Anaerobe 32 (2015) 18e23

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Clinical microbiology

In-vitro activity of taurolidine on single species and a multispecies population associated with periodontitis Lilly Zollinger a, Simone Schnyder a, Sandor Nietzsche b, Anton Sculean a, Sigrun Eick a, * a b

School of Dental Medicine, Department of Periodontology, University of Bern, Switzerland Center of Electron Microscopy, University Hospital of Jena, Jena, Germany

a r t i c l e i n f o

a b s t r a c t

Article history: Received 15 September 2014 Received in revised form 14 November 2014 Accepted 16 November 2014 Available online 24 November 2014

The antimicrobial activity of taurolidine was compared with minocycline against microbial species associated with periodontitis (four single strains and a 12-species mixture). Minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs), killing as well as activities on established and forming single-species biofilms and a 12-species biofilm were determined. The MICs of taurolidine against single species were always 0.31 mg/ml, the MBCs were 0.64 mg/ml. The used mixed microbiota was less sensitive to taurolidine, MIC and the MBC was 2.5 mg/ml. The strains and the mixture were completely killed by 2.5 mg/ml taurolidine, whereas 256 mg/ml minocycline reduced the bacterial counts of the mixture by 5 log10 colony forming units (cfu). Coating the surface with 10 mg/ml taurolidine or 256 mg/ml minocycline prevented completely biofilm formation of Porphyromonas gingivalis ATCC 33277 but not of Aggregatibacter actinomycetemcomitans Y4 and the mixture. On 4.5 d old biofilms, taurolidine acted concentration dependent with a reduction by 5 log10 cfu (P. gingivalis ATCC 33277) and 7 log10 cfu (A. actinomycetemcomitans Y4) when applying 10 mg/ml. Minocycline decreased the cfu counts by 1e2 log10 cfu independent of the used concentration. The reduction of the cfu counts in the 4.5 d old multi-species biofilms was about 3 log10 cfu after application of any minocycline concentration and after using 10 mg/ml taurolidine. Taurolidine is active against species associated with periodontitis, even within biofilms. Nevertheless a complete elimination of complex biofilms by taurolidine seems to be impossible and underlines the importance of a mechanical removal of biofilms prior to application of taurolidine. © 2014 Elsevier Ltd. All rights reserved.

Keywords: Taurolidine Minocycline Periodontal pathogens Biofilm

1. Introduction Periodontitis is a bacterially induced chronic inflammatory disease, and an imbalance of innate immune-defence system markedly contributes to the destruction of the periodontium [1]. Microorganisms are organized in biofilms, the subgingival biofilms consist of hundreds of species. Bacteria more present in periodontitis than in periodontal health are Treponema denticola, Porphyromonas gingivalis, Tannerella forsythia, Aggregatibacter actinomycetemcomitans and several others [2]. P. gingivalis a gramnegative anaerobe bacterium is postulated to be a keystone pathogen in developing periodontal disease [3]. The most important * Corresponding author. University of Bern, School of Dental Medicine, Department of Periodontology, Laboratory of Oral Microbiology, Freiburgstrasse 7, CH3010 Bern, Switzerland. Tel.: þ41 31 632 25 42; fax: þ41 31 632 8608. E-mail address: [email protected] (S. Eick). http://dx.doi.org/10.1016/j.anaerobe.2014.11.008 1075-9964/© 2014 Elsevier Ltd. All rights reserved.

virulence factor of A. actinomycetemcomitans is being a leukotoxin able to cause imbalance in the host inflammatory response [4]. Non-surgical removal of the microbial deposits, the mechanical root debridement by scaling and root planing (SRP) is considered as a standard of cause-related periodontal therapy. There is a substantial evidence that during supportive periodontal therapy, the progression of periodontitis can be controlled thorough mechanical plaque removal performed by the patient and the therapist eventually in conjunction with the use of antimicrobials [5]. The application of different antimicrobials adjunctive the mechanical removal of deposits by SRP has been tested. As antimicrobials, chlorhexidine, azithromycin, metronidazole, doxycycline, minocycline and tetracycline were used [6]. Results from a systematic review have indicated that the subgingival application of tetracycline fibres and of sustained released doxycycline and minocycline demonstrated significant benefit in clinical outcome (probing depth reduction) [6].

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An alternative not tested yet in periodontal therapy might be taurolidine. The antimicrobial activity of taurolidine has been known for about 20 years. Taurolidine is described as an unstable molecule in aqueous solution, masked methanol molecules are released which inactivate endotoxin [7]. Further derivatives are responsible for the antimicrobial action such as interaction with peptidoglycan [8]. First it was used in prevention and treatment of peritonitis [9]. The substance may be also for interest in dentistry. Recently it has been shown in an ex vivo-model that 2% taurolidine is effective in killing supragingival plaque [10]. In an in vitro-study, we compared a commercially available 2% taurolidine solution with a 0.1% chlorhexidine digluconate solution [11]. The minimal inhibitory concentrations (MICs) of taurolidine were all below 1 mg/ml of taurolidine (equivalent to 5% of the normally used concentration of that substance) with the exception of Candida albicans. This confirms an earlier study which determined MIC values against seven oral plaque species, among them one Fusobacterium nucleatum and one Prevotella intermedia strain [12]. Further our results clearly indicated, that taurolidine was active in a serum rich environment. Contrary the activity of chlorhexidine was dramatically decreased [11]. These findings are important, because gingival crevicular fluid contains up to 35% of the albumin found in serum [13]. The purpose of this in-vitro-study was to determine the antimicrobial activity of taurolidine in comparison with minocycline on microbial species associated with periodontitis within a mixed planktonic population and within a biofilm. The hypothesis was that taurolidine acts as antimicrobial as minocycline on a mixed microbiota associated with periodontitis. The antimicrobial activity was to be proven in killing assays, after exposure of the antimicrobials as well as within a biofilm. 2. Material and methods 2.1. Substances Test substances were taurolidine in a 2% w/v solution (Geistlich TauroSept®, Geistlich Pharma AG, Wolhusen, Switzerland) and minocycline (SigmaeAldrich, St. Louis, MO, USA). After determination of MICs and minimal bactericidal concentrations (MBCs) the final concentrations in the assays were 2.5, 5 and 10 mg/ml for taurolidine and 64, 128 and 256 mg/ml minocycline. Distilled water was used as a negative control. The tested concentrations were chosen in accordance with determined MBC values. 2.2. Microorganisms The following bacterial strains were tested as single bacterial species: A. actinomycetemcomitans Y4, P. gingivalis ATCC 33277, F. nucleatum ATCC 25586 and Streptococcus gordonii ATCC 10558. The mixed microbiota consisted of the following bacterial strains: S. gordonii ATCC 10558, Actinomyces naeslundii ATCC 12104, F. nucleatum ATCC 25586, Campylobacter rectus ATCC 33238, Eubacterium nodatum ATCC 33099, Eikenella corrodens ATCC 23834, P. intermedia ATCC 25611, Parvimonas micra ATCC 33270, P. gingivalis ATCC 33277, T. forsythia ATCC 43037, T. denticola ATCC 35405 and A. actinomycetemcomitans Y4. Before an experiment, all strains (except for T. denticola ATCC 35405) were precultivated on Schaedler agar plates (Oxoid, Basingstoke, UK) with 5% sheep blood in an anaerobic atmosphere or with 5% CO2 (A. actinomycetemcomitans Y4 and S. gordonii ATCC 10558). T. denticola was maintained in modified mycoplasma broth (BD, Franklin Lake, NJ) added by 1 mg/ml glucose, 400 mg/ml niacinamide, 150 mg/ml spermine tetrahydrochloride, 20 mg/ml Na

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isobutyrate enriched with 1 g/ml cysteine and 5 mg/ml cocarboxylase in anaerobic conditions. 2.3. Determination of the minimal inhibitory concentrations and minimal bactericidal concentrations First, the MICs of taurolidine and minocycline against the four single selected species as well as against the mixed population were determined. Taurolidine was tested in a two-fold dilution series starting from 10 mg/ml and minocycline from 256 mg/ml (final concentrations) with microbroth-dilution technique by using 96-well-microtiter plates. The test medium was double-concentrated Wilkins Chalgren broth (Oxoid) by adding 1:1 of the antimicrobial in an aqueous solution. MIC was determined as the lowest concentration without visible turbidity of the broth. The MBC was the lowest concentration without any growth of the subcultivations on the agar plates (equivalent to a reduction by 99.9% of the initial inoculum). 2.4. Killing A defined inoculum of microorganisms (5  106) was prepared in doubled concentrated nutrient media (Wilkins Chalgren broth). The test substances were added in a ratio 1:1. After 15 min, 30 min, 1 h, 2 h, 6 h as well as 24 h of incubation, the numbers of viable bacteria were determined by enumeration of colony forming units (cfu). (In case of the mixed microbiota only the total numbers of cfu were counted.) 2.5. Substantivity (growth inhibition after exposure) Microorganisms (108/ml) were exposed to antimicrobials in 3 ml of Wilkins-Chalgren broth for 2 h. After that, the suspensions were centrifuged 10 min at 5000 g. The supernatant was removed and 5 ml of nutrient broth (Wilkins Chalgren broth) was added. The cfu counts were determined after 1 h, 2 h, 4 h, 6 h and 24 h. All experiments were be made in independent replicates. 2.6. Activity against bacteria within biofilms In these experiments A. actinomycetemcomitans Y4, P. gingivalis ATCC 33277 were used to form a single species biofilm. In addition, a multispecies biofilm consisting of the 12 species was established. First the wells of 24-well-plates were covered with 100 ml of 25% v/ v inactivated human serum/well for 1 h. Then 1 ml of bacterial suspension was added. The medium was braineheart-infusionbroth (Oxoid Ltd.) with 5% blood (and 5 mg/ml cocarboxylase for the mixed population). The 24-well-plates were incubated in the appropriate atmosphere. After 60 h the medium was carefully exchanged. In case of the mixed biofilm P. gingivalis ATCC 33277, T. forsythia ATCC 43037 and T. denticola ATCC 35405 were again added to the nutrient medium before application to the wells. The renewed addition of selected bacterial strains guaranteed a sufficient number of these species within the biofilms [14]. After an additional incubation for 48 h, the medium was removed carefully, 100 ml of the antibiotic dilutions mixed with 100 ml of doubled concentrated Wilkins Chalgren broth were added. After 1 h, 800 ml of Wilkins Chalgren broth supplemented with 5% sheep blood (and 5 mg/ml cocarboxylase for the mixed population) was added. This simulates in part in vivo conditions, where a diluting effect of subgingivally applied antimicrobials can be assumed. The plates were incubated in the appropriate atmosphere overnight (18 h). Then, the medium was removed. Finally, the biofilm was carefully scraped, mixed by pipetting and cfu were enumerated after serial dilutions, spreading of each 25 ml on agar

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plates and incubation for 48 h (A. actinomycetemcomitans Y4) and 7 d (P. gingivalis ATCC 33277 and mixed microbiota). SEM photographs were taken to visualize the results. Exemplarily each one concentration of taurolidine (10 mg/ml) as well as one of minocycline (256 mg/ml) was chosen. Only the mixed microbiota was used. Samples were fixed in 2% glutaraldehyde in cacodylate buffer for 30 min, washed twice with cacodylate buffer and dehydrated using a graded ethanol series (10 min each concentration). Following critical point drying, samples were sputtercoated with gold and examined with a ZEISS LEO-1530 Gemini (Carl Zeiss NTS GmbH) equipped with a field emission electron gun at 10 keV. Moreover the potential inhibition of biofilm formation by the antimicrobials was tested. For that purpose wells were covered with 100 ml of the antimicrobial solution first. Thereafter, the biofilm was formed as described above for 24 h and 48 h. At these time-points the cfu counts were determined. These experiments were made in two independent experiments each in triplicates at least. 3. Results 3.1. Minimal inhibitory and minimal bactericidal concentrations The MICs of taurolidine against the four single species were always 0.31 mg/ml, the MBCs were one step higher with being 0.64 mg/ml. The used mixed microbiota was much less sensitive to taurolidine, MIC and the MBC was 2.5 mg/ml thus being 12.5% of the commercially available concentration of 2% w/v. Sensitivity of the included stains varied against minocycline, the highest MBC of 128 mg/ml was determined against the bacterial mixture (Table 1).

procedure should show a possible depot activity by the antimicrobials. In case of P. gingivalis ATCC 33277, these experiments confirm first the killing experiments with a complete killing after 2 h with no regrowth later. In general the substantivity of taurolidine seemed to be higher than those of minocycline resulting in lower cfu counts over the time (Suppl. Fig. 6). None of the used antimicrobials was able to kill completely bacteria within a mixture after an exposure of 2 h. Nevertheless, taurolidine reduced cfu counts in a concentration dependent manner. After 2 h of exposure to minocycline cfu counts were reduced with no clear influence by the used concentration (Fig. 1B). 3.4. Activity of antimicrobials on single-species biofilms The activity of the antimicrobials was tested on 4.5 d old biofilms. Even in the single-species biofilms, never a complete elimination was found. Taurolidine acted concentration dependent with the lowest cfu counts after application of 10 mg/ml taurolidine, here the reduction was 5 log10 cfu (P. gingivalis ATCC 33277) and 7 log10 cfu (A. actinomycetemcomitans Y4). Minocycline decreased the cfu counts by 1e2 log10 independent of the used concentration. In contrast to the results with an established biofilm, the biofilm formation of P. gingivalis was more sensitive to an antimicrobial influence than those of A. actinomycetemcomitans Y4. Coating the surface with 10 mg/ml taurolidine or 256 mg/ml minocycline prevented completely biofilm formation. In case of A. actinomycetemcomitans Y4 a reduced biofilm formation was found up to 48 h after coating the surface with 10 mg/ml taurolidine, 256 mg/ml minocycline reduced the biofilm formation only up to 24 h (Fig. 2). 3.5. Activity of antimicrobials on multi-species biofilms

3.2. Killing of planktonic bacteria Except for P. gingivalis ATCC 33277, the strains were completely killed by the highest used concentrations of 10 mg/ml taurolidine after 6 h and of 256 mg/ml minocycline after 24 h. In case of P. gingivalis ATCC 33277 the respective times were 2 h and 6 h suggesting a higher sensitivity of that strain to the antimicrobial killing. Most a concentration dependent activity was visible. Taurolidine in all used concentrations killed all bacteria up to 24 h; minocycline in the two lower tested concentrations did not eradicate F. nucleatum ATCC 25586 (Suppl. Fig. 5). Focussing on the mixture of 12 selected bacterial species associated with periodontitis, taurolidine was active in all three used concentrations to kill all bacteria, 256 mg/ml of minocycline reduced the bacterial counts by 5 log10 cfu (Fig. 1A). 3.3. Post-antimicrobial (depot) activity Bacteria were exposed to antibacterials for 2 h, thereafter the media were replaced by a medium free of antimicrobial agents. This

Table 1 MIC and MBC values of taurolidine and minocycline against selected oral species and a bacterial mixture consisting of 12 different species associated with periodontitis. Taurolidine (mg/ ml)

S. gordonii ATCC 10558 A. actinomycetemcomitans Y4 P. gingivalis ATCC 33277 F. nucleatum ATCC 25586 12 Species (mixed)

Minocycline (mg/ ml)

MIC

MBC

MIC

MBC

0.31 0.31 0.31 0.31 2.5

0.63 0.63 0.63 0.63 2.5

16 1 0.25 8 1

64 4 16 32 128

At the time of exposure to the antimicrobials established biofilms contained a mixture of bacteria ranged from 4.47 log10 cfu for A. actinomycetemcomitans Y4 until 7.28 log10 cfu for P. gingivalis ATCC 33277. The reduction of the cfu counts in the 4.5 d old multispecies biofilms was about 3 log10 cfu after application of any minocycline concentration and after using 10 mg/ml taurolidine. The activity of taurolidine was concentration-dependent with no activity by 2.5 mg/ml taurolidine. Multispecies biofilms formed for 24 h and 48 h contained bacteria in the range of 5.06 log10 cfu for F. nucleatum ATCC 25586 and 6.75 log10 cfu for P. micra ATCC 33270. Both 10 mg/ml taurolidine and 256 mg/ml minocycline inhibited multi-species biofilm formation but they were not able to prevent completely the adhesion of bacterial species (Fig. 3). SEM photographs were taken of multi-species biofilms without and after exposure to the antimicrobials. Photographs without exposure to antimicrobials confirm a thick biofilm composed of different bacteria. After exposure to the antimicrobials still a relative thick biofilm is visible. Photographs show bacteria with damaged membranes. In contrast to taurolidine not all shapes of bacteria seemed to be affected by minocycline (Fig. 4). 4. Discussion In this in vitro study the antibacterial activity of taurolidine was compared with that of minocycline an established topical antibacterial agent. The benefit of adjunctive administration of minocycline to nonsurgical periodontal therapy was reported in several clinical studies. Applied as a 2% gel it provided better outcome in terms of attachment gain and reduction of bleeding on probing than mechanical debridement alone [15]. Subgingival minocycline microspheres improve clinical parameters as well as they reduce

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Fig. 1. Numbers of viable bacteria (determined by colony forming units counts) of a mixed population consisting of 12 different species associated with periodontitis after continuous exposure to taurolidine and minocycline (A; killing) and 2 h of exposure to taurolidine and minocycline and subsequent cultivation in antimicrobial free media (B; postantimicrobial activity).

Fig. 2. Activity of taurolidine and minocycline on single-species biofilms of Aggregatibacter actinomycetemcomitans Y4 (A, C) and Porphyromonas gingivalis ATCC 33277 (B, D) A, B: On 4.5 d old biofilms taurolidine in concentrations of 0.25%, 0.5% and 1% and minocycline in concentration of 64 mg/ml, 128 mg/ml and 256 mg/ml were added for 1 h and thereafter diluted 1:4 for 18 h, before cfu counts were determined. C, D: After coating surface with 1% of taurolidine and 256 mg/ml of minocycline, biofilms were formed for 24 h and 48 h.

bacteria associated with periodontitis when compared with SRP alone [16]. In initial peri-implantitis, its adjunctive use to mechanical treatment has shown reductions in probing depths for 12 months [17]. MIC values of taurolidine were low against single tested species. They went up to 12.5% of the commercially available concentration when a mixture of 12 oral species was tested. Taurolidine in all tested concentrations (down to 2.5 mg/ml) was able to kill all included single bacterial species as well as the mixture of 12 oral species within 24 h. Minocycline in the tested concentrations did not completely eliminate the bacterial mixture in any of the tested concentrations (up to 256 mg/ml). Mode of action is different

between the two tested substances. Minocycline as doxycycline or tetracycline inhibits protein synthesis [18]. Taurolidine interferes with bacterial cell wall components [7,8] which might directly kill bacteria. Substantivity is needed for topical applications of antimicrobials because a fast diluting activity can be expected. E.g., gingival fluid in periodontitis patients has a volume of up to 1.5 ml and a turnover of up to 44 ml/h [19]. The used method was adapted to protocols determining a postantibiotic effect [20] and the in vivo-situation. Bacteria have been exposed to the antimicrobials for 2 h. Later the media were replaced instead of making a dilution only. The used concentrations were equal for all included microorganisms. They

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Fig. 3. Activity of taurolidine and minocycline on multi-species biofilms consisting of 12 different species associated with periodontitis A: On 4.5 d old biofilms taurolidine in concentrations of 0.25%, 0.5% and 1% and minocycline in concentration of 64 mg/ml, 128 mg/ml and 256 mg/ml were added for 1 h and thereafter diluted 1:4 for 18 h, before cfu counts were determined. B: After coating surface with 1% of taurolidine and 256 mg/ml of minocycline, biofilms were formed for 24 h and 48 h.

were in the range of 8-folde32-fold MIC for single species and 1fold e 4-fold MIC for the mixture when testing taurolidine. For minocycline the values were much higher, the range was between 4-fold up to 1000-fold MIC. Although the relation to the MIC was in general higher for minocycline than for taurolidine, the “postantibacterial” activity was more remarkable on this compound when comparing with minocycline. Subgingival plaque represents a biofilm consisting of hundreds of different taxa [21], communication and transfer of DNA occurs between the members of a biofilm [22]. In biofilms limited growth, protein synthesis, metabolic activity and an increased mutation frequency of bacteria as well as a polymer matrix around microcolonies contribute to lower susceptibility to antibacterials in comparison with planktonic bacteria [23]. Recently a study

determined the activity of antibacterials in concentrations found after systemic application in gingival crevicular fluid on a multispecies biofilm in vitro; a remarkable reduction of the bacterial counts was not found [24]. Here, the activity of topically applied formulations was simulated. Single- and multi-species biofilms were studied. Taurolidine reduced the cfu counts of an existing single-species biofilm clearly. The activity was reduced on a multispecies biofilm. Only the highest tested concentration was able to reduce the cfu counts by up to 3 log10. The reduction was similar for minocycline in all tested concentrations with no clear difference between single- and multi-species biofilm. Taurolidine clearly inhibited the biofilm formation up to 48 h whereas minocycline had a reducing activity on the multi-species biofilm only up to 24 h. In another study, minocycline exerted

Fig. 4. SEM photographs of multi-species biofilms of 12 different species associated with periodontitis formed for 4.5 d and thereafter without (A, B) and with exposure to 10 mg/ml taurolidine for 1 h followed by 2 mg/ml taurolidine (one fifth of the first concentration) for 24 h (C) and 256 mg/ml minocycline for 1 h followed by 51.2 mg/ml minocycline (one-fifth of the first concentration) for 24 h (D).

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only an activity on an early A. actinomycetemcomitans biofilm but not when applied to a biofilm formed already for 24 h [25]. In 10 d old P. gingivalis biofilms viability decreased after permanent exposure to minocycline for 24 h up to 144 h but to less extent when compared with chlorhexidine [26]. The inhibiting activity of taurolidine on biofilm formation in catheters has been evaluated in in vitro studies and clinical trials. In vitro exposure to taurolidinecitrate resulted in a reduction by five log10 cfu for major pathogens of catheter-associated infections [27]. Taurolidine 2%/polyvinylpyrolidine was in vitro more active against pathogens inoculated in catheters [28]. In vivo, taurolidine reduced the numbers of bloodstream infections in comparison with the standard heparin [29]. In chronic patients on haemodialysis locking permanent catheters with a taurolidineecitrateeheparin filling eradicated pathogens [30]. In all assays a clear concentration dependent activity of taurolidine was found. A concentration closed to those in commercial products cannot be expected for a longer time in the periodontal region. The high substantivity of taurolidine is advantageous. But nevertheless usage of sustained or controlled released devices might be essential. Minocycline has been incorporated in such devices. It was used as a 2% gel [15]. Better known are the microspheres, which had been proven to effective in several clinical trials [16]. Concentrations for most experiments were chosen according to MBC values. Thus the concentrations of taurolidine were higher than those of minocycline. Our results suggest using a higher concentration than 2% w/v taurolidine in a sustained or controlled released device. Taken together, the present data indicate that taurolidine is active against species associated with periodontitis, even within biofilms. Nevertheless, a complete elimination of complex biofilms by taurolidine alone seems to be impossible and underlines the importance of mechanical removal of biofilms prior to application of taurolidine. Taurolidine in a sustained or controlled-release device may have potential as an adjunctive antimicrobial treatment in periodontitis and should be further evaluated in vitro and in clinical trials. Acknowledgement The present study was funded by Geistlich Pharma AG (UB13194), Wolhusen, Switzerland. The authors declare that they have no conflicts of interest. The authors would like to thank Marianne Weibel (Department of Periodontology, Laboratory of Oral Microbiology, School of Dental Medicine, University of Bern) for technical assistance. Appendix A. Supplementary data Supplementary data related to this article can be found at http:// dx.doi.org/10.1016/j.anaerobe.2014.11.008. References [1] Darveau RP. Periodontitis: a polymicrobial disruption of host homeostasis. Nat Rev Microbiol 2010;8:481e90. [2] Teles R, Teles F, Frias-Lopez J, Paster B, Haffajee A. Lessons learned and unlearned in periodontal microbiology. Periodontol 2000 2013;62:95e162. [3] Hajishengallis G, Darveau RP, Curtis MA. The keystone-pathogen hypothesis. Nat Rev Microbiol 2012;10:717e25.

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[4] Johansson A. Aggregatibacter actinomycetemcomitans leukotoxin: a powerful tool with capacity to cause imbalance in the host inflammatory response. Toxins (Basel) 2011;3:242e59. [5] Hancock EB, Newell DH. Preventive strategies and supportive treatment. Periodontol 2000 2001;25:59e76. [6] Matesanz-Perez P, Garcia-Gargallo M, Figuero E, Bascones-Martinez A, Sanz M, Herrera D. A systematic review on the effects of local antimicrobials as adjuncts to subgingival debridement, compared with subgingival debridement alone, in the treatment of chronic periodontitis. J Clin Periodontol 2013;40: 227e41. [7] Gidley MJ, Sanders JK, Myers ER, Allwood MC. The mode of antibacterial action of some ‘masked’ formaldehyde compounds. FEBS Lett 1981;127:225e7. [8] Caruso F, Darnowski JW, Opazo C, Goldberg A, Kishore N, Agoston ES, et al. Taurolidine antiadhesive properties on interaction with E. coli; its transformation in biological environment and interaction with bacteria cell wall. 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In-vitro activity of taurolidine on single species and a multispecies population associated with periodontitis.

The antimicrobial activity of taurolidine was compared with minocycline against microbial species associated with periodontitis (four single strains a...
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