Table 2 Electrolyzed water used against cultured oral bacteria

1

Okamura T., et al. 2019 [29]

Nihon University, Tokyo, Japan

In vitro

To assess antimicrobial and noxious effects of acid/alkaline electrolyzed FW compared with NaOCl

S. mutans, E. faecalis, C. albicans, and P. gingivalis.

Acid electrolyzed functional water. Acid: ACC 30 ppm; pH 2.7; ORP > 1100 mV) and alkaline: ACC 0 ppm; pH 11.5; ORP ≈ 800 mV

FW were provided by Miura Denshi (Nikaho, Japan).

NaOCl 6%.

Mean ± SD CFU/mL and Mean ± SD viable cell number.

Colony numbers of S. mutans, P. gingivalis, and E. faecalis were significantly reduced after treatment with acid FW. Alkaline FW showed strong bactericidal effects only for P. gingivalis. Further treatment for longer periods yielded a time-dependent decrease in viability; no colony was present after 20 min of treatment.

Acid FW is safe and has a bactericidal effect equivalent to that of NaOCl. Because of its efficient bactericidal, and less noxious, effects on human cells, acid FW may be a useful irrigant for effective root canal treatment.

2

Kim J., et al.; 2017 [17]

School of Dentistry, Kyungpook National University, Daegu. South Korea

1. In vitro 2. RCT

To evaluate the effect of H-water on oral Streptococci.

S. mutans and S. sobrinus

Electrolyzed Hydrogen water (H-water): dissolved hydrogen, 1.5 ppm; ORP, −600 mV ~ −700 mV.

Nanotec (nano H®, South Korea)

Tap water.

Absorbance at 590 nm.

Bacteria incubated with H-water did not form any colonies.

Oral rinse with H-water would be helpful in treating dental biofilm-dependent diseases with ease and efficiency.

3

Gunaydin M., et al.; 2014 [31]

Ondokuzmayis University, Turkey

In vitro

To investigate the in-vitro activity of superoxidized water against an extended group of microorganisms including bacteria and fungi causing hospital-acquired infections.

Six ATCC strains: A. baumannii, E. coli, E. faecalis, K. pneumoniae, P. aeruginosa, S. aureus, eight multidrug-resistant bacteria isolated from different clinical samples: A. baumannii, E. coli, vancomycin resistant E. faecium, K. pneumoniae, P. aeruginosa, methicillin-resistant S. aureus, B. subtilis, and Myroides spp.

Super-oxidized water: pH 6, 80 ppm chlorine

Medilox® (Soosan E & C, Korea) device uses salt, water and electricity and electrolyzes water.

Different concentrations and contact times of SOW.

Absence/presence of growth of ATCC strain, multidrug-resistant bacteria.

Medilox® was effective against all standard strains and all clinical isolates tested at a dilution of 1/1 and exposure time of 1 minute.

SOW produced by Medilox® disinfectant generator using water, salt, and electricity provides highly efficient disinfection.

4

Okajima M., et al. 2011 [32]

Meiji Pharmaceutical University, Tokyo, Japan

In vitro

To evaluate the bactericidal action of ERI on periodontopathic bacteria.

P.gingivalis, A. actinomycetemcomitans

Electrolyzed ion-reduced water (ERI): pH of 12.0–12.4, ORP − 344 mV, corresponding to a 0.3% NaCl solution

ERI generator (A. I. System Product Corp.)

ERI solution containing 1% Sodium carboxymethyl cellulose (CMC-Na), 0.3% NaCl solution.

CFU/mL; viable cell count of P. gingivalis and A. actinomycetemcomitans.

More than 99 and 100% of each bacteria species were killed after exposure to ERI or ERI-1% CMC-Na for 15 and 30 sec, respectively. The bactericidal action of ERI was concentration-dependent.

Results suggest that the antibacterial activity of ERI on two types of bacteria is due to the synergistic effect of a very high negative oxidation reduction potential (−344 mV) and hydroxyl radicals (OH –). This may prove extremely useful to the prevention and treatment of periodontal diseases through daily oral care, such as rinsing the mouth out with ERI and/or brushing the teeth with ERI-1% CMC-Na.

5

Jnanadev KR., et al. 2011 [33]

China Agricultural University, Beijing, Republic of China

In vitro

To investigate the efficacy of AEW and BEW in killing S. aureus imbedded in biofilm and removing established S. aureus biofilm

S. aureus

Basic electrolyzed water (BEW): pH 10.8 and 11.6, acidic electrolyzed water (AEW): pH 3.5 and 2.5

AEW and BEW were generated simultaneously from the electrolysis of a 0.1% NaCl solution in a commercial EW generator (Sai Ai Environmental Protection and Technology Development Company Ltd., Guangzhou, China).

Distilled water, NAOH and HCl, BEW generated from the electrolysis of different salt solutions (BEW1, BEW2).

% of Total number of living and dead cells (absorbance at 450 nm), % of number of living cells (absorbance at 550 nm).

As the pH of BEW rose, the removal efficacy of BEW increased. BEW at pH 10.8 reduced 42% of the biofilm mass after a 2 min treatment, whereas BEW at pH 11.6 reduced 78% of the biofilm biomass. AEW1 reduced 89% biofilm viability, whereas AEW2 only reduced 13% biofilm viability. AEW at pH 2.5 and 2% HCl solution dropped biofilm viability to 5 and 4% after a 2 min treatment, respectively.

AEW could be used as a bactericide for S. aureus imbedded in biofilm and that BEW could be applied as a removing agent for established S. aureus biofilm.

6

Gomi K., et al. 2010 [34]

Tsurumi University, Yokohama, Japan

In vitro

To evaluate the disinfection effects of functional water in comparison with sodium hypochlorite solution and hydrogen peroxide solution.

Sheep prolapsed cellulose blood as inorganic substance group and saline as organic substance group, human dental pulp cells

Alkaline (pH 12.3), Strong acid (pH 2.8; 10 ppm Chlorine) and Hypochlorous (pH 6.0; 50 ppm Chlorine) electrolysis water (AEW, SAEW, or HAW)

AEW (Aoi Engineering), SAEW (Aoi Engineering), or HAW (Technomak)

Physiologic saline (PS), 3% sodium hypochlorite solution (SHS), 3% hydrogen peroxide solution (HPS).

The number of living cells (absorbance at 490 nm). CFU/mL of E. faecalis.

SAEW showed a weaker microbicidal activity compared to HAW in the absence of organic substance, and complete inhibition of colony-forming bacteria did not occur in the presence of organic substance. The microbicidal effect was not observed with AEW and PS.

Functional waters SAEW and HAW have good microbicidal effect in the presence of organic substance, with disinfection activity similar to that of SHS.

7

Yamada K., et al. 2010 [35]

Tokyo Dental College, 1–2-2 Masago Mihama-ku, Chiba, Japan

In vitro

To investigate the efficacy of super-oxidised water containing a high concentration of O (O-water) in destroying cariogenic and periodontopathic bacteria.

S. sobrinus, P. gingivalis, P. intermedia, A. actinomycetemcomitans, and F. nucleatum.

Super-oxidised water at low, medium or high concentration

O-water was generated in the AOE-750 (Oxy Japan Corporation, Tokyo, Japan). Three concentrations of O⁻water—low (pH 3.0), medium (pH 2.5, O⁻ concentration: ca. 3.2 x low) and high (pH 2.0, O⁻ concentration: 10x low)—were prepared.

Distilled water, hydrochloric acid solution.

CFU/mL of cariogenic and periodontophatic bacteria.

Super-oxidised water showed bactericidal activity against all cariogenic and periodontopathic bacteria tested in this study. The antibacterial effect at 37° C was higher than that at room temperature.

O-water exerts an antibacterial effect on cariogenic and periodontopathic bacteria, suggesting its potential as a disinfectant in the prevention of bacterial contamination of dental equipment

8

Ileri C., et al. 2006 [translated from Turkish] [36]

High Technology Institute, Gebze, Turkey

In vitro

To investigate the effects of EAAW on standard strains of pathogenic microorganisms at different time periods and at different concentrations.

S. aureus and P. aeruginosa

Electro-activated acidic water (EAAW) at different concentrations

EAAW was produced by direct current passed through the water by using a power source.

Sterile deionized water.

Log CFU/mL of S.aureus and P. aeruginosa.

A decrease of approximately 6.5–8.2 Log CFU/mL was detected at 2-100% concentrations of EAAW at the 10th second, depending on the microorganisms.

EAAW can be used in surface disinfection even at low (at least 2%) dilutions.

9

Gulabivala K. 2004 [22]

Seoul National University, Seoul, Korea

In vitro

To evaluate the antibacterial effect of electrolyzed tap water (Puriwater) on five major periodontopathogens cultured in vitro.

A. actinomycetemcomitans, F. nucleatum, P. gingivalis, P. intermedia, and T. denticola.

Puri-water: pH 8.4.

Tap water was subjected to electrolysis (30 V of DC/300 mA) for 2 min at ambient temperature using an electrolysis apparatus equipped with platinum electrodes (SciacuaTM, Puri Co., Korea).

Tap water.

CFU/mL or OD₆₆₀

Puri-water reduced the bacterial counts to 12.6–15.4% for A. actinomycetemcomitans, F. nucleatum, P. intermedia, and P. gingivalis. Growth of T. denticola was not observed during 7-day incubation after exposure.

Electrolyzed tap water markedly inhibited the growth of cultured periodontopathogens.

10

Vorobjeva NV., et al. 2004 [37]

Lomonosov Moscow State University

In vitro

To evaluate the bactericidal effect of EO water on common hospital bacterial strains in vitro.

P.aeruginosa, E. faecalis, S. aureus, E. coli, Bacillus cereus (vegetative cells and spores), Citrobacter freundii, Flavobacter sp., Proteus vulgaris, Alcaligenes faecalis, and Aeromonas liquefaciens.

Electrolyzed Oxidizing Water (EO):.84 ± 0.01, 1125 ± 3 mV, 43 ± 0.3 ppm Chlorine, and 4.0 ± 0.02 mS/cm

EO water was obtained from ROX-20TA electrolyzer, Hoshizaki Electric Company (Aichi, Japan) at 19.8 A and 10 V.

Deionized water.

Mean Log CFU/ mL.

The counts of the majority of the bacterial strains in the treatment samples were reduced to zero after 0.5 min of treatment, whereas the population of B. cereus was 3.76 log CFU/mL. After 5 min,the counts of the vegetative cells and spores of B. cereus were zero.

EO water renders a strong bactericidal action to both Gram-positive and Gram-negative bacteria as well as to the vegetative cells and spores of bacilli.

11

Shimada K., et al. 2000 [38]

Nihon University Japan

In vitro

To compare the bactericidal effects on cariogenic and periodontopathogenic bacteria of EW.

S. mutans, S. sobrinus, S. mitis, S. salivarius, S. sanguis, A. biscosus, A. naeslundii, F. nucleatum, P. gingivalis, P. nigrescens, P. loeschii, P.melaninogenica, and A. actinomycetemcomitans.

Acid oxidizing water (AOW), neutral oxidizing water (NtOW), acid oxidizing water with a low chlorine ppm

AOW was prepared using an Aquachid NDX- 60KMW electrolysis apparatus, (Omco O.M.C. Co., Saitama, Japan), NtOW using an Aquachid NDX- 60KH (Omco O.M.C. Co.), and AOW-LC using a Minestar 201 (Minestar Co., Tokyo, Japan).

Povidoneiodine (PI) 0.35, 0.2% chlorhexidine (CHX), Listerine (LST), 70% ethyl alcohol (Et), PBS.

CFU/mL

Bacteria incubated with NtOW, AOW, AOW-LC, LST, PI or Et did not form any colonies.

The three types of oxidizing water examined are approximately as potent at inhibiting bacterial plaque formation as conventional chemical plaque control agents.

12

Horiba N., et al. 1999 [39]

AICHI-GAKUIN University, Nagoya, Japan

In vitro

To examine Electrolized Neutral Water (ENW) bactericidial effect against bacteria isolate from infected root canals.

Methicillin-resistant S.epidermidis, isolated from a human nasal cavity; B. subtilis. 15 strains isolated and identified from infected root canals: S. aureus, S. sanguis, L. acidophilus, S. intermedia, P. niger, P.anaerobius, V. parvula, L. rogosae, A. israelii, E. lentum, B. bifidum, P. acnes, P. endodontalis, P. melaninogenica, and F. nucleatum.

Electrolized Neutral Water (ENW)

Ameni Clean (National Co/Matsushita Seico Co, Osaka) was used to produce ENW.

Sterilized distilled water.

CFU/mL

ENW was bactericidal against 12 strains: S aureus, MRSE, S sanguis, P niger, Ps anaerobius, L acidophilus, L. rogosae, A israelii, E lentum, B bifidum, P. endodontalis, and F. nucleatum. In addition, the ENW reduced the bacterial numbers of the other 6 strains. However, ENW showed little effectiveness against B subtilis.

ENW exhibits a bacteriostatic/bactericidal action against isolates obtained from infected root canals

13

Tanaka H., et al. 1996 [40]

Nagasaki University School of Medicine, Nagasaki, Japan

In vitro

To evaluate the antimicrobial activity of superoxidized water against Gram-positive and Gram-negative bacteria.

Methicillin-sensitive S.aureus, methicillin-resistant S.aureus, S. epidermidis, S. marcescens, E. coli, P. aeruginosa, and B. cepacia.

Superoxidized water: pH 2.3–2.7, ORP 1000-l 100 mV, 30 ppm chlorine.

Superoxidized water was prepared by electrolysis of tap water using the Super Oxseed alpha 1000 (Janix, Inc., Kanagawa, Japan).

0.1% chlorhexidine, 0.02% povidone iodine, 80% ethanol and sterile distilled water.

CFU/ mL

Superoxidized water reduced the viable count below the limit of detection within 10 s of contact as did 80% ethanol and 0.02% povidone iodine. Superoxidized water killed B. cepacia faster than 0.02% povidone iodine. The bactericidal activity of superoxidized water was superior to that of 0.1% chlorhexidine and 0.02% povidone iodine.

Superoxidized water has powerful bactericidal activity and is a low cost but powerful disinfectant.

14

Ogawa T., et al. 1998 [translated from Japanese] [42]

The Nippon Dental University, Japan

In vitro

To evaluate the bactericidal effect of soft alkaline solution water on periodontopathic bacteria.

A. actinomycetemcomitans, P. gingivalis, P. intermedia, and E.coli

Soft alkaline solution water: ORP 800 mV, pH 8–06, 60 ppm Chlorine

OXILIZER generator: OXM01 (Miura Electronics Corporation, Tokio)

Hard oxidized water: 1050 mV, pH 2.46, Chlorine concentration 30 ppm.

CFU/mL after contact time (1,5, and 10 min).

Soft alcaline water and strong acid water had a bactericidal effect against all bacteria, after 5 and 10 minutes (p < 0,05), and beyond 10 minutes (p < 0,01).

Soft alcaline water is as effective as strong acid water, having no problem with the presence of saliva. Strong acid water could be more effective in the oral environment.

15

Ogiwara K., et al. 1996 [translated from Japanese] [41]

Nippon Dental University, Japan

In vitro

To evaluate the bactericidal effect of AAW (Aqua alkalic water) to periodontopatic bacteria.

S. aureus, E. coli, A. actinomycetemcomitans, P. gingivalis, P. intermedia, and F. nucleatum

Aqua alkalic water (AAW), Aqua oxidized water (AOW),

OXILIZER water generator (OXILIZER Co., Tokyo, Japan).

Saline solution.

CFU/mL

Aa, Pg, Pi were killed in one minute (Aa; 108 to 104 CFU/ml, Pg; 108 to 103 CFU/ml, Pi; less than 108 to 103 CFU/ml) from original 100% AAW, while it took more than one minute to kill Fn, and Sa and Ec which survived after ten minutes.

The disinfectant effect of AAW could be useful for the treatment of periodontal diseases.

16

Hsieh SC., et al. 2020 [43]

Taipei Medical University, Taiwan

In vitro

To investigate the antibacterial property and cytotoxicity of EO water containing HOCl, relative to NaOCl.

Zebrafish embryo, S. mutans and E. faecalis

Electrolyzed Oxidizing (EO), ORP 1100 mV, 330–350 Chlorine

The ANK-Neutral Anolyte GH-40 (Envirolyte Industries International Ltd., Tallinn, Estonia) was used to produce EO water by mixing water with an over-saturated solution of sodium chloride under 110 V.

1.5% Sodium Hypochlorite, E3 medium.

CFU/mL

All the HOCl or NaOCl treatment groups showed over a 5 log 10 cfu/mL reduction in E. faecalis and S. mutans population, indicating > 99.9% antibacterial efficacy.

Both EO waters containing 0.0125 and 0.0250% HOCl revealed a remarkable but similar bactericidal effect (>  99.9%) to that of conventional NaOCl against E. faecalis and S. mutans.

17

Salisbury AM., Percival SL. 2019 [44]

5D Health Protection Group Ltd., Centre of Excellence for Biofilm Science (CEBS), Liverpool, UK

In vitro

To assess the antimicrobial and anti-biofilm efficacy of a new formulation of electrolysed water against microorganisms associated with complicated chronic wounds.

CDC Biofilm Bioreactor Model S. aureus and P. aeruginosa

Electrolysed water at concentrations of 100, 75, 50 and 25%

Water was produced by passing an electric current through deionised water, with the 2 main constituents being Sodium hypochlorite and Hypochlorous acid.

0.85% sodium chloride solution.

Mean Log₁₀ densitiy CFU/mL.

No colonies of either S. aureus or P. aeruginosa were detected following antimicrobial treatment with all dilutions of the electrolysed water (p < 0.0001).

The assessment of the electrolysed water as an antimicrobial and antibiofilm agent showed exceptionally fast-acting efficacy. The use of electrolysed water as a method of controlling bioburden and biofilm in complicated chronic wounds could significantly aid wound closure.

18

Kim, S.B. 2016 [45]

Korea Institute of Industrial Technology, Ansan, Republic of Korea

In vitro

To propose a low-level hypochlorous acid solution of electrolyzed water as an alternative to mouthwash by monitoring oral bacteria though a bactericidal activity experiment

P. gingivalls, P. intermedius, P. nigrescens, F. nucleatum, S. mutans, S. sobrinus, S. godonii, S. oralis, S. salivarius

Low-level HOCL acid solution (electrolyzed water): pH 5–7, 3–5 mg/L Chlorine

Water was produced through a macroporous structure at Pt films electrode and an electrolysis device.

Non-treated saline solution served.

Bactericidal activity (%) for 1 min.

The low-level hypochlorous acid solution exhibited ≥99.9% bactericidal activity for all strains tested.

Low-level hypochlorous acid solution for the range of bacteria tested exhibits greater bactericidal activity for four anaerobic bacteria responsible for periodontitis and five facultative anaerobic bacteria associated with cavity development.

19

Davis JM., et al. 2007 [46]

Marquette University, Milwaukee, Wisconsin, USA

In vitro

To compare the antimicrobial action of Dermacyn, BioPure MTAD, 2% CHX (Ultradent, West Jordan, UT), and 5.25% NaOCl against E. faecalis.

E. faecalis

Super-oxidized water

Dermacyn (Oculus Innovative Sciences, Petaluma, CA, USA).

BioPure MTAD, 2% CHX, and 5.25% NaOCl, sterile distilled water.

Mean zones of microbial inhibition on aerobical and aerobical conditions.

Dermacyn and the control showed zones of microbial inhibition that were not different from each other (p > 0.05).

Dermacyn showed no ability to prevent the growth of E. faecalis.

20

Landa-Solis C., et al. 2005 [47]

Instituto Nacional de Rehabilitación, Secretarıía de Salud, Mexico, DF, Mexico

In vitro

To evaluate the disinfectant activity of Microcyn against various microbes including pathogenic vegetative bacteria in vitro.

E. coli, S. aureus, P. aeruginosa, S. typhi,

Super-oxidized waters (SOWs)

Microcyn (SOW) is made up by purified water which passes through anode and cathode chambers that are separated from a middle salt (NaCl) chamber by ionic membranes in a REDOX equipment (Oculus Innovative Sciences, California, USA).

Sterile, deionized water.

Log₁₀ surviving Bacillus spores.

An exposure time of 30 s was enough to completely inactivate all pathogens tested in thetreatment samples (99.9999% reduction). Thus, a log10 reduction factor of 8 in the level of all pathogens occurred in the treatment samples.

Microcyn is an effective disinfectant for which sporocide activity and appropriate applications are now being validated.

21

Yoo, Y.S., et al. 2015 [49]

Dankook University, Cheonan, Republic of Korea

In vitro

To analyze and compare the antimicrobial activity of electrolyzed water using various electrodes on biofilms of oral microbes

Biofilms of oral microbes (pooled saliva of 10 healthy donors) and planktonic oral microbes. (To form biofilm of P. gingivalis and T. forsythia, the protein of F. nucleatum was extracted)

Electrolyzed Water using copper (EWC), silver (EWS) and platinum (EWP) electrode

Tap water was subjected to electrolysis for 5 min with 24 V of DC 400 mA using copper, silver or platinum electrode in an whole tank undivided anode chamber and cathode chamber.

Tap water.

CFU/mL, OD 590 nm and live/dead staining of S. mutans, F. nucleatum, P. gingivalis, T. forsythia.

Electrolyzed water using platinum electrode (EWP) exhibited antimicrobial activity against S. mutans, P. gingivalis and T. forsythia. The electrolyzed water using copper electrode (EWC) and silver electrode (EWS) did not affect oral microbes. EWP showed strong antimicrobial activity against the biofilm of oral microbes.

Electrolyzed water generated using a palladium electrode may have potential value as a gargle solution for prevention of oral diseases induce by pathogens and denture-related stomatitis.

22

Cho I.W., et al. 2017 [50]

College of Dentistry, Dankook University, Korea

In vitro

To investigate antimicrobial activity of recent developed EW generator for oral bacteria

A. actinomycetemcomitans, S. mutans, F. nucleatum, and P. gingivalis.

Hydrogen enriched electrolyzed water

Tap water was subjected to electrolysis for 3 min with 24 V of DC using eBio-Cleaner (ebiotech, Seoul) .

Tap water and Listerine®.

CFU/mL, LIVE/DEAD staining of cariogenic and periodontopathogenic bacteria.

eBio cleaner water showed significantly antimicrobial activity against S. mutans compared to tap water. The levels of F. nucleatum, P. gingivalis, and A.a were reduced by eBio-cleaner water. eBio-cleaner water reduced the levels of P. gingivalis and A.a by several hundred-fold compared to tap water.

The electrolyzed water generated by eBio cleaner reduced the growth of periodontopathogens and S. mutans. The EW generated by eBio-cleaner showed disruptive and antimicrobial effect on the salivary biofilm.

23

Lee K. 2016 [51]

Dankook University, Korea

In vitro

To investigate the antimicrobial effects of NEW on cariogenic bacteria and their biofilm, and to compare the antimicrobial activity of NEW and commercial gargle solution

S. mutans and S. sobrinus (To form biofilm, saliva was used).

Two types of Neutral Electrolyzed Water containing 0.05 and 0.15% sodium chloride

EW was generated from an electrolyzing distilled water containing 0.05 and 0.15% sodium chloride in an undivided anode chamber and cathode chamber.

Distilled water and commercial gargle solutions (alcohol-containing gargle for adults and a fluoride-containing gargle for children).

CFU and OD 590 nm of cariogenic bacteria.

The EW showed significant antimicrobial activity against S. mutans and S. sobrinus. Furthermore, the EW has more antimicrobial activity compared to the gargle solution for children. The EW and the alcohol-containing gargle significantly disrupted the biofilm. Furthermore, the count of S. mutans and S. sobrinus in the biofilm was decreased by both the EW and the alcohol-containing gargle. Interestingly, the EW disrupted more the biofilm and killed more the cariogenic bacteria in the biofilm than the alcohol-containing gargle.

The NEW is effective in removing cariogenic biofilm as thoroughly as the commercial gargle solutions and showed antimicrobial activity against S. mutans and S. sobrinus.

24

Gupta M., et al. 2017 [52]

Department of Microbiology, IMS, BHU, Varanasi, U.P., India

In vitro

To observe the effect of SOW in different dilutions against several pathogenic bacteria

S. aureus, E. coli, P. aeruginosa, A. baumannii, E. faecalis, and K. pneumoniae.

Superoxidised water (SOW), pH 5.0–6.5, ORP > 950 mv

SOW (Sterisol) generated by Steri-Gen® disinfectant generating system, prepared by passing the normal saline over titanium coated electrode at 9 amp

5 and 10 times dilutions of SOW.

Growth inhibition or no growth inhibition.

Undiluted SOW and 5, 10 times dilution of SOW inhibited the growth of bacteria.

Undiluted SOW prevent the occurrence of nosocomial infections. Also, the efficacy of SOW was observed in different dilutions against various microbes.

25

Lucio-Sauceda, D.G., et al. 2019 [53]

Autonomous University of Nuevo Leon, México.

In vitro

To evaluate the antimicrobial and antibiofilm activity of a novel ESS against H. pylori.

H. pylori (ATCC 700,824)

Electrolyzed solutions of superoxidation (ESS): ph 6–7, 15 ppm Chlorine, ORP 650–900 mV with

OxOral®^, was elaborated and supplied by Esteripharma (Mexico city, Mexico).

Tetracycline, saline solution.

Mean ± (SD) of MIC (%), The minimum bactericidal concentration (MBC value, % of biofilm inhibition).

An important antimicrobial activity of OxOral® was observed from 15 to 3.75 ppm, in those doses no difference was observed compared to 5 μg/mL tetracycline (p > 0.05). Therefore, 3.75 ppm of OxOral® with 99.7 ± 7.7% inhibition was established as the MIC value. The MBC value was 7.5 ppm when no visible growth was observed. However, when biofilm cells were treated with 0.938 ppm, 0.469 ppm, and 0.234 ppm of OxOral® values of 99.9 ± 5.5%, 89.1 ± 1.2%, and 35.5 ± 0.9% and of inhibition were obtained, respectively.

Antimicrobial and anti-biofilm effect of OxOral® mouthwash against H. pylori, and low cytotoxicity open the possibility of its therapeutic use in H. pylori-infected patients as adjuvant in conventional therapy.

26

Park YK., et al.; 2013 [translated from Korean] [54]

Gimcheon University, Gimcheon, Korea

In vitro

To evaluate the antibacterial effect of slightly acidic electrolyzed water (SAEW) for use as a mouth-rinse on seven oral pathogens.

S. mutans, S. sobrinus, A. actinomycetemcomitans, P. gingivalis, P. intermedia, F. nucleatum, and E.coli

Slightly acidic electrolyzed water (SAEW): pH 6.2, ORP 728–800 mV, 30 mg/L ppm chlorine

SAEW was produced in a device with hipochlorous acid (BC-360, Cosmic Round Korea Co., Seongnam, Corea)

Listerine, clorhexidine, tap water.

Mean/SD:Diameter of clear zone (mm), MIC (mg/ml or ml/ml), MBC (mg/ml or ml/ml), Bacterial strains (CFU/ml).

SAEW showed potent antimicrobial activity with a MIC value of 0.0075–0.015 mg/mL and a MBC value of 0.015–0.03 mg/mL. Mouth rinsing with SAEW showed 99.9% bacterial inhibition.

SAEW exhibited potent antimicrobial activity against all oral pathogens causing dental caries and periodontal disease.

27

Yanik K., et al. 2015 [55]

Ondokuz Mayis University, Faculty of Medicine, Kurupelit, Samsun, Turkey

In vitro

To reveal the in-vitro effect of electrolyzed water against nosocomial bacteria under different concentrations.

A. baumannii, E. coli, E. faecalis, K. pneumoniae, P. aeruginosa, S. aureus and eight different multidrug resistant bacteria: A. baumannii, E. coli, vancomycin resistant Enterococcus faecium, K. pneumoniae, P. aeruginosa, methicillin resistant S. aureus, Bacillus subtilis, and Myroides spp.

Electrolyzed water, pH 6.5 to 7.5, 50–700 ppm chlorine

Electrolyzed water produced by the Envirolyte® (Envirolyte® Industries International Ltd., Estonia).

Different concentrations of EW (1/1, 1/2, 1/10, 1/20, 1/50, 1/100).

No growth(−) or bacteria growth(+).

It was observed that the 1/1, 1/2, and 1/10 dilutions of electrolyzed water were effective on the standrard strains for all intervals of time.

Super-oxidized water is effective on a broad spectrum of bacteria. It was observed that Envirolyte® electrolyzed water of 100% concentration would be convenient to use for disinfection when diluted to a concentration of 1/10.

28

Vahabi S., et al. 2020 [56]

Shahid Beheshti University of Medical Sciences, Tehran, Iran

In vitro

To compare the antimicrobial effects of EW on the microorganisms of microbial plaque.

S. salivarius, S. aureus, L. casei, and A. actinomycetemcomitans (JP2).

EW was prepared at three pH values, pH 3, MBEW mildly basic electrolyzed water pH 9, and BEW-pH 11.

EW was produced using 7-plate water ionizer (iWater-sharp, Korea)

PBS,Chlorhexidine (CHX) 0.2%, CHX 0.009%, distilled water.

Mean value of CFUs and % reduction.

All EW types had a significant (P < 0.001) and strong antibacterial efficiency against all species in the AEW, MBEW, and BEW groups compared with the negative control. The maximum antibacterial activity of the EW at pH 3, pH 9, and pH 11 among the selected bacterial species was against A.a with a bacterial reduction of 100, 99.3, and 100%, respectively. The least antibacterial activity at those pH values was against S. aureus with a bacterial reduction of 98.04, 89.16%, and 88.75, respectively. At three EW types had an equal antibacterial potency against L. casei with 99.99% CFU/mL reduction. The CFU reduction of S. salivarius was 99.92, 99.3, and 99.94% for EW types with AEW, MBEW, and BEW, respectively.

Efficient antibacterial activities of EW at different pH values against four oral bacterial species within a 2-hour time span between water preparation and the test procedure were detected.

29

Goishi T., et al. 1996 [translated from japanese] [58]

Department of Preventive and Community Dentistry, School of Dentistry at Tokyo, The Nippon Dental University

In vitro

To investigate the bactericidal effects of AOW.

S. mutans and E. coli

Aqua Oxidizing Water (AOW), undiluted and diluted (20, 30, 40, 50%)

Water and salt was added to an electrolysis device (OXILYZER90XM-01Ltd.).

Tap water, PBS.

Number of viable bacteria CFU/mL.

Undiluted AOW was effective against the two strains tested, and the same effect was observed with 20% AOW (5-fold dilution) treatment for 30 sec.

These results suggest that 20% AOW which has bactericidal activity and is not toxic to gingival cells may be useful for dental clinical application.