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Chromographic Analysis and Cytotoxic Effects of Chlorhexidine and Sodium Hypochlorite Reaction Mixtures

Nocca G, Ahmed HMA, Martorana GE, Callà C, Gambarini G, Rengo S, et al.

J Endod, 2017 Sep;43(9):1545-1552.
PMID: 28734651 DOI: 10.1016/j.joen.2017.04.025

INTRODUCTION: The literature reveals controversies regarding the formation of para-chloroaniline (PCA) when chlorhexidine (CHX) is mixed with sodium hypochlorite (NaOCl). This study aimed to investigate the stability of PCA in the presence of NaOCl and to examine the in vitro cytotoxic effects of CHX/NaOCl reaction mixtures.
METHODS: Different volumes of NaOCl were added to CHX (mix 1) or PCA (mix 2). Upon centrifugation, the supernatant and precipitate fractions collected from samples were analyzed using high-performance liquid chromatography. The cytotoxic effects of both fractions were examined on human periodontal ligament and 3T3 fibroblast cell lines.
RESULTS: High-performance liquid chromatographic analysis showed no PCA signal when NaOCl was mixed with CHX (mix 1). In mix 2, the intensity of PCA was decreased when NaOCl was added to PCA, and chromatographic signals, similar to that of CHX/NaOCl, were also observed. The mortality of precipitates exerted on both cell lines was lower compared with that of supernatants.
CONCLUSIONS: The discrepancy in the data from the literature could be caused by the instability of the PCA in the presence of NaOCl. The CHX/NaOCl reaction mixture exhibits a wide range of cytotoxic effects.

Matched MeSH terms: Sodium Hypochlorite/pharmacology*
The Efficacy of Apple Cider Vinegar at Different pH Values as an Antimicrobial Agent: An In Vitro Study

Chandraseharan P, Sockalingam SNM, Shafiei Z, Zakaria ASI, Mahyuddin A, Rahman MA

J Contemp Dent Pract, 2023 Oct 01;24(10):779-786.
PMID: 38152911 DOI: 10.5005/jp-journals-10024-3581

AIMS AND BACKGROUND: This study evaluates the antimicrobial activities of commercially available 5% apple cider vinegar (ACV) against Enterococcus faecalis, Streptococcus mutans, and Lactobacillus casei. Materials and methods: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were conducted using the broth microdilution method. Sodium hypochlorite (NaOCl) of 5.25% was used as a positive control, and comparisons were also made with acetic acid (AA) as the main ingredient in ACV. The three test bacteria treated with the most effective ACV dilution were visualized under a transmission electron microscope (TEM) for structural changes.
RESULTS: Minimal inhibitory concentration was determined at 0.625% of the concentration of ACV against S. mutans and E. faecalis and 1.25% of the concentration of ACV against L. casei with two-fold serial dilutions. A concentration of 5 × 10-1% with 10-fold serial dilutions was found to be the MIC value for all three bacteria. No significant differences were found when compared with the positive control (NaOCl) (p = 0.182, p = 0.171, and p = 0.234), respectively, for two-fold serial dilutions and (p = 1.000, p = 0.658, and p = 0.110), respectively for 10-fold serial dilutions. MBC was observed to be 5% ACV for both E. faecalis and S. mutans. However, positive microbial growth was observed on the agar plate when cultured with L. casei. An independent sample t-test showed no significant differences (p > 0.05) in the antimicrobial activities between 5% ACV and 5% pure AA. TEM revealed cell wall and cytoplasmic membrane disruptions on all three bacteria at MIC value.
CONCLUSION: Apple cider vinegar has antimicrobial activities against Enterococcus faecalis, Streptococcus mutans, and Lactobacillus casei at their respective MIC values.
CLINICAL SIGNIFICANCE: Apple cider vinegar can be an alternative antimicrobial dental pulp disinfectant to sodium hypochlorite. Apple cider vinegar can be used safely, especially in children's dental pulp therapy and deep caries management, when adequate tooth isolation is not readily achievable. Thus, adverse reactions commonly associated with other frequently used chemical disinfectants can be avoided.

Matched MeSH terms: Sodium Hypochlorite/pharmacology
Fulltext Antimicrobial Efficacy of Fruit Peels Eco-Enzyme against Enterococcus Faecalis: An In Vitro Study

Mavani HAK, Tew IM, Wong L, Yew HZ, Mahyuddin A, Ahmad Ghazali R, et al.

Int J Environ Res Public Health, 2020 Jul 15;17(14).
PMID: 32679828 DOI: 10.3390/ijerph17145107

Sodium hypochlorite (NaOCl), an effective endodontic irrigant against Enterococcus faecalis (EF), is harmful to periapical tissues. Natural pineapple-orange eco-enzymes (M-EE) and papaya eco-enzyme (P-EE) could be potential alternatives. This study aimed to assess the antimicrobial efficacy of M-EE and P-EE at different concentrations and fermentation periods against EF, compared to 2.5% NaOCl. Fermented M-EE and P-EE (3 and 6 months) at various concentrations were mixed with EF in a 96-well plate incubated for 24 h anaerobically. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of M-EE and P-EE were determined via EF growth observation. EF inhibition was quantitatively measured and compared between different irrigants using the one-way analysis of variance (ANOVA), and different fermentation periods using the independent-samples T-test. M-EE and P-EE showed MIC at 50% and MBC at 100% concentrations. There was no significant difference in antimicrobial effect when comparing M-EE and P-EE at 50% and 100% to 2.5% NaOCl. P-EE at 6 months fermentation exhibited higher EF inhibition compared to 3 months at concentrations of 25% (p = 0.017) and 0.78% (p = 0.009). The antimicrobial properties of M-EE and P-EE, at both 100% and 50% concentrations, are comparable to 2.5% NaOCl. They could therefore be potential alternative endodontic irrigants, but further studies are required.

Matched MeSH terms: Sodium Hypochlorite/pharmacology
Effectiveness of ultrasonic files in the disruption of root canal bacteria

Ahmad M, Pitt Ford TR, Crum LA, Wilson RF

Oral Surg. Oral Med. Oral Pathol., 1990 Sep;70(3):328-32.
PMID: 2216361

The physical mechanisms of ultrasound, namely cavitation and acoustic streaming, generated by the Enac-Osada ultrasonic unit were investigated for effectiveness in disrupting Streptococcus mitis. In addition, the bactericidal effect of ultrasound in the presence of 2.5% sodium hypochlorite was examined. Bacterial suspensions were irradiated directly with ultrasound in simulated root canals, and the viability of bacteria was examined after growth on a blood agar medium under anaerobic conditions at 37 degrees C for 5 days. The results indicated that ultrasound per se failed to disrupt bacteria but resulted in increases in the viable counts; the former was considered to be because of the lack of cavitation and the latter because of the dispersal effects of acoustic streaming. The 2.5% sodium hypochlorite solution demonstrated powerful bactericidal activity.

Matched MeSH terms: Sodium Hypochlorite/pharmacology
Trisodium phosphate and sodium hypochlorite are more effective as antimicrobials against Campylobacter and Salmonella on duck as compared to chicken meat

Sarjit A, Dykes GA

Int J Food Microbiol, 2015 Jun 16;203:63-9.
PMID: 25791251 DOI: 10.1016/j.ijfoodmicro.2015.02.026

Little work has been reported on the use of commercial antimicrobials against foodborne pathogens on duck meat. We investigated the effectiveness of trisodium phosphate (TSP) and sodium hypochlorite (SH) as antimicrobial treatments against Campylobacter and Salmonella on duck meat under simulated commercial water chilling conditions. The results were compared to the same treatments on well-studied chicken meat. A six strain Campylobacter or Salmonella cocktail was inoculated (5 ml) at two dilution levels (10(4) and 10(8) cfu/ml) onto 25 g duck or chicken meat with skin and allowed to attach for 10 min. The meat was exposed to three concentrations of pH adjusted TSP (8, 10 and 12% (w/v), pH 11.5) or SH (40, 50 and 60 ppm, pH 5.5) in 30 ml water under simulated spin chiller conditions (4 °C, agitation) for 10 min. In a parallel experiment the meat was placed in the antimicrobial treatments before inoculation and bacterial cocktails were added to the meat after the antimicrobial solution was removed while all other parameters were maintained. Untreated controls and controls using water were included in all experiments. Bacterial numbers were determined on Campylobacter blood-free selective agar and Mueller Hinton agar or xylose deoxycholate agar and tryptone soya agar using the thin agar layer method for Campylobacter and Salmonella, respectively. All TSP concentrations significantly (p<0.05) reduced numbers of Campylobacter (~1.2-6.4 log cfu/cm(2)) and Salmonella (~0.4-6.6 log cfu/cm(2)) on both duck and chicken meat. On duck meat, numbers of Campylobacter were less than the limit of detection at higher concentrations of TSP and numbers of Salmonella were less than the limit of detection at all concentrations of TSP except one. On chicken meat, numbers of Campylobacter and Salmonella were less than the limit of detection only at the lower inoculum level and higher TSP concentrations. By contrast only some of the concentrations of SH significantly (p<0.05) reduced numbers of Campylobacter and Salmonella (~0.2-1.5 log cfu/cm(2)) on both duck and chicken meats. None of the SH treatments resulted in numbers of either pathogen being less than limit of detection. Results indicate that chicken meat has the ability to effectively protect Campylobacter and Salmonella against the impact of trisodium phosphate and sodium hypochlorite while duck meat does not. This study suggests that trisodium phosphate has a strong potential for application in a commercial poultry processing to reduce Campylobacter and Salmonella specifically on duck meat.

Matched MeSH terms: Sodium Hypochlorite/pharmacology*