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PLoS One. 2016 Nov 23;11(11):e0166997. doi: 10.1371/journal.pone.0166997. eCollection 2016.

Treatment of Oral Biofilms by a D-Enantiomeric Peptide.

Author information

Division of Endodontics, Department of Oral Biological and Medical Sciences, Faculty of Dentistry, University of British Columbia, Vancouver, British Columbia, Canada.
Centre for Microbial Diseases and Immunity Research, Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada.
Synthetic Biology Group, MIT Synthetic Biology Center, Research Laboratory of Electronics, Department of Biological Engineering, Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, United States of America.


Almost all dental diseases are caused by biofilms that consist of multispecies communities. DJK-5, which is a short D-enantiomeric, protease-resistant peptide with broad-spectrum anti-biofilm activity, was tested for its effect on oral multispecies biofilms. Peptide DJK-5 at 10 μg/mL effectively prevented the growth of these microbes in culture media in a time-dependent manner. In addition to the prevention of growth, peptide DJK-5 completely killed both Streptococcus mutans and Enterococcus faecalis suspended from biofilms after 30 minutes of incubation in liquid culture media. DJK-5 also led to the effective killing of microbes in plaque biofilm. The proportion of bacterial cells killed by 10 μg/mL of DJK-5 was similar after 1 and 3 days, both exceeding 85%. DJK-5 was able to significantly prevent biofilm formation over 3 days (P = 0.000). After 72 hours of exposure, DJK-5 significantly reduced and almost completely prevented plaque biofilm production by more than 90% of biovolume compared to untreated controls (P = 0.000). The proportion of dead biofilm bacteria at the 10 μg/mL DJK-5 concentration was similar for 1- and 3-day-old biofilms, whereby >86% of the bacteria were killed. DJK-5 was also able to kill >79% and >85% of bacteria, respectively, after one-time and three brief treatments of 3-day-old biofilms. The combination of DJK-5 and chlorhexidine showed the best bacterial killing among all treatments, with ~83% and >88% of bacterial cells killed after 1 and 3 minutes, respectively. No significant difference was found in the percentage of biofilm killing amongst three donor plaque samples after DJK-5 treatment. In particular, DJK-5 showed strong performance in inhibiting biofilm development and eradicating pre-formed oral biofilms compared to L-enantiomeric peptide 1018. DJK-5 was very effective against oral biofilms when used alone or combined with chlorhexidine, and may be a promising agent for use in oral anti-biofilm strategies in the future.

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Conflict of interest statement

Competing Interests: C.D.L.F.-N. and R.E.W.H. are co-inventors of a patent application on the use of cationic anti-biofilm peptides that has been licensed to ABT Innovations Inc., which is partially owned by R.E.W.H. Competing Interests Statement: R.E.W. Hancock is developing anti-biofilm peptides as new therapeutics and has a filed U.S. patent related to peptide DJK5 (Robert E. W. Hancock and César de la Fuente-Núñez, No. 61/870,655) being licensed to a virtual company ABT Innovations Inc. of which Hancock is a part owner. ABT Innovations provided no funding for this publication and had no role in study design, performance or analysis and the sequence of peptide DJK5 has been published making it freely available for research purposes. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

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