Introduction: Oral healthcare professionals play a crucial role in guiding patients toward evidence-based choices among the many available oral rinses. In this study, we explored how specific oral rinse formulations affect the viability and modulate critical virulence traits of the opportunistic fungal pathogen Candida albicans.Materials and Methods: We assessed the effects of these oral rinses on the production of germ tube, production of phospholipase and hemolysin, as well as biofilm formation. Results: We found that oral rinses containing cetylpyridinium chloride (CPC) and chlorhexidine (CHX) showed the greatest fungicidal activity with the lowest minimum fungicidal concentrations (0.38% and 0.78%, respectively). Oral rinses based on zinc chloride and sodium fluoride with Miswak bark extract (MIS) or essential oils (EO) had much lower fungicidal activity (8–16 times lower) compared to CHX and CPC. However, they had a significantly greater impact on the virulence traits of C. albicans. They reduced germ tube production by 86–89% (vs. 42% for CHX and 29% for CPC), completely inhibited phospholipase and hemolysin production, and together with the CPC-based oral rinse, exerted the greatest reductions in biofilm formation across all tested concentrations. This was in contrast to both the controls and CHX, which had a minimal effect on biofilm formation. Conclusion: By inhibiting the virulence factors, the oral rinse can have a crippling effect on C. albicans, weakening this opportunistic pathogen and hindering its potential to cause infection.

Highlights of the Study

  • Oral rinses with cetylpyridinium chloride and chlorhexidine showed the most potent fungicidal activity.

  • Zinc chloride and sodium fluoride-based rinses with Miswak bark extract or essential oils significantly affected the virulence traits of C. albicans.

  • Chlorhexidine-based oral rinse had only a minimal effect on biofilm formation, in contrast to the other formulations which exerted high anti-biofilm activity.

As a commensal yeast, Candida albicans (C. albicans) is part of the microbiota of the human body. However, in times of dysbiosis, it reverts to an opportunistic pathogen causing mucosal or systemic disease, which is especially life-threatening in vulnerable populations, with a high rate of mortality even with appropriate antifungal treatment [1‒4]. Various factors increase the risk of oral Candida infections, including broad-spectrum antimicrobial therapy, chemotherapy, diabetes, and HIV-induced immunosuppression [2]. Additionally, patients with dentures, orthodontic devices, or issues like hyposalivation and persistent mouth dryness face elevated risks [5]. Candida survival in the host is aided by factors like adherence to host tissues and prosthetic devices, morphological switching [6], and the release of hydrolytic enzymes such as phospholipase and hemolysin [7‒9], along with biofilm formation [10]. These mechanisms enhance the ability of Candida to persist in the host and resist both antifungal agents and immune defenses, increasing the likelihood of systemic infection.

Despite the widespread use of oral rinses aimed at reducing microbial load [11‒13], their impact on C. albicans virulence factors remains poorly understood. While previous studies have addressed their influence on growth [13], their effects on virulence factors remain unclear. This study aims to fill this knowledge gap by assessing the effects of oral rinses with different formulations on the growth and virulence factors of C. albicans. The investigation will focus on germ tube and biofilm formation, as well as phospholipase and hemolysin production. By concentrating on a single ATCC Candida strain, we aimed to provide precise insights into the effects of specific oral rinses on a well-defined strain. Through this approach, our study aimed to contribute to a deeper understanding of oral candidiasis and potentially inform future clinical applications in oral health.

C. albicans and Oral Hygiene Products

C. albicans (ATCC 10231) was routinely cultivated on sabouraud dextrose agar and broth (SDA; SDB) at 37°C for 24 h. The oral rinses listed in “Table 1” were purchased from the local market. All procedures were repeated three times in triplicate for consistency.

Table 1.

Commercial names and active ingredients present in the oral rinses

Commercial names (abbreviations)Active components
Gar Garol (CHX) Chlorhexidine gluconate (0.2%), methyl salicylate, chlorbutol, ethanol 
Colgate Plax (CPC) Sodium fluoride (225 ppmF), cetylpyridinium chloride (0.075%), potassium sorbate, menthol 
Listerine Miswak (MIS) Zinc chloride, Salvadora persica bark extract, sodium fluoride (220 ppmF) 
Listerine Total care (EO) Eucalyptol, zinc chloride, benzoic acid, sodium benzoate, methyl salicylate, thymol, sodium fluoride (220 ppmF), menthol 
Commercial names (abbreviations)Active components
Gar Garol (CHX) Chlorhexidine gluconate (0.2%), methyl salicylate, chlorbutol, ethanol 
Colgate Plax (CPC) Sodium fluoride (225 ppmF), cetylpyridinium chloride (0.075%), potassium sorbate, menthol 
Listerine Miswak (MIS) Zinc chloride, Salvadora persica bark extract, sodium fluoride (220 ppmF) 
Listerine Total care (EO) Eucalyptol, zinc chloride, benzoic acid, sodium benzoate, methyl salicylate, thymol, sodium fluoride (220 ppmF), menthol 

Disc Diffusion Assay

Sterile filter paper disks were impregnated with different oral rinses (25 μL) or amphotericin (250 μg/mL), dried, and then placed on SDA plates inoculated with C. albicans broth culture (1 × 106 cells/mL). After 24 h of incubation at 37°C, the zones of growth inhibition were measured.

Minimum Inhibitory Concentration and Minimum Fungicidal Concentration

The minimum inhibitory concentration (MIC) was determined with modifications of a previous study [14]. In brief, SDB (100 µL) was added to the wells of a 96-well plate, followed by an equal volume of oral rinses in the first wells. Serial dilutions were prepared, and C. albicans (100 µL) was added to each well. Negative control used SDB alone, while amphotericin (2 μg/mL) served as the positive control. After 24 h of incubation at 37°C with orbital shaking (120 rpm), p-iodonitrotetrazolium violet (Sigma; INT) dissolved in PBS (40 µL of 0.6 mg/mL) was added, followed by another 24-h incubation at 37°C. The MIC was recorded as the lowest oral rinse concentration with no visible growth or color change after 24 h. The minimum fungicidal concentration (MFC) was determined following CLSI guidelines [15].

Effect of Oral Rinses on Fungal Dimorphism

The impact of oral rinses on C. albicans morphological transition was evaluated based on a previous study [16], with minor adjustments. C. albicans (1 × 106 cells/mL) was exposed to oral rinses (1/2MIC) in 24 well plates and incubated at 37°C for 24 h. Controls consisted of C. albicans exposed to saline and amphotericin (1/2MIC). After incubation, horse serum (20%) was added and incubated for 3 h at 37oC, followed by microscopic examination for germ tube formation.

Phospholipase Assay

Phospholipase activity was determined according to a previous study with modifications [17]. Egg yolk agar plates containing oral rinse inhibitory concentrations (MIC) and control plates were prepared. C. albicans (1 × 106 cells/mL) was spot-inoculated on the agar and incubated at 37°C for 48 h. Precipitation zones around colonies indicated phospholipase activity. The phospholipase zone (Pz) index was calculated by dividing colony diameter by the total diameter plus the precipitation zone. A Pz value of 1 indicated no phospholipase activity, while values of 0.9–0.99 were considered weak, 0.89–0.70 moderate and 0.69 strong phospholipase activity [17].

Hemolysin Production

Hemolysin activity was assessed on sheep blood agar plates with oral rinse inhibitory concentrations (MIC) in a similar way as described above for the phospholipase assay. The Hz index indicated varying levels of hemolytic activity [18].

Biofilm Formation

Equal volumes (100 µL) of oral rinses or amphotericin at sub-inhibitory concentrations (1/2, 1/4, 1/8, and 1/16 times the MIC) and C. albicans (1 × 106 cells/mL) were added to tissue culture-treated flat-bottom 96-well plates and incubated for 48 h at 37°C with orbital shaking at 120 rpm to allow biofilm development. Controls included C. albicans without oral rinse or amphotericin. After biofilm formation, the medium was removed, and non-adherent cells washed away with sterile PBS. The plates were dried, stained with crystal violet (0.4%), washed and treated with 95% ethanol. The absorbance was measured at 570 nm using a microplate reader.

Mature Biofilm Disruption

For the oral rinses that did not inhibit biofilm formation at sub-inhibitory concentration, the effects on preformed mature biofilms of C. albicans were also investigated. C. albicans (1 × 106 cells/mL, 100 μL) was incubated for 48 h to allow biofilm formation. After washing, oral rinses (MIC, 2MIC, 4MIC, 8MIC, and 16MIC) were added and incubated for another 48 h. Biofilms were then quantified using crystal violet staining.

Statistical Analysis

Results were expressed as means and standard deviation values and analyzed using one- or two-way analysis of variance (ANOVA) with Minitab 21 statistical software package. Differences between control and test groups were analyzed using Tukey’s pairwise comparisons. A value of p < 0.05 was considered significant.

Table 1 shows the primary active components of the oral rinses used in the current study. The results of the disk diffusion assay indicated significant anti-Candida effects of each oral rinse, with differing sizes of growth inhibitory zones (p < 0.001, one-way ANOVA) (Fig. 1). Amphotericin exhibited the highest degree of inhibition, followed closely by the CPC-based and CHX-based rinses, which were approximately 14% less active but still significantly more effective than MIS and EO-based rinses, showing only 52% activity compared to amphotericin (Tukey’s pairwise comparison). The MIC and MFC of the oral rinses and amphotericin were also determined Table 2 and Figure 2. The most active oral rinse was based on CPC (0.2% MIC), followed by CHX (0.78% MIC), MIS, and EO (6.25% MIC for both) and a similar pattern was observed with the MFC values.

Fig. 1.

Effect of the different oral rinses on the growth of C. albicans using the disc diffusion assay. Bars represent means of the growth inhibitory zone ± SD. Means that do not share a letter are significantly different from each other (p < 0.001, one-way ANOVA; Tukey’s pairwise comparisons).

Fig. 1.

Effect of the different oral rinses on the growth of C. albicans using the disc diffusion assay. Bars represent means of the growth inhibitory zone ± SD. Means that do not share a letter are significantly different from each other (p < 0.001, one-way ANOVA; Tukey’s pairwise comparisons).

Close modal
Table 2.

Antifungal activity of oral rinses

Oral rinseMIC, %MFC, %
CHX 0.78 0.78 
CPC 0.20 0.39 
MIS 6.25 6.25 
EO 6.25 6.25 
Oral rinseMIC, %MFC, %
CHX 0.78 0.78 
CPC 0.20 0.39 
MIS 6.25 6.25 
EO 6.25 6.25 

CHX, chlorhexidine; CPC, cetylpyridinium chloride; MIS, zinc chloride and sodium fluoride-based rinses that contained Salvadora persica bark extract; EO, zinc chloride and sodium fluoride that contained the essential oils eucalyptol, thymol, and menthol (EO); MIC, minimum inhibitory concentration; MFC, minimal fungicidal concentration.

Fig. 2.

MIC of the oral rinses against C. albicans using INT as an indicator of growth. Rows A-D show the results of exposure of C. albicans to serial dilutions of the different oral rinses (50% [v/v] initial concentration). Controls include rows: E (2 μg/mL amphotericin-AMP); F (broth); G (C. albicans). Red color: viable cells; yellow color: dead cells.

Fig. 2.

MIC of the oral rinses against C. albicans using INT as an indicator of growth. Rows A-D show the results of exposure of C. albicans to serial dilutions of the different oral rinses (50% [v/v] initial concentration). Controls include rows: E (2 μg/mL amphotericin-AMP); F (broth); G (C. albicans). Red color: viable cells; yellow color: dead cells.

Close modal

Statistically significant differences were observed in germ tube production in the different treatment groups (p < 0.001, one-way ANOVA) (Fig. 3). Oral rinses containing phytochemicals (MIS and EO) exhibited the highest activity in inhibiting germ tube production (70% inhibition), with significantly more potent effects than those based on CHX (25% inhibition) and CPC (12% inhibition). Surprisingly, their impact on germ tube production was not significantly different from that of amphotericin (82% inhibition). Moreover, all oral rinses demonstrated significantly greater reductions in germ tube production when compared to the control, with one exception being the CPC-based oral rinse.

Fig. 3.

Effect of the different oral rinses on germ tube production of C. albicans exposed to the oral rinses at sub-inhibitory concentrations (1/2MIC). Controls consisted of C. albicans without the oral rinse and amphotericin (1/2MIC). The percentage of germ tube formation is shown as means ± SD. Means that do not share a letter are significantly different from each other (p < 0.0005, one-way ANOVA; Tukey’s pairwise comparisons).

Fig. 3.

Effect of the different oral rinses on germ tube production of C. albicans exposed to the oral rinses at sub-inhibitory concentrations (1/2MIC). Controls consisted of C. albicans without the oral rinse and amphotericin (1/2MIC). The percentage of germ tube formation is shown as means ± SD. Means that do not share a letter are significantly different from each other (p < 0.0005, one-way ANOVA; Tukey’s pairwise comparisons).

Close modal

We also investigated the effects of the oral rinses and amphotericin at sub-inhibitory concentrations (1/2MIC, 1/4MIC, 1/8MIC, 1/16MIC) on biofilm formation of C. albicans (Fig. 4). A two-way ANOVA was performed to analyze the effect of the concentration and type of oral rinse formulation on biofilm formation. The results showed statistically significant relations between the effects of concentration and type of oral rinse, as well as the interaction between concentration and type of oral rinse (p < 0.001 for all) on biofilm formation. Further analysis (one-way ANOVA and Tukey’s pairwise comparison) showed statistically significant inhibition of biofilm formation at all the sub-inhibitory concentration of the oral rinses tested (p < 0.001), with the greatest inhibition of biofilm formation occurring with oral rinses based on CPC, MIS, and EO, all of which caused similar levels of inhibition to amphotericin. The CHX-based oral rinse caused a small but significant inhibition at the highest concentration used, in comparison to the control. The CPC-based and phytochemical-based oral rinses showed the greatest decreases at all concentrations used and, interestingly, caused significantly greater reductions in biofilm production than the antifungal amphotericin. The CHX-based oral rinse (MIC, 2MIC, 4MIC, 8MIC, and 16MIC) had no discernible effects on the disruption of mature preformed biofilms (online suppl. Figure 1; for all online suppl. material, see https://doi.org/10.1159/000538368). This absence of impact persisted even at concentrations as high as 16 times the inhibitory concentration (online suppl. Figure 1). The impact of the oral rinses on phospholipase and hemolysin production of C. albicans is depicted in Table 3 and Figure 5. MIS and EO-based oral rinses caused total inhibition of phospholipase and hemolysin production, whereas those based on CHX and CPC exerted no effects.

Fig. 4.

Effect of the different oral rinses on biofilm formation of C. albicans exposed to the oral rinses or amphotericin at sub-inhibitory concentrations (1/2MIC). Controls consisted of C. albicans without the oral rinse. Biofilm formation was quantified using crystal violet staining and expressed as percentage of controls. Bars represent means ± SD. Means that do not share a letter are significantly different from each other (p < 0.001, one-way ANOVA; Tukey’s pairwise comparisons).

Fig. 4.

Effect of the different oral rinses on biofilm formation of C. albicans exposed to the oral rinses or amphotericin at sub-inhibitory concentrations (1/2MIC). Controls consisted of C. albicans without the oral rinse. Biofilm formation was quantified using crystal violet staining and expressed as percentage of controls. Bars represent means ± SD. Means that do not share a letter are significantly different from each other (p < 0.001, one-way ANOVA; Tukey’s pairwise comparisons).

Close modal
Table 3.

Effects of oral rinses on phospholipase and hemolysin production of C. albicans

TreatmentPzPhospholipase production*HzHemolysin production
Control 0.404±0.011 ++++ 0.402±0.012 ++++ 
Gar Garol 0.399±0.016 ++++ 0.408±0.012 ++++ 
Colgate Plax 0.408±0.015 ++++ 0.415±0.014 ++++ 
Listerine Miswak 1.000±0.000 − 1.000±0.000 − 
Listerine Total care 1.000±0.000 − 1.000±0.000 − 
TreatmentPzPhospholipase production*HzHemolysin production
Control 0.404±0.011 ++++ 0.402±0.012 ++++ 
Gar Garol 0.399±0.016 ++++ 0.408±0.012 ++++ 
Colgate Plax 0.408±0.015 ++++ 0.415±0.014 ++++ 
Listerine Miswak 1.000±0.000 − 1.000±0.000 − 
Listerine Total care 1.000±0.000 − 1.000±0.000 − 

Pz, phospholipase; Hz, hemolysin zone index.

*Degree of production: Pz or Hz 1 = negative; <0.70 = ++++.

Fig. 5.

Phospholipase activity (a) and hemolysin activity (b) of C. albicans was determined using egg yolk agar plates or blood agar plates, respectively, containing inhibitory concentrations of the oral rinses (MIC) or without oral rinses for the controls. Note the complete inhibition of phospholipase activity (absence of precipitation zones) and hemolysis with MIS and EO while CHX and CPC demonstrated no impact.

Fig. 5.

Phospholipase activity (a) and hemolysin activity (b) of C. albicans was determined using egg yolk agar plates or blood agar plates, respectively, containing inhibitory concentrations of the oral rinses (MIC) or without oral rinses for the controls. Note the complete inhibition of phospholipase activity (absence of precipitation zones) and hemolysis with MIS and EO while CHX and CPC demonstrated no impact.

Close modal

Oral rinses offer broad-spectrum antimicrobial effects throughout the oral cavity, reaching inaccessible areas and eliminating potential microbial reservoirs that persist despite mechanical cleaning methods [19]. With so many different oral rinses that are based on a variety of natural and/or synthetic components available, oral healthcare professionals must provide evidence-based recommendations to patients for controlling microbial proliferation and inhibiting biofilm formation, thereby reducing the risk of related oral diseases.

The antifungal activity of oral rinses has been reported [13, 20], but we investigated further their effects on the key virulence factors of C. albicans, which are important for colonization, establishment, and invasion under suitable predisposing conditions. By targeting virulence factor production, oral rinses could debilitate this opportunistic pathogen from causing infection. We examined the effects on germ tube production allowing invasion of epithelial cells [6] and undermining phagosome microbicidal action [21], adhesion and biofilm formation allowing cell attachment and persistence of infection with increasing resistance to antimicrobials and host immune responses [10], and production of phospholipase and hemolysin that promote the continuous presence of C. albicans in the host [7‒9].

All tested oral rinses inhibited the growth of C. albicans, but the fungicidal effects varied according to the composition of the active ingredients. Rinses containing CPC (0.075%) and CHX (0.075%) showed the greatest fungicidal activity with the lowest MFCs (0.38% and 0.78%, respectively). Surprisingly, CPC-based rinses had greater activity than CHX, typically considered the gold standard, and these findings align with other studies [13, 22]. Conversely, zinc chloride and sodium fluoride-based rinses, including Salvadora persica bark extract (MIS) or essential oils (EO), showed similar antifungal activity (MFCs of 6.25%) but were 8–16 times less effective than CPC and CHX-based rinses. This difference may stem from the potent antimicrobial nature of CPC and CHX, effectively disrupting microbial cell membranes, compared to phytochemical-based rinses.

In immunosuppressed patients, the typically commensal C. albicans can transition to a pathogenic state, with potential life-threatening invasive infections despite chemotherapy [3, 4]. Infections develop after induction of different factors of virulence in C. albicans as it perceives changes in its microenvironment and adjusts its gene transcription profile, notably transitioning to the hyphal morphology, crucial for adhesion, invasion, tissue damage, and biofilm formation [6, 21, 23, 24]. EO-based and MIS-based oral rinses had the most substantial inhibitory effects on germ tube production, reducing it by 86–89%, surpassing CHX (42% reduction) and CPC (29% reduction). This aligns with previous findings indicating that EO components like eucalyptol completely inhibit the yeast-to-hyphal transition [25]. This observed suppression of germ tube production may be ascribed to the capacity of phytochemical-based oral rinses to disrupt pivotal signal transduction pathways or essential enzymes crucial for this process. Moreover, recent research underscores how specific constituents within essential oils can induce destabilization of cellular microtubules, which play a crucial role in the morphogenesis of fungal hyphae [26]. Similarly, CHX, though antimicrobial, showed lesser germ tube suppression [27]. Thus, while CPC and CHX oral rinses possess stronger antimicrobial properties, they exhibit inferior efficacy in inhibiting germ tube production compared to phytochemical-based rinses.

Biofilm formation not only enhances the resistance of Candida to antifungal agents but also enables the evasion of host immune defenses, increasing the risk of disseminated systemic infection [10]. The tested oral rinses significantly inhibited biofilm formation at all sub-inhibitory concentrations. While CHX displayed only minor but significant inhibition at the highest concentration tested (1/2MIC), consistent with previous findings [28], CPC and phytochemical-based rinses showed substantial reductions in biofilm formation at all concentrations, even surpassing amphotericin [29]. Phytochemical-based rinses demonstrated superior anti-biofilm activity, potentially due to their potent inhibition of germ tube formation, a pivotal step in biofilm development [28]. Conversely, the mechanism of action for CPC may involve various factors such as cell membrane disruption, adhesion prevention, inhibition of extracellular matrix production, and potential interference with quorum sensing mechanisms crucial for biofilm formation [28]. This study underscores the diverse mechanisms by which oral rinses target Candida biofilms, with CPC-based and phytochemical-based rinses exhibiting remarkable efficacy in biofilm inhibition compared to CHX-based ones [28].

We also observed significant variations in the ability of oral rinses to inhibit the release of phospholipase and hemolysin in C. albicans with no previous studies exploring this effect. Notably, only the phytochemical-based oral rinses demonstrated the ability to inhibit the release of these extracellular enzymes, suggesting that phytochemical-based oral rinses may interfere with critical mechanisms that enable C. albicans to establish and maintain infections. This underscores the therapeutic potential of phytochemical-based formulations in modulating C. albicans virulence factors, offering promising avenues for managing oral fungal infections. Furthermore, the lack of effects of CPC-based and CHX-based oral rinses on the release of enzyme highlights unique properties of phytochemical formulations in targeting these virulence factors specifically.

Our study reveals that oral rinses based on zinc chloride and sodium fluoride that contained either S. persica bark extract (MIS) or the EOs eucalyptol, thymol, and menthol had lower fungicidal activity than the CPC-based and CHX-based oral rinses but greater anti-virulence effects. For overall bacterial load reduction and promotion of oral hygiene, CHX and CPC rinses, targeting a broad spectrum of microorganisms, are beneficial. However, for addressing specific oral infections or combating the pathogenic properties of microbes like C. albicans, targeting virulence factors may be more appropriate. By inhibiting these factors crucial for colonization and invasion, the oral rinses exert a crippling effect on C. albicans, weakening this opportunistic pathogen and hindering its potential to cause infection.

We must bear in mind that the lack of inclusion of clinical isolates or non-albicans Candida species limits the wider relevance of our results. In order to circumvent this limitation, future studies should incorporate oral isolates of C. albicans and non-albicans species, together with strains that are resistant to conventional antimycotic agents. The results of such studies would enhance our knowledge of the clinical implications associated with different oral rinse formulations.

Oral rinses significantly affect C. albicans, influencing germ tube production, biofilm formation, and release of the extracellular enzyme phospholipase and hemolysin. Zinc chloride and sodium fluoride-based rinses containing phytochemicals have less fungicidal activity than CPC-based and CHX-based rinses but show stronger anti-virulence effects. However, further studies are warranted to validate these findings with other C. albicans and non-albicans Candida species and explore their broader implications for oral health.

Ethics approval is not required as this study did not include human participants or animals.

The authors have no conflicts of interest to declare.

This study was supported by the Deanship of Research at Jordan University of Science and Technology (Grant Number No. 20220521).

Homa Darmani and Dua’a Al-Saleh designed the experiments. Homa Darmani analyzed the data and together with Dua’a Al-Saleh prepared the manuscript.

Data are available on request from the corresponding author.

1.
Millsop
JW
,
Fazel
N
.
Oral candidiasis
.
Clin Dermatol
.
2016
;
34
(
4
):
487
94
.
2.
Lopes
JP
,
Lionakis
MS
.
Pathogenesis and virulence of Candida albicans
.
Virulence
.
2022
;
13
(
1
):
89
121
.
3.
Pfaller
MA
,
Diekema
DJ
.
Epidemiology of invasive mycoses in North America
.
Crit Rev Microbiol
.
2010
;
36
:
1
53
.
4.
Pappas
PG
,
Lionakis
MS
,
Arendrup
MC
,
Ostrosky-Zeichner
L
,
Kullberg
BJ
.
Invasive candidiasis
.
Nat Rev Dis Primers
.
2018
;
4
:
18026
.
5.
Lewis
MAO
,
Williams
DW
.
Diagnosis and management of oral candidosis
.
Br Dent J
.
2017
;
223
(
9
):
675
81
.
6.
Gow
NA
.
Germ tube growth of Candida albicans
.
Curr Top Med Mycol
.
1997
;
8
(
1–2
):
43
55
.
7.
Watanabe
T
,
Takano
M
,
Murakami
M
,
Tanaka
H
,
Matsuhisa
A
,
Nakao
N
, et al
.
Characterization of a haemolytic factor from Candida albicans
.
Microbiology
.
1999
;
145 (Pt 3)
:
689
94
.
8.
Ghannoum
MA
.
Potential role of phospholipases in virulence and fungal pathogenesis
.
Clin Micro Rev
.
2000
;
13
(
1
):
122
43
.
9.
Mogavero
S
,
Höfs
S
,
Lauer
AN
,
Müller
R
,
Brunke
S
,
Allert
S
, et al
.
Candidalysin is the hemolytic factor of Candida albicans
.
Toxins
.
2022
;
14
(
12
):
874
.
10.
Nobile
CJ
,
Johnson
AD
.
Candida albicans biofilms and human disease
.
Annu Rev Microbiol
.
2015
;
69
:
71
92
.
11.
Haffajee
AD
,
Yaskell
T
,
Socransky
SS
.
Antimicrobial effectiveness of an herbal mouth rinse compared with an essential oil and a chlorhexidine mouth rinse
.
J Am Dent Assoc
.
2008
;
139
(
5
):
606
11
.
12.
de Oliveira
JR
,
Belato
KK
,
de Oliveira
FE
,
Jorge
AOC
,
Camargo
SEA
,
de Oliveira
LD
.
Mouthwashes: an in vitro study of their action on microbial biofilms and cytotoxicity to gingival fibroblasts
.
Gen Dent
.
2018
;
66
(
2
):
28
34
.
13.
Nordin
R
,
Roslan
MA
,
Fathilah
AR
,
Ngui
R
,
Musa
S
.
Evaluation of in vitro antifungal effects of synthetic and herbal mouth rinses on oral Candida albicans and Candida glabrata
.
Trop Biomed
.
2022
;
39
(
3
):
302
14
.
14.
Weseler
A
,
Geiss
HK
,
Saller
R
,
Reichling
J
.
A novel colorimetric broth microdilution method to determine the minimum inhibitory concentration (MIC) of antibiotics and essential oils against Helicobacter pylori
.
Pharmazie
.
2005
;
60
(
7
):
498
502
.
15.
CLS
. In:
Wayne
P
, editor.
Reference method for broth dilution antifungal susceptibility testing of yeasts
. 4th ed.
Clinical and Laboratory Standards Institute
;
2017
.
16.
Ellepola
ANB
,
Samaranayake
LP
.
The effect of limited exposure to antimycotics on the relative cell-surface hydrophobicity and the adhesion of oral Candida albicans to buccal epithelial cells
.
Arch Oral Biol
.
1998
;
43
(
11
):
879
87
.
17.
Kantarcioglu
AS
,
Yucel
A
.
Phospholipase and protease activities in clinical Candida isolates with reference to the sources of strains
.
Mycoses
.
2002
;
45
(
5–6
):
160
5
.
18.
Budzyńska
A
,
Sadowska
B
,
Więckowska-Szakiel
M
,
Różalska
B
.
Enzymatic profile, adhesive and invasive properties of Candida albicans under the influence of selected plant essential oils
.
Acta Biochim Pol
.
2014
;
61
(
1
):
115
21
.
19.
Collins
LMC
,
Dawes
C
.
The surface area of the adult human mouth and thickness of the salivary film covering the teeth and oral mucosa
.
J Dent Res
.
1987
;
66
(
8
):
1300
2
.
20.
Ardizzoni
A
,
Pericolini
E
,
Paulone
S
,
Orsi
CF
,
Castagnoli
A
,
Oliva
I
, et al
.
In vitro effects of commercial mouthwashes on several virulence traits of Candida albicans, viridans streptococci and Enterococcus faecalis colonizing the oral cavity
.
PLoS One
.
2018
;
13
:
e0207262
.
21.
Westman
J
,
Moran
G
,
Mogavero
S
,
Hube
B
,
Grinstein
S
.
Candida albicans hyphal expansion causes phagosomal membrane damage and luminal alkalinization
.
mBio
.
2018
;
9
(
5
):
e01226
18
.
22.
Fathilah
AR
,
Himratul-Aznita
WH
,
Fatheen
ARN
,
Suriani
KR
.
The antifungal properties of chlorhexidine digluconate and cetylpyrinidinium chloride on oral Candida
.
J Dent
.
2012
;
40
(
7
):
609
15
.
23.
Hube
B
.
From commensal to pathogen: stage- and tissue-specific gene expression of Candida albicans
.
Curr Opin Microbiol
.
2004
;
7
(
4
):
336
41
.
24.
Jacobsen
ID
,
Wilson
D
,
Wächtler
B
,
Brunke
S
,
Naglik
JR
,
Hube
B
.
Candida albicans dimorphism as a therapeutic target
.
Expert Rev Anti Infec Ther
.
2012
;
10
(
1
):
85
93
.
25.
Gupta
P
,
Pruthi
V
,
Poluri
KM
.
Mechanistic insights into Candida biofilm eradication potential of eucalyptol
.
J Appl Microbiol
.
2021
;
131
(
1
):
105
23
.
26.
Shahina
Z
,
Yennamalli
RM
,
Dahms
TES
.
Key essential oil components delocalize Candida albicans Kar3p and impact microtubule structure
.
Microbiol Res
.
2023
;
272
:
127373
.
27.
Ellepola
ANB
,
Joseph
BJ
,
Khan
ZU
.
Effects of subtherapeutic concentrations of chlorhexidine gluconate on germ tube formation of oral Candida
.
Med Prin Pract
.
2012
;
21
(
2
):
120
4
.
28.
Dudek-Wicher
R
,
Junka
AF
,
Migdał
P
,
Korzeniowska-Kowal
A
,
Wzorek
A
,
Bartoszewicz
M
.
The antibiofilm activity of selected substances used in oral health prophylaxis
.
BMC Oral Health
.
2022
;
22
(
1
):
509
.
29.
Paulone
S
,
Malavasi
G
,
Ardizzoni
A
,
Orsi
CF
,
Peppoloni
S
,
Neglia
RG
, et al
.
Candida albicans survival, growth and biofilm formation are differently affected by mouthwashes: an in vitro study
.
New Microbiol
.
2017
;
40
(
1
):
45
52
.