Basic Research—Biology
Biofilm Formation Capability of Enterococcus faecalis
Cells in Starvation Phase and Its Susceptibility to Sodium
Hypochlorite
Hongyan Liu, MS, Xi Wei, MDS, PhD, Junqi Ling, PhD, Weilu Wang, MS, and Xiangya Huang, MS
Abstract
Introduction: Enterococcus faecalis is commonly
associated with persistent periapical infections. The
physiologic state of the cells in the canal is probably
closest to the starvation state. However, the biofilm
formation capability of starved E. faecalis cells on
human dentin and the susceptibility of the biofilm to
5.25% sodium hypochlorite remain poorly understood.
Methods: E. faecalis American Type Culture Collec-
tion (ATCC) 29212 in different growth phases were
incubated on human dentin and polystyrene blocks.
Scanning electron microscopy and biofilm assay were
used to investigate the biofilm formation capability of
these cells. The susceptibility of the biofilm to 5.25%
NaClO was also determined by the plate count method.
Results: Scanning electron microscopy and biofilm
assay showed that starved cells were able to form bio-
film on dentin with reduced efficiency as compared
with the cells in the exponential phase and stationary
phase (p < 0.05). Biofilm grown on dentin harbored
more cells than polystyrene (p < 0.05). Biofilms of
starved cells were more resistant to 5.25% NaClO
than those of stationary cells (p < 0.05), and the impact
of 5.25%NaClO on them decreased as the biofilm
matured. Conclusion: E. faecalis cells in the starvation
phase could develop biofilm on human dentin, which is
responsive to 5.25% NaClO. It may contribute to the
predominant role of E. faecalis involved in persistent
periapical infections. (J Endod 2010;36:630–635)
Key Words
Biofilm, dentin, Enterococcus faecalis, sodium
hypochlorite
From the Department of Operative Dentistry and Endodon-
tics, Guanghua School of Stomatology, Sun Yat-sen University,
Guangzhou, Guangdong, China.
*Hongyan Liu and Xi Wei contributed equally to this study.
Supported by Key Clinical Program of the Ministry of
Health, China (no. [2007] 353).
Address requests for reprints to Dr Junqi Ling, Department
of Operative Dentistry and Endodontics, Guanghua School of
Stomatology, Sun Yat-sen University, Guangzhou 510055, PR
China. E-mail address: lingjq@mail.sysu.edu.cn.
0099-2399/$0 - see front matter
Copyright ª 2010 American Association of Endodontists.
doi:10.1016/j.joen.2009.11.016
630 Liu et al.
E nterococcus faecalis is a predominant organism that plays a major role in the
etiology of persistent periradicular lesions after root canal treatment (1, 2).
Studies investigating its occurrence in root-filled teeth with periradicular lesions
have shown a prevalence ranging from 24% to 77% (3). The inherent antimicrobial
resistance and the ability to adapt to changing environment help E. faecalis to persist
in root canal.
E. faecalis can adhere to the root canal walls, accumulate, and form communities
organized in biofilm, which helps it resist destruction by enabling the bacteria to
become 1,000 times more resistant to phagocytosis, antibodies, and antimicrobials
than non–biofilm-producing organisms (3). In vitro studies have shown a similar bio-
film formation capability of E. faecalis strains isolated from oral cavity and root canals
(4) and the ability of E. faecalis biofilm to calcify as it undergoes ‘‘maturation’’ within
the root canal (5).
Most of the studies on E. faecalis biofilm formation in root canal have been con-
ducted in environmental conditions allowing cells to grow and divide normally (6, 7).
However, it has been shown that a major change in the environment induced by root
canal treatment is related to a dramatic reduction in nutrient availability and oxygen
(8). Bacterial cells in the root canal encounter a harsh ecologic milieu. It is probable
that the physiologic state of the cells in the canal, particularly in retreatment cases, is
closest to the starvation phase (9).
There have been few studies on the biofilm formed by starved E. faecalis cells on
dentin. George et al (10) showed distinct ultrastructure of the E. faecalis biofilms
formed under nutrient-rich, nutrient-deprived, aerobic, and anaerobic conditions.
They also reported a significant increase in the level of calcium in the biofilm structures
formed under an anaerobic nutrient-deprived condition and a significantly greater
depth of bacterial penetration in nutrient-rich condition. Lleo et al (11) reported
that starved and stationary enterococcal cells were able to form biofilms on plastic mate-
rial albeit with reduced efficiency as compared with cells in the exponential phase.
However, particular environmental stimuli such as the substratum could trigger
different developmental pathways and influence the quantities although not the struc-
ture of the associated biofilms (12). By far, the topographic changes in E. faecalis bio-
film developed by starved cells on human dentin remain poorly understood.
Furthermore, the comparative study of the biofilm formation ability of starved cells
on dentin with the cells in exponential phase and stationary phase has not been re-
ported.
The physiologic state of the bacteria will also have an effect on the outcome of the
antimicrobial treatment. For planktonic cultures, starvation has been shown to increase
resistance in E. faecalis cells to chemical, osmotic, and oxidative stress (13). Portenier
et al (9) reported that starved cells survive in numbers 1,000 to 10,000 times higher
after being challenged with sodium hypochlorite, one of the most commonly used
endodontic irrigants, as compared with cells in the exponential phase or stationary
phase. However, the impact of NaClO on biofilms formed by starved cells has not yet
been investigated.
In this study, the ability to form biofilms of starved E. faecalis cells was compared
with those in exponential and stationary phases. The susceptibility of the so-formed bio-
film to 5.25% NaClO was also investigated.
JOE — Volume 36, Number 4, April 2010

Material and Methods
Bacterial Strain, Culture Conditions, and Starvation
Induction
E. faecalis (ATCC 29212; Guangdong Provincial Key Laboratory of
Microbiol Culture Collection and Application, Guangdong Institute
of Microbiology, Guangzhou, China) was used in this study; 500 mL
of bacterium (1.5  107 CFU/mL) were added to 50 mL brain heart
infusion (BHI) medium, which was further incubated without shaking
at 37 C under anaerobic condition. Cell growth were monitored by the
colony-forming unit (CFU) counting of the culturable bacteria on BHI
agar plate as described by Portenier et al (9). In detail, after the desig-
nated incubation time, 1 mL of bacterial suspension was serially diluted
and plated on BHI agar in triplicate. After 48 hours of incubation in an
anaerobic atmosphere at 37 C, the number of colonies was counted,
yielding a value in CFU/mL.
For the induction of starvation state, cells in the exponential phase
(1.5  109 CFU/mL, after incubation for 4 hours) were harvested,
washed twice with 10 mmol/L phosphate-buffered saline (PBS, pH =
7), resuspended in PBS at a final density of 109 CFU/mL, and maintained
at 37 C for 48 hours without direct illumination (14). Colony-forming
ability was determined in duplicate on samples plated on BHI agar plate.
Specimen Preparation
Healthy human third molars were extracted from adults (19-25
years old) in the Department of Oral and Maxillofacial Surgery, Sun
Yat-sen University Dental Hospital, Guangzhou, China, after obtaining
informed consent from each subject, and the protocols were approved
by the University Ethic Committee. The teeth were split longitudinally,
and the coronal pulp chamber surfaces were prepared into blocks
(4.5 mm  4.5 mm  1 mm) without scraping. The same size of poly-
styrene (PLS) blocks were made. All blocks were washed with distilled
water and ultrasonically cleaned. Dentin blocks were autoclaved. PLS
blocks were sterilized by ultraviolet irradiation. All sterile blocks
were stored in 10 mmol/L PBS at 4 C and used within 1 week.
Biofilm Assay
Bacterial cells in the exponential, stationary (after incubation for
12 hours), and starvation phase were harvested and prepared at the
same concentration of 1  108 CFU/mL. BHI medium, the nutrient-
depleted BHI medium, and 10 mmol/L PBS were prepared as culture
medium for biofilm formation of cells in the exponential, stationary,
and starvation phase, respectively. Nutrient-depleted BHI medium
was prepared as described before (15) with some modifications. In
detail, the medium was prepared by adding E. faecalis to BHI medium
and incubating at 37 C under anaerobic condition for 10 hours. After-
ward, the cultures were centrifuged (5,000 rpm, 10 minutes) at 4 C to
acquire the supernatant. The pH of the supernatant was adjusted to pH
7.0, and then the medium was passed through a 0.22-mm-pore-size
filter. Blocks were placed in 24-well polystyrene cell culture plates
with pulpal surfaces upwards for dentin blocks. Each well were inocu-
lated with a suspension of E. faecalis cells (1 mL of 108CFU/mL) and
plates were left undisturbed in anaerobic incubator at 37 C for 6 hours,
24 hours, 48 hours, 72 hours, 96 hours, or 168 hours. The bacterial
suspension was replaced every day. In detail, the cultures containing
nonadherent cells in the wells were aspired. Fresh BHI medium,
nutrient-depleted BHI medium, or PBS was added to the 24-well culture
plate with dentin or PLS blocks. All biofilm assays were performed in
triplicate in at least three independent experiments on different days.
At the end of incubation time, each block was aseptically removed
and washed twice by being kept in PBS and shaking on a shaker (Uni-
twist RT, Uniequip Company, Laborgerätebau & vertriebs GmbH, Mar-
JOE — Volume 36, Number 4, April 2010
Basic Research—Biology
tinsried, Germany) for 30 seconds to remove loosely adherent cells. The
blocks were subsequently kept in 1 mL of cysteine peptone water (16)
for 1 minute. Biofilm samples were harvested by scraping and vigorous
pipetting. The bacteria were serially diluted. Each dilution was plated
onto BHI plates. The plates were then incubated for 48 hours in an
anaerobic atmosphere at 37 C, and CFU per block were calculated.
Sodium Hypochlorite Treatment
A solution of 5.25% (m/v) sodium hypochlorite (NaClO) was
freshly prepared. Forty-eight-hour-old E. faecalis biofilm formed by
bacterial cells in different physiologic states on dentin blocks were
removed aseptically from the wells; washed twice with 10 mmol/L
PBS; transferred into 1 mL of 5.25% NaClO; and incubated for 30
seconds, 10 minutes, and 30 minutes, respectively. After the designated
contact time, the blocks were carefully transferred to 1 mL of 0.6%
sodium thiosulfate to terminate the antimicrobial action of NaClO
(17). Then, the blocks were transferred into 1 mL of cysteine peptone
water and the suspensions were harvested, serially diluted, and quanti-
fied as described earlier. Controls were exposed to 10 mmol/L PBS
instead of 5.25% NaClO. All NaClO treatments were performed in trip-
licate in three independent experiments.
In order to investigate the impact of 5.25% NaClO on biofilms at
different stages of maturation, 6-hour-old, 24-hour-old, and
48-hour-old biofilms developed by E. faecalis cells in the starvation
phase were exposed to 5.25% NaClO for 30 seconds, 10 minutes,
and 30 minutes, respectively. Bacterial viability was determined as
described previously.
Scanning Electron Microscopy
After the designed incubation time, dentin and PLS blocks with bi-
ofilm were washed twice with 10 mmol/L PBS buffer and fixed with 2 mL
of 2.5% glutaraldehyde for 2 hours at 4 C. Further preparations
included dehydration through a series of ethanol rinses (30%, 50%,
70%, 95%, and 100%) and critical point drying with liquid CO2 (for
PLS blocks, air dried) before coating with gold (Hitachi E-1010, Ibrar-
aki, Japan) with the thickness of approximately 20 nm. Scanning elec-
tron micrographs of at least three randomly selected areas from each of
the three samples were taken with a scanning electron microscope
operating at 20 kV (Hitachi S-3000 N). Images presented in the
following results were representative of three samples.
Statistical Analysis
Statistical analysis was performed using SPSS software (SPSS for
windows; SPSS Inc, Chicago, IL). For the effect of substratum and phys-
iologic phase on the biofilm formation capability of E. faecalis during
different phases of maturation of biofilm, the data of CFU counts were
analyzed with analysis of variance for a 3  2  6 factorial design.
For the comparative study of susceptibility of biofilm developed by
E. faecalis cells in different physiologic states to 5.25% NaClO and
the impact of biofilm aging on the susceptibility of biofilm formed by
starved E. faecalis cells, the reduction of viable cells per block were
analyzed by analysis of variance for a 3  3 factorial design followed
by the Bonferroni test; p < 0.05 was regarded as the difference of statis-
tical significance. Data that violated the prerequisites of normal distri-
butions or equality of variances were pretreated with logarithmic
transformation.
Results
Growth of E. faecalis in BHI Medium
The amount of living cells increased exponentially during the first
6 hours of incubation (the exponential phase). Between 6 hours and 24
Biofilm Formation of E. faecalis Cells in Starvation Phase 631
Basic Research—Biology
hours, the amount of living cells remained stable (the stationary phase);
it decreased between 24 hours and 48 hours and remained stable there-
after (Fig. 1).
Ultrastructure of Biofilm Developed by Starved
E. faecalis Cells on Dentin
We used scanning electron microscopy to monitor the ultrastruc-
ture of biofilm developed by E. faecalis cells in different physiologic
phases on dentin or PLS blocks (Fig. 2). The representative micropho-
tographs showed that biofilm formation by starved cells on dentin pro-
gressed through each of the following typical biofilm stages: initial
attachment of individual cells (2 hours, Fig. 2A), microcolony forma-
tion (6 hours, 16 hours, Fig. 2B and C), and development into a mature
biofilm with complex architecture (24 hours, Fig. 2D). Thus, E. faecalis
cells in the starvation phase could form biofilms that exhibited typical
biofilm architecture on dentin. Meanwhile, E. faecalis cells in the expo-
nential (Fig. 2E) and stationary (Fig. 2F) phases developed matured bi-
ofilm on dentin after incubation for 24 hours. On PLS blocks, the
starved E. faecalis cells could also form matured biofilm at this time
point (Fig. 2G).
Biofilm Development Capability of E. faecalis in Different
Physiologic States on Human Dentin or PLS Blocks
Statistical analysis showed that there were significant interactive
effects among the substratum, physiologic phase, and incubation time
in terms of biofilm formation capacity. Both the physiologic state of cells
and substratum has significant effects on the biofilm formation capa-
bility. The CFU counts varied with the physiologic state of cells
(p < 0.001, n = 9) and the substratum (p < 0.001, n = 9) after
a different incubation time (p < 0.001, n = 9). First, the average levels
of the physiologic state of cells were significantly different, xgrowing >
xstationary > xstarvation (p < 0.05, n = 9), with the other two factors
controlled. Second, in terms of the substratum, xdentin was greater
thanxPLS (p < 0.05, n = 9) with the other two factors controlled. Third,
as for the incubation time, with the other two factors controlled, the
difference of the CFU counts of viable cells between 48 hours and 96
hours and also between 168 hours and all the other groups were statis-
tically significant (p < 0.05, n = 9). There was no statistically signifi-
cance between other groups (Fig. 3). It was shown that starved cells
were able to form biofilms with reduced efficiency as compared with
cells in exponential phase and stationary cells.
The results were further analyzed with file split by substratum. On
human dentin blocks, there was no interactive effect between the phys-
iologic state and incubation time. Over the period of incubation, the
difference in cell density of biofilm formed by E. faecalis cells in three
physiologic phases on dentin were statistically significant (p < 0.05,
n = 9) (Fig. 3B).
There was an interactive effect between the physiologic state and
incubation time on PLS blocks (p < 0.05, n = 9). Data were further
analyzed by a splitting file with incubation time. The results showed
that at different stages of biofilm maturation, the patterns of comparative
levels of the biofilm formation capability of E. faecalis cells in the expo-
nential phase, the stationary phase, and the starvation phase varied with
incubation time (Fig. 3C).
Susceptibility of Biofilm Formed by E. faecalis Cells in
Different Physiologic States to 5.25% NaClO on Human
Dentin
Forty-eight-hour-old biofilm formed by E. faecalis cells in expo-
nential, stationary, and starved phases on human dentin blocks were
exposed to 5.25% NaClO for 30 seconds, 10 minutes, and 30 minutes.
632 Liu et al.
Figure 1. Growth of E. faecalis in BHI medium; 500 mL of bacterium (1.5 
107 CFU/mL) added to 50 mL BHI medium, which was further incubated
without shaking at 37 C under anaerobic condition. Bacterial viability was
determined by plate count method. During the first 6 hours of incubation,
the cells were in exponential phase. Between 6 hours and 24 hours, the cells
were in the stationary phase.
Statistical analysis showed that there was no interactive effect between
the physiologic state and contact time. Over the period of 30 minutes,
the reduction of viable cells of biofilms developed by stationary cells
was significantly higher than those of biofilms formed by cells in the
other two phases (p < 0.05, n = 9). No significant difference was de-
tected between biofilms formed by cells in the exponential and starva-
tion phases (p > 0.05, n = 9, Fig. 4).
We investigated the effect of biofilm aging on the susceptibility
of biofilms developed by starved E. faecalis cells to 5.25% NaClO.
Statistical analysis showed that there was no interactive effect between
incubation time and contact time. Over the period of contact time,
48-hour-old biofilms showed a significant lower reduction of viable
cells compared with those of 6-hour-old and 24-hour-old biofilms
(p < 0.05, n = 9, Fig. 5).
Discussion
The results presented here showed that starved E. faecalis cells
were fully capable of biofilm formation on human dentin. The formation
of biofilm by starved E. faecalis cells on human dentin appeared to
progress through the following typical stages that had been previously
observed during biofilm development of esp-negative E. faecalis strain
in a chemostat-based biofilm fermenter (18): initial bacterial adher-
ence to the substratum, microcolony formation, and maturation into
complex three-dimensional structures.
Our results indicated that the nature of substratum may influence
the biofilm formation (12). With the other two factors (physiologic state
and incubation time) controlled, biofilm developed by E. faecalis cells
displayed higher growth on human dentin than on PLS blocks. The bio-
film formation capability of E. faecalis cells in different physiologic
states varied with the incubation time on PLS; it remained stable on
dentin. The specific binding of E. faecalis cells to dentin may account
for the consistent adherence and biofilm formation on dentin. First,
E. faecalis could invade dentinal tubules and adhere to pulpal walls
mediated by bacterial specific cell-surface components such as aggre-
gation substance (3) and Ace (collagen-binding protein of enterococci)
(19). Second, some components of dentin, such as collagen type I, may
act as selective factors for bacterial adherence and invasion (20, 21).
However, the exact interactive mechanism of E. faecalis with dentin
needs further investigation.
JOE — Volume 36, Number 4, April 2010
 Basic Research—Biology

Figure 2. SEM analysis of biofilm development by E. faecalis cells in different physiologic phases on dentin and PLS blocks (original magnification 2,000).
Aliquots of 1 mL of cell suspension of starved E. faecalis cells were grown on human dentin and incubated anerobically undisturbed at 37 C. (A) Tight
cell-substratum attachment was observed after incubation for 2 hours. After incubation for 6 hours (B) and 16 hours (C), cellular aggregation and microcolony
formation were observed. (D) Biofilm matured into complex three-dimensional structure after incubation for 24 hours. Biofilm developed by E. faecalis cells in
exponential (E) and stationary (F) phases matured after incubation for 24 hours. (G) Twenty-four-old biofilm was formed on PLS blocks by starved E. faecalis cells.

Our data showed that with factors of substratum and incubation
time controlled, the E. faecalis cells in the exponential phase formed
biofilm with higher efficiency as compared with stationary cells. Starved
cells showed the lowest biofilm formation capability. These findings
agreed with a previous study (11) in which E. faecalis cells in three
physiological states were grown on plastic material surface to form bio-
film. Interestingly, George and Kishen (22) reported that the hydropho-
bicity and adherence of E. faecalis to dentin were significantly increased
during starvation. Although adherence is considered to be the first step
for bacterial colonization of host tissue, biofilm formation is more likely

JOE — Volume 36, Number 4, April 2010 Biofilm Formation of E. faecalis Cells in Starvation Phase 633
Basic Research—Biology
Figure 3. The growth of biofilms of E. faecalis cells in different physiologic states
on various substrata over 168 hours. Bacterial cells (1 mL of 108cells/mL) in expo-
nential, stationary, and starvation phases were grown on PLS and human dentin
blocks incubated anerobically undisturbed at 37 C for 6 hours, 24 hours, 48
hours, 72 hours, 96 hours, or 168 hours. (A) Substratum has significant effect
on the biofilm formation capability at different stages of biofilm formation. The
CFU counts varied with the substratum (p < 0.001, n = 9) after different incuba-
tion time. **Significant difference between connected groups (p < 0.05). (B)
Further analysis by splitting file by substratum showed that on dentin cells in expo-
nential phase showed the highest growth followed by stationary cells. Starved cells
displayed the lowest growth. Difference among them was statistically significant
(p < 0.05, n = 9). (C) The results on PLS blocks varied with the increase of incu-
bation time, which differed from those on dentin. *Significant difference against
stationary cells (p < 0.05). #Significant difference against starved cells (p < 0.05).
634 Liu et al.
Figure 4. Susceptibility of biofilm formed by E. faecalis cells in different
physiologic states on dentin to 5.25% NaClO; 48-hour-old biofilm formed by
E. faecalis cells in exponential, stationary, and starved phases on human dentin
blocks were exposed to 5.25% NaClO for 30 seconds, 10 minutes, and 30
minutes. Statistical analysis showed that over the period of 30 minutes of
contact, biofilms of cells in exponential and starvation phases showed similar
reduction in enumeration of viable cells (p > 0.05, n = 9), which were signif-
icantly lower than that of biofilm of stationary cells (p < 0.05, n = 9).
to be dependent on cell-to-cell adhesion rather than on the amount of
cells initially attached to the surface (23). Moreover, it has been shown
that environmental signals and regulatory pathways influence biofilm
formation (24). A release of catabolite repression and the change of
fsr and its downstream proteases mediated by a glucose-dependent
transcriptional regulator (25, 26) may account for the reduced biofilm
formation capability of starved E. faecalis cells. Further study needs to
be performed to identify the signal transduction proteins involved in bi-
ofilm formation in E. faecalis.
Our study showed that E. faecalis biofilms developed by starved
cells and cells in the exponential phase were more resistant to 5.25%
NaClO than those of stationary cells. Portenier et al (9) reported that
cells harvested during the stationary phase were more resistant than
cells in the exponential phase, and starved cells were the most resistant.
We also found that the susceptibility of E. faecalis biofilm formed by
starved cells decreased with the maturation of the biofilms. Similar
observations in Actinobacillus actinomycetemcomitans biofilm and
Streptococcus mutans biofilm have also been reported by Japanese
and British researchers (27, 28).
Multiple factors may play a cumulative role in the resulting
increased biofilm resistance (29). First, microbial nutrients and waste
products modify the local environment inside a biofilm. Cells deeply
placed within the biofilm may slow their growth as the results of
some nutrient limitation. It has been suggested that the slow growth
can account for the resistance of biofilm to antimicrobial agents (29,
30). In our present studies, it is possible that E. faecalis cells in the
exponential phase in biofilm may slow their growth rate and be more
resistant to 5.25% NaClO than their ‘‘normal’’ cells. Second, biofilm/
attachment-specific phenotypes develop inside a biofilm. It has been
suggested that some genes are activated or repressed in biofilms
compared with planktonic cells (31–33). It is possible that specific
gene expression may occur during biofilm formation of E. faecalis
JOE — Volume 36, Number 4, April 2010

Figure 5. The impact of 5.25% NaClO application on biofilms at different
stages developed by starved E. faecalis cells; 6-hour-old, 24-hour-old, and
48-hour-old biofilms developed by E. faecalis cells in starvation phase were
exposed to 5.25% NaClO for 30 seconds, 10 minutes, and 30 minutes. CFU
counts of viable cells were determined by serial dilutions on BHI agar plate.
The 48-hour-old biofilms was found to be more resistant to 5.25% NaClO
than 6-hour- and 24-hour-old biofilms (p < 0.05, n = 9).
cells. Finally, the physical and/or chemical structure of exopolysacchar-
ides or other aspects of biofilm architecture confer resistance by exclu-
sion of biocides from the bacterial community (29). A study about
E. faecalis biofilm development showed that after 24 hours of growth
there was no or little exopolysaccharides, and the presence of exopoly-
saccharides uniformly distributed consolidated biofilm formation after
36 hours (34). Therefore, we speculate that the increased resistance of
48-hour-old biofilm may be an artifact partly because of the presence of
exopolysaccharides. This hypothesis needs to be verified in further
study. There are some limitations of this experiment. The sample size
is small, and there is a possibility that some of the results may not be
biologically significant; however, it is a potential suggestion that the
resistance of biofilm to 5.25% NaClO may be influenced by the physio-
logical state of cells and the aging of biofilm.
In conclusion, this study showed that E. faecalis in the starvation
phase could survive through an adverse environment and form biofilms
on human dentin with reduced efficency as compared with cells in the
exponential phase and stationary cells. The biofilms developed by
starved E. faecalis cells were more resistant to 5.25% NaClO than those
of stationary cells and similar to those of cells in the exponential phase.
The ability of starved E. faecalis to survive through a harsh environment
and form biofilms that are responsive to 5.25% NaClO may contribute to
the predominant role of E. faecalis involved in persistent periapical
infections.
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