by Clarireedia spp.) is the most commonly occurring turfgrass disease on golf courses in North America, and current disease control programs rely on frequent fungicide applications. The escalating occurrence of fungicide resistance in Clarireedia spp. populations, coupled with the reduction of the annual kilograms active ingredient applied per hectare for some fungicides, emphasizes the need for alternative management strategies. The use of oxygenated or ozonated water treatments has been effective as a component of an overall plant disease management strategy. In field and growth chamber‐controlled environment trials, the impacts of oxygenated and ozonated nanobubble water treatments were evaluated for turf quality and dollar spot control in seashore paspalum (Paspalum vaginatum Swartz). Despite generating relatively high levels of dissolved oxygen (40 mg L−1) or ozone (ca. 8 mg L−1) in water treatments through nanobubble aeration in all trials, these treatments did not cause damage to seashore paspalum tissues, but were unsuccessful in controlling dollar spot. Additionally, tests comparing two different application methods (soil drench versus foliar spray) for all treatments suggested that the application method had no effect on treatment efficacy. Overall, oxygenated and ozonated nanobubble water treatments did not adversely affect seashore paspalum turf quality and were ineffective in suppressing dollar spot in field and growth chamber trials.

A Meta-Analysis on the Antimicrobial Effectiveness of Ozonated Water Treatments for Fresh Produce Washing-Effect of Ozonation Methods.

Due to the lack of a pathogen-killing process, foodborne outbreaks from contaminated fresh produce have been increasing worldwide. Hence, it is increasingly recognized that the washing step with sanitizers is important to control microbial contamination. Ozonated water is suggested as a substitute for chlorine-based sanitizers, addressing concerns about the effectiveness and environmental impact of chlorine-based sanitizers. However, using ozone as a sanitizer in the fresh produce washing process is still challenging because of its unstable and inconsistent antimicrobial effectiveness under various testing conditions. A meta-analysis was focused on the comparison of antimicrobial effectiveness between different ozonation methods commonly adopted in laboratory settings, including stationary pre-ozonated water, agitated pre-ozonated water, and sparging. The meta-analysis showed that the sparging method results in the highest microbial log reduction compared to other methods. We further developed meta-regression models based on three ozonation methods to identify key processing variables influencing the antimicrobial effectiveness of ozonated water. Attempts were made to link key processing variables to ozone stability and the mass transport phenomena involved in the washing process. This research will contribute to designing and developing a washing process to increase fresh produce safety by identifying key factors in each ozonation method and facilitate interlaboratory comparison studies.

The quality of chili (Capsicum annum L.) during storage after ozone water treatment

Abstract Chili is one of the main agricultural commodities. During the peak harvest season, there is an increase in chili production, so it is necessary to apply post-harvest handling technology to maintain the quality of chili, one of which is through the application of ozone technology. The object of this study is to evaluate the quality of chili (Capsicum annum L.) during storage after ozone water treatment. The material used in this study was curly chilies; the treatments used 2 types of storage temperatures (28 °C and 3 °C) and 2 types of washing processes (soaking and spraying). Observations were made on days 0, 7, and 14. Parameters observed included weight loss, moisture content, total bacteria, and total yeast. The results showed that the total bacteria value on day 7 decreased to 1 x 105 CFU/g. The total yeast of chilies with ozone treatment showed a significant reduction in total yeast compared to the control treatment. The weight loss of control chilies and washed chilies stored at room temperature after being stored for 14 days was in the range of 29.64% to 58.94%. The highest moisture content was obtained by spray treatment and stored at 3 °C.

СНИЖЕНИЕ ЖЁСТКОСТИ ОРОСИТЕЛЬНОЙ ВОДЫ ОЗОНИРОВАНИЕМ

Abstract. The article presents the research on the effect of ozone treatment on the hardness of irrigation water. The effect of hard water on plant growth and development has been analyzed. It has been revealed that excessive magnesium and calcium ions in irrigation water lead to the suppression of agricultural crops and a decrease in their yield. To conduct laboratory studies, water was taken from a natural reservoir located near the village of Gvazda, Buturlinovsky District, Voronezh Region, which is used by local farmers to irrigate agricultural land. The sample was divided into two parts, one of which was subjected to ozone treatment, and the second was a control variant. Laboratory studies have established that the total water hardness in the natural reservoir was 21 dH, which corresponds to 7.5 mg eq / l. This level of hardness is excessive and will negatively affect plants. Ozonation at an average ozone concentration in the ozone-air mixture of 4.67 mg/m3 allowed to reduce the total water hardness to 14 dH, which is comparable to 5 mg eq/l. Intensive water softening occurred at ozone treatment doses of up to 840 mg min/m3. Consequently, ozonation reduces the content of polyvalent metal ions, which in high concentrations have a negative effect on plants. Modern compact, energy-efficient, high-performance ozonizers can be easily integrated into the design of sprinkler machines. Softer water will have a positive effect on the efficiency of irrigation and the reliability of irrigation systems. In addition, ozone water treatment will have a bactericidal and disinfectant effect, which will prevent the development of various diseases and pests.

From Waste to Resource: Evaluating Biomass Residues as Ozone-Catalyst Precursors for the Removal of Recalcitrant Water Pollutants

Five different biomass wastes—orange peel, coffee grounds, cork, almond shell, and peanut shell—were transformed into biochars (BCs) or activated carbons (ACs) to serve as adsorbents and/or ozone catalysts for the removal of recalcitrant water treatment products. Oxalic acid (OXL) was used as a model pollutant due to its known refractory character towards ozone. The obtained materials were characterized by different techniques, namely thermogravimetric analysis, specific surface area measurement by nitrogen adsorption, and elemental analysis. In adsorption experiments, BCs generally outperformed ACs, except for cork-derived materials. Orange peel BC revealed the highest adsorption capacity (Qe = 40 mg g−1), while almond shell BC showed the best cost–benefit ratio at €0.0096 per mg of OXL adsorbed. In terms of catalytic ozonation, only ACs made from cork and coffee grounds presented significant catalytic activity, achieving pollutant removal rates of 72 and 64%, respectively. Among these materials, ACs made from coffee grounds reveal the best cost/benefit ratio with €0.02 per mg of OXL degraded. Despite the cost analysis showing that these materials are not the cheapest options, other aspects rather than the price alone must be considered in the decision-making process for implementation. This study highlights the promising role of biomass wastes as precursors for efficient and eco-friendly water treatment processes, whether as adsorbents following ozone water treatment or as catalysts in the ozonation reaction itself.

Green control for inhibiting Rhizopus oryzae growth by stress factors in forage grass factory.

The forage grass factory could break through the restrictions of land resources, region and climate to achieve efficient production throughout the year by accurate and intelligent management. However, due to its closed environment, mold outbreaks in the forage grass factory were severe, significantly affecting barley production. In this study, 9 contaminated barley tissues were collected and 45 strains were isolated in forage grass factory. After ITS sequencing, 45 strains were all identified as Rhizopus oryzae. Through stress factor assays, R. oryzae growth was seriously hindered by low concentration of sodium nitrate, high pH value and ozone water treatment. High pH and ozone water affected growth mainly by altering membrane integrity of R. oryzae. Sodium nitrate inhibited the growth of R. oryzae mainly by affecting the amount of sporulation. Low concentration of sodium nitrate and ozone water did not affect the growth of barley. High concentrations of sodium nitrate (100 mM) and pH values (8-8.5) inhibited barley growth. Among them, ozone water had the most obvious inhibition effect on R. oryzae. Large-scale ozone water treatment in the forage grass factory had also played a role in restoring barley production. Taken together, the green techonology to control mold disease and maintain the safety of forage through different physicochemical methods was selected, which was of considerable application value in animal husbandry.

Ozone Treatment as a Sustainable Alternative for Suppressing Blue Mold in Mandarins and Extending Shelf Life

Citrus fruits, particularly mandarins, are highly valued globally for their nutritional benefits and versatile culinary uses. However, the challenge of post-harvest decay, primarily due to blue mold (Penicillium italicum) infections, results in significant food losses and necessitates effective preservation strategies. Traditional methods often rely on fungicides, raising concerns about chemical residues and environmental impact. This study investigates the efficacy of ozone as an alternative approach to controlling blue mold in mandarins. Various gaseous ozone treatments were tested, including single, double, and triple treatments, with durations ranging from 10 to 60 min and concentrations from 3.3 to 20 ppm. Additionally, ozonated water treatments were evaluated with concentrations of 2, 4, and 6 ppm. To simulate a realistic infestation scenario, mandarins were artificially infected with P. italicum spores before undergoing both gaseous ozone and ozonated water treatments. The storage conditions for the mandarins were meticulously controlled, maintaining a humidity level of 50–60% and a temperature range of 10–12 °C. Each fruit was analyzed, and the presence of P. italicum infection was determined two and three weeks after the ozonation. Results indicated that ozone treatments significantly reduced mold growth, with gaseous ozone demonstrating efficacy rates up to 97.5% and ozonated water treatments achieving preservation rates between 95% and 97%. These results underscore ozone’s potential as a safe, efficient, and sustainable alternative to conventional fungicides, offering promising solutions for extending the shelf life of mandarins. Further research is recommended to optimize ozone treatment parameters, assess long-term effects on fruit quality and nutritional content, and refine application techniques to harness ozone’s potential in citrus fruit preservation fully. This approach not only addresses food security challenges but also aligns with global efforts to reduce chemical inputs in agriculture and promote environmentally sustainable practices.

Origin of the Activity of Electrochemical Ozone Production Over Rutile PbO2 Surfaces.

Ozonation water treatment technology has attracted increasing attention due to its environmental benign and high efficiency. Rutile PbO2 is a promising anode material for electrochemical ozone production (EOP). However, the reaction mechanism underlying ozone production catalyzed by PbO2 was rarely studied and not well-understood, which was in part due to the overlook of the electrochemistry-driven formation of oxygen vacancy (OV) of PbO2. Herein, we unrevealed the origin of the EOP activity of PbO2 starting from the electrochemical surface state analysis using density functional theory (DFT) calculations, activity analysis, and catalytic volcano modeling. Interestingly, we found that under experimental EOP potential (i. e., a potential around 2.2 V vs. reversible hydrogen electrode), OV can still be generated easily on PbO2 surfaces. Our subsequent kinetic and thermodynamic analyses show that these OV sites on PbO2 surfaces are highly active for the EOP reaction through an interesting atomic oxygen (O*)-O2 coupled mechanism. In particular, rutile PbO2(101) with the "in-situ" generated OV exhibited superior EOP activities, outperforming the (111) and (110) surfaces. Finally, by catalytic volcano modeling, we found that PbO2 is close to the theoretical optimum of the reaction, suggesting a superior EOP performance of rutile PbO2. All these analyses are in good agreement with previous experimental observations in terms of EOP overpotentials. This study provides the first volcano model to explain why rutile PbO2 is among the best metal oxide materials for EOP and provides new design guidelines for this rarely studied but industrially promising reaction.

Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the USU Dental and Oral Hospital

Background The application of disinfectants on dental instruments is one way to prevent cross-infection. Cross infection can occur due to microorganisms found in blood, saliva and dental plaque which can contaminate the instruments used by dental health workers. Thus, indirect contact transmission of pathogenic microorganisms occurs from instruments that have been contaminated by dental health workers. Ozone water and 4.8% chloroxylenol are disinfecting agents used to disinfect medical instruments. This study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October-December 2022. Methods The samples used were mandibular molar pliers that have been used in tooth extraction procedures. This study was experimental and used three sample groups, where each group consisted of 10 tooth extraction instruments. The treatment group used ozone water and 4.8% chloroxylenol and the negative control group was cleaned with distilled water. The test effectiveness in this study used bacteria colony counter using the scatter cup method. Data were analyzed using the Kruskal Wallis and Mann-Whitney U tests. Results The results of the data analysis showed a p-value ≤0.001, which means that there was a significant difference in the disinfection using ozone water and 4.8% chloroxylenol on the number of bacterial colonies on dental extraction instruments. The results of this study show that the average number of bacterial colonies formed in the ozone water treatment group was 4.00 ± 4.32, 16.00 ± 6.65 in the 4.8% chloroxylenol treatment group, and 217.50 ± 39.24 in the negative control group (Aquadest). Conclusions From this study it can be said that ozone water is more effective in disinfecting dental extraction instruments than 4.8% chloroxylenol.

Application Of Advanced Oxidation Process Based on Ozone in Water Treatment

Wastewater treatment is an effective way to address water scarcity. Advanced oxidation processes (AOPs) are a wastewater treatment technology that is currently attracting a lot of attention, and it is a technology that uses physical or chemical pathways such as ozone, ultraviolet light, and the Fenton reaction to generate free radicals and then treats pathogens and organic pollutants by oxidation of the free radicals.Ozone is very strong oxidising agent and it has a powerful ability to break down wide range of organic and inorganic pollutants. However its usage in the wastewater treatment is still not very popular due to several drawbacks it posses, as it not possible to achieve the goal of wastewater treatment by ozone alone due to its mass transfer limitations, slow reaction rates, non specific for certain under low reaction rates pollutants select,may costs relatively higher thanother treatment processes etc. By combining ozone with H2O2, UV light, and catalysts, the efficiency of the wastewater treatment process will be increased because it will help make more hydroxyl radicals. This review discusses various ozone-based AOPs such as ozone/UV, ozone/hydrogen peroxide, and catalytic ozonation, focusing on their mechanisms and applications in wastewater treatment.

TECHNOLOGIES WITH OZONE IN HEMODIALYSIS

This study aimed to verifying the impacts of technologies with ozone in water treatment and disinfection of hemodialysis water systems, replacing the conventional chemical processes. Through scientific research developed on the subject, the study conducted a comparative analysis between the conventional technologies and technologies using ozone, in order to evaluate positive aspects and fragilities; and demonstrate it’s contribution to the practice of sustainability in economic, social and environmental aspects. The results of these analyzes demonstrated that the technologies with ozone are a safe and effective method, which results in the improvement of hemodialysis; giving quality and sustainability to the process.

Methods for SARS-CoV-2 hospital disinfection, in vitro observations

IntroductionEscalation of chemical disinfection during the COVID-19 pandemic raised occupational hazard concerns. Alternative and potentially safer methods such as ultraviolet-C (UVC) irradiation and ozone are desirable, notwithstanding the lack of standardized criteria for their use in the healthcare environment. AimCompare the virucidal activity of 70% ethanol, sodium dichloroisocyanurate (NaDCC), chlorhexidine, ozonated water, UVC-222 nm, UVC-254 nm against three SARS-CoV-2 variants of concern cultured in vitro. MethodsInactivation of three SARS‐CoV‐2 variants (alpha, beta, gamma) by the following chemical methods was tested: ethanol 70%, NaDCC (100 ppm, 500 ppm, 1000 ppm), chlorhexidine (2%, 1% and 0.5%), ozonated water 7 ppm. For irradiation, a je2Care 222nm UVC Lamp was compared to a Sylvania G15 UV254 nm lamp on SARS-CoV-2 variants. ResultsViral inactivation by >3 log was achieved with ethanol, NaDCC and chlorhexidine. The minor virucidal effect of ozonated water was <1 log. Virus treatment with UVC-254 nm reduced viral activity by 1-5 logs with higher inactivation after exposure for 3 minutes compared to 6 seconds. For all three variants, under equivalent conditions, exposure to UVC-222 nm did not achieve time-dependent inactivation as was observed with treatment with UVC-254 nm. ConclusionThe virucidal activity on replication-competent SARS-CoV-2 by conventional chemical methods, including chlorhexidine at concentrations as low as 0.5%, was not matched by UVC irradiation, and to an even lesser extent by ozonated water treatment.

Therapeutic potential of ozone water treatment in alleviating atopic dermatitis symptoms in mouse models: Exploring its bactericidal and direct anti-inflammatory properties

Currently, ozone water is utilized for antibacterial and antiviral purposes without any reported safety concerns. Therefore, ozone water may have clinical applications in treating staphylococcal-specific cutaneous diseases, such as atopic dermatitis (AD) and pyoderma. This study aimed to verify the bactericidal effects of ozone water at different concentrations (3 and 11 mg/L) against staphylococcal species in vitro, as well as evaluate the anti-inflammatory effects of ozone water in a mouse model of AD and pyoderma. Initially, the bactericidal properties of several concentrations of ozone water were confirmed with Staphylococcus aureus and methicillin-resistant S. pseudintermedius. Both 3 and 11 mg/L of ozone water exhibited a significant bactericidal effect against staphylococci at less than 100 times dilution. We next examined the cellular cytotoxicity and cytokine production (Interleukin (IL)-6 and IL-8) induced by S. pseudintermedius pre-treated with ozone water, and our findings indicated that cytotoxicity and cytokine production induced by staphylococci were significantly inhibited after ozone water pre-treatment. In vivo experiments showed that ozone water-pre-treated S. pseudintermedius significantly inhibited the development of pyoderma in mice; however, limited effects were observed in a therapeutic setting. Interestingly, ozone water at concentrations of 3 and 11 mg/L exhibits dual bactericidal and anti-inflammatory effects in mice with AD. This observation was corroborated by the significant inhibition of cytokine production in interferon-γ/tumor necrosis factor-stimulated human epidermal keratinocyte cells exposed to ozone in vitro. These findings indicate that administering ozone can be a novel therapeutic approach for managing allergic skin diseases, such as AD.

Pesticide Residues and Berry Microbiome after Ozonated Water Washing in Table Grape Storage.

Nowadays, different systems for reducing pesticides in table grapes are being tested at different production stages either in the field or in postharvest. The present study tested ozonated water treatments at the beginning of the cold storage of the Princess® seedless table grape variety to reduce the residue contents of some pesticides and to evaluate their effect on gray mold and the berry microbiome. An ozone generator capable of producing an ozone concentration ranging from 18 to 65 Nm3 was utilized for obtaining three ozone concentration levels in water: 3, 5 and 10 mg/L. Ozonated water was placed in a 70 L plastic box where 500 g grape samples closed in perforated plastic clamshell containers were immersed utilizing two washing times (5 and 10 min). Overall, six ozonated water treatments were tested. After the ozonated water treatments, all samples were stored for 30 days at 2 °C and 95% relative humidity to simulate commercial practices. The pesticide residue contents were determined before the ozonated water treatments (T0) and 30 days after the cold storage (T1). The treatments with ozonated water washing reduced the pesticide residues up to 100%, while the SO2 control treatment reduced the pesticide residues ranging from 20.7 to 60.7%. Using 3 mg/L ozonated water to wash grapes for 5 min represented the optimal degradation conditions for all of the analyzed pesticides, except for fludioxonil, which degraded better with a washing time of 10 min. The ozone treatments did not significantly reduce the gray mold and the fungal and bacterial microbiome, while a relevant reduction was observed in the yeast population.

Construction of triphase interface for catalytic ozonation of polluted water

The utilization efficiency of ozone determines the cost of catalytic ozonation in water treatment. Herein, a triphase catalytic system was constructed by aerating ozone through a CeO2 loaded Al2O3 ceramic membrane (CeO2-CM) for disinfection and antibiotic degradation. Ozone aeration and a packed catalyst system (CeO2-Packing) were set as the controls. Results showed that CeO2-CM reduced the ozone escape by 34.6%–56.2%. The ozone utilization capacity of CeO2-CM for E. coli inactivation was 33.1% and 33.8% higher than those of CeO2-Packing and ozone aeration, respectively. The ozone utilization capacity of CeO2-CM for sulfamethoxazole degradation was 88.5% and 183.1% higher than those of CeO2-Packing and ozone aeration, respectively. CeO2-CM, with the lowest ozone escape and highest ozone utilization efficiency, significantly enhanced the performance of catalytic ozonation in disinfection and antibiotic degradation. This work proposes a feasible strategy for minimizing ozone consumption in water treatment.

The disinfection effectiveness of ozone water and 4.8% chloroxylenol against the number of bacterial colonies in dental extraction instruments at the USU Dental and Oral Hospital in October-December 2022

Background: The application of disinfectants on dental instruments is one way to prevent cross-infection. Cross infection can occur due to microorganisms found in blood, saliva and dental plaque which can contaminate the instruments used by dental health workers. Thus, indirect contact transmission of pathogenic microorganisms occurs from instruments that have been contaminated by dental health workers. Ozone water and 4.8% chloroxylenol are disinfecting agents used to disinfect medical instruments. This study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October-December 2022. Methods: The samples used were mandibular molar pliers that have been used in tooth extraction procedures. This study was experimental and used three sample groups, where each group consisted of 10 tooth extraction instruments. The treatment group used ozone water and 4.8% chloroxylenol and the negative control group was cleaned with distilled water. The test effectiveness in this study used bacteria colony counter using the scatter cup method. Data were analyzed using the Kruskal Wallis and Mann-Whitney U tests. Results: The results of the data analysis showed a p-value ≤0.001, which means that there was a significant difference in the disinfection using ozone water and 4.8% chloroxylenol on the number of bacterial colonies on dental extraction instruments. The results of this study show that the average number of bacterial colonies formed in the ozone water treatment group was 4.00 ± 4.32, 16.00 ± 6.65 in the 4.8% chloroxylenol treatment group, and 217.50 ± 39.24 in the negative control group (Aquadest). Conclusions: From this study it can be said that ozone water is more effective in disinfecting dental extraction instruments than 4.8% chloroxylenol.

Optimum conditions of the treatment of organic matters in gray water by Ozone

Wastewater treatment processes are critical and economical to improve the quality, remove the more significant part of the contaminants from wastewater and solve the water crisis, which may provide new water sources. To conserve water, the gray water (GW) must be recycled. The use of Ozone in water treatment is a chemical treatment based on the infusion of Ozone into water or wastewater. In this study, the best conditions for treating gray water with Ozone were investigated to achieve the best removal of organic matters represented by chemical oxygen demand (COD), Biochemical oxygen demand (BOD₅), Total organic carbon (TOC), Oils and Greases. It was experimentally concluded that the ozone treatment is positively affected by the increase in pH, the ozone concentration and the time of exposure to Ozone. The ideal conditions for ozone treatment were determined according to the quality of the resulting water and not according to the removal values of organic materials. These conditions are pH 8, ozone concentration of 40mg/l and ozone exposure time of 30 min. Keywords: Gray water; Ozonation; Water Treatment; COD.

Resist Removal Using Laser Irradiation Combined with Ozone Water Treatment

Resist Removal Using Laser Irradiation Combined with Ozone Water Treatment

Effects of Cold Plasma and Ozone Water Treatment on Micronutrient Solubility

Cold plasma and ozone sanitation of irrigation solutions can oxidize both microbes and non-target micronutrients because their high oxidation-reduction potential (ORP) is a non-selective mode of action. The objective of this study was to evaluate the effects of cold plasma and ozone treatment on oxidation of iron and manganese in nutrient solutions containing one of four iron chelates (iron-ethylenediaminetetraacetic acid (Fe-EDTA), iron-diethylenetriaminepentaacetic acid (Fe-DTPA), iron-ethylenediamine-N,N′-bis(2-hydroxyphenylacetic acid) (Fe-EDDHA), and hydroxybenzyl ethylenediamine (Fe-HBED)). Nutrient solutions were recirculated through the cold plasma or ozone system until the ORP reached 700 mV. The concentrations of total dissolved iron, manganese, and chelated iron were measured before and after passing through the treatment systems. Both cold plasma and ozone oxidized chelates and decreased the solubility of iron and manganese. Cold plasma and ozone had similar effects on micronutrients, pH, electrical conductivity, and dissolved oxygen at a standardized target ORP of 700 mV. Fe-EDTA was the most resistant chelate to oxidation. With Fe-EDTA, ORP increased more quickly, and the concentration of chelated Fe decreased less with the increasing ORP over time compared with Fe-DTPA, Fe-EDDHA, and Fe-HBED. The concentration of chelated Fe decreased by up to 80% for EDDHA at 700 mV compared with a 20% decrease for EDTA. The concentration of Mn decreased by up to 85% at 700 mV. The design of water treatment with cold plasma or ozone therefore requires consideration of secondary effects on micronutrients. The treatment dosage, flow rate, and nutrient solution at a particular grower operation are likely to affect the quantity of micronutrient fertilizer that needs to be supplemented following treatment. Use of Fe-EDTA is one strategy to reduce the loss of iron and increase residual ORP that is available for sanitation.

Ozonated Water Inhibits Hepatocellular Carcinoma Invasion and Metastasis by Regulating the HMGB1/NF-κB/STAT3 Signaling Pathway.

Hepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide. High-mobility group box 1 (HMGB1), a highly conserved chromosome protein, is considered as a potential therapeutic target and novel biomarker because of its regulation in the proliferation and metastasis of HCC. Ozone has been shown to be beneficial in the treatment of cancer. The objective of this study was to explore the effects and molecular mechanism of ozonated water on the proliferation, migration, and invasion of BEL7402 HCC cells. We assessed cell proliferation using a CCK-8 assay kit and flow cytometry; we performed wound healing and transwell assays to evaluate the effects of ozonated water treatment on cell invasion and migration. We determined reactive oxygen species (ROS) values by flow cytometry and used ELISAs to detect cytokines HMGB1, IL-6, and TNF-α. In addition, we assessed mRNA and protein cytokine expressions using RT-qPCR and Western blot. Ozonated water decreased the viability of the HCC cells; the IC50 of ozonated water at 24 h was approximately 1.5 μg/mL. Compared with control groups, ozone treated cells revealed reduced mobility on wound healing assays and decreased invasion in transwell assays. HMGB1, IL-6, and TNF-α cytokines were found at lower levels in ozone treated cells than in control cells. Ozonated water-induced ROS accumulation. Likewise, the expressions of phosphorylated nuclear factor Kappa B (NF-κB), p65, NF-κB, P-STAT3, IL-6, JAK2, Slug, Twist, vimentin, MMP-2, MMP-9, and HMGB1 were decreased in the treated cells. Our findings suggest that ozonated water inhibits the proliferation, invasion, and metastasis of HCC cells via regulation of the HMGB1/NF-κB/STAT3 signaling pathway.