- Lactobacillus plantarum lipoteichoic acid disrupts mature Enterococcus faecalis biofilm
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A Reum Kim , Minji Kang , Yeon-Jee Yoo , Cheol-Heui Yun , Hiran Perinpanayagam , Kee-Yeon Kum , Seung Hyun Han
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J. Microbiol. 2020;58(4):314-319. Published online January 28, 2020
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DOI: https://doi.org/10.1007/s12275-020-9518-4
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Apical periodontitis is caused by biofilm-mediated root canal
infection. Early phase oral bacterial biofilms are inhibited by
Lactobacillus plantarum lipoteichoic acid (Lp.LTA). However,
mature biofilms that develop over 3 weeks are more resistant
to traditional endodontic medicaments. Therefore, this study
examined the effectiveness of Lp.LTA on disrupting mature
Enterococcus faecalis biofilms, and on enhancing the effects
of endodontic medicaments. LTA was purified from L. plantarum
through butanol extraction followed by hydrophobic
and ion-exchange chromatography. E. faecalis biofilms were
formed over 3 weeks on glass bottom dishes and in dentin
blocks obtained from human single-rooted premolars. These
mature biofilms were treated with or without Lp.LTA for 1 h,
followed by additional treatment with either chlorhexidine
digluconate (CHX), calcium hydroxide (CH), or triple antibiotics
for 24 h. Biofilms on glass were live/dead stained and
quantified by ZEN through confocal laser microscopy. Biofilms
in dentin were fixed, sputter coated and analyzed by
ImageJ with scanning electron microscopy. Preformed E. faecalis
mature biofilms on the culture dishes were dose-dependently
disrupted by Lp.LTA. Lp.LTA potentiated the effects
of CHX or CH on the disruption of mature biofilm. Interestingly,
CHX-induced disruption of preformed E. faecalis
mature biofilms was synergistically enhanced only when pretreated
with Lp.LTA. Furthermore, in the dentin block model,
Lp.LTA alone reduced E. faecalis mature biofilm and
pre-treatment with Lp.LTA promoted the anti-biofilm activity
of CHX. Lp.LTA could be an anti-biofilm or supplementary
agent that can be effective for E. faecalis-biofilminduced
diseases.
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Citations
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- Lipoteichoic acids of lactobacilli inhibit Enterococcus faecalis biofilm formation and disrupt the preformed biofilm
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Solmin Jung , Ok-Jin Park , A Reum Kim , Ki Bum Ahn , Dongwook Lee , Kee-Yeon Kum , Cheol-Heui Yun , Seung Hyun Han
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J. Microbiol. 2019;57(4):310-315. Published online January 22, 2019
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DOI: https://doi.org/10.1007/s12275-019-8538-4
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Abstract
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Enterococcus faecalis, a Gram-positive bacterium commonly
isolated in patients with refractory apical periodontitis, invades
dentin tubules easily and forms biofilms. Bacteria in biofilms,
which contribute to recurrent and/or chronic inflammatory
diseases, are more resistant to antimicrobial agents
than planktonic cells and easily avoid phagocytosis. Although
Lactobacillus plantarum lipoteichoic acid (Lp.LTA) is associated
with biofilm formation, the effect of Lp.LTA on biofilm
formation by E. faecalis is not clearly understood. In this
study, we investigated whether Lp.LTA inhibits E. faecalis
biofilm formation. The degree of biofilm formation was determined
by using crystal violet assay and LIVE/DEAD bacteria
staining. The quantification of bacterial growth was determined
by measuring the optical density at 600 nm with a
spectrophotometer. Formation of biofilms on human dentin
slices was observed under a scanning electron microscope.
E. faecalis biofilm formation was reduced by Lp.LTA treatment
in a dose-dependent manner. Lp.LTA inhibited biofilm
development of E. faecalis at the early stage without affecting
bacterial growth. LTA from other Lactobacillus species
such as Lactobacillus acidophilus, Lactobacillus casei, or
Lactobacillus rhamnosus GG also inhibited E. faecalis biofilm
formation. In particular, among LTAs from various lactobacilli,
Lp.LTA showed the highest inhibitory effect on biofilms
formed by E. faecalis. Interestingly, LTAs from lactobacilli
could remove the biofilm preformed by E. faecalis.
These inhibitory effects were also observed on the surface of human dentin slices. In conclusion, Lactobacillus species LTA
inhibits biofilm formation caused by E. faecalis and it could
be used as an anti-biofilm agent for prevention or treatment
against E. faecalis-associated diseases.
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- The synthetic human beta-defensin-3 C15 peptide exhibits antimicrobial activity against Streptococcus mutans, both alone and in combination with dental disinfectants
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Ki Bum Ahn , A Reum Kim , Kee-Yeon Kum , Cheol-Heui Yun , Seung Hyun Han
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J. Microbiol. 2017;55(10):830-836. Published online September 28, 2017
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DOI: https://doi.org/10.1007/s12275-017-7362-y
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Abstract
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Streptococcus mutans is a major etiologic agent of human
dental caries that forms biofilms on hard tissues in the human
oral cavity, such as tooth and dentinal surfaces. Human
β-defensin-3 (HBD3) is a 45-amino-acid natural antimicrobial
peptide that has broad spectrum antimicrobial activity
against bacteria and fungi. A synthetic peptide consisting of
the C-terminal 15 amino acids of HBD3 (HBD3-C15) was
recently shown to be sufficient for its antimicrobial activity.
Thus, clinical applications of this peptide have garnered
attention. In this study, we investigated whether HBD3-C15
inhibits the growth of the representative cariogenic pathogen
Streptococcus mutans and its biofilm formation. HBD3-C15
inhibited bacterial growth, exhibited bactericidal activity,
and attenuated bacterial biofilm formation in a dose-dependent
manner. HBD3-C15 potentiated the bactericidal and
anti-biofilm activity of calcium hydroxide (CH) and chlorhexidine
digluconate (CHX), which are representative disinfectants
used in dental clinics, against S. mutans. Moreover,
HBD3-C15 showed antimicrobial activity by inhibiting biofilm
formation by S. mutans and other dentinophilic bacteria
such as Enterococcus faecalis and Streptococcus gordonii,
which are associated with dental caries and endodontic
infection, on human dentin slices. These effects were observed
for HBD3-C15 alone and for HBD3-C15 in combination with
CH or CHX. Therefore, we suggest that HBD3-C15 is a potential
alternative or additive disinfectant that can be used
for the treatment of oral infectious diseases, including dental
caries and endodontic infections.
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