Papers by Arie Van Winkelhoff
mBio, Jan 30, 2018
The keystone oral pathogen is associated with severe periodontitis. Intriguingly, this bacterium ... more The keystone oral pathogen is associated with severe periodontitis. Intriguingly, this bacterium is known to secrete large amounts of an enzyme that converts peptidylarginine into citrulline residues. The present study was aimed at identifying possible functions of this citrullinating enzyme, named peptidylarginine deiminase (PPAD), in the periodontal environment. The results show that PPAD is detectable in the gingiva of patients with periodontitis, and that it literally neutralizes human innate immune defenses at three distinct levels, namely bacterial phagocytosis, capture in neutrophil extracellular traps (NETs), and killing by the lysozyme-derived cationic antimicrobial peptide LP9. As shown by mass spectrometry, exposure of neutrophils to PPAD-proficient bacteria reduces the levels of neutrophil proteins involved in phagocytosis and the bactericidal histone H2. Further, PPAD is shown to citrullinate the histone H3, thereby facilitating the bacterial escape from NETs. Last, PPA...
Journal of proteome research, Jan 7, 2016
Porphyromonas gingivalis is an oral pathogen associated with the inflammatory disease periodontit... more Porphyromonas gingivalis is an oral pathogen associated with the inflammatory disease periodontitis. Periodontitis and P. gingivalis¬ have been associated with rheumatoid arthritis. One of the hallmarks of rheumatoid arthritis is the loss of tolerance against citrullinated proteins. Citrullination is a post-translational modification of arginine residues, leading to a change in structure and function of the respective protein. This modification, which is catalysed by peptidylarginine deiminases (PAD), plays a role in several physiological processes in the human body. Interestingly, P. gingivalis secretes a citrullinating enzyme, known as P. gingivalis PAD (PPAD), which targets bacterial and human proteins. Since the extent of P. gingivalis protein citrullination by PPAD was not yet known, the present study was aimed at identifying the extracellular proteome and citrullinome of P. gingivalis. To this end, extracellular proteins of two reference strains, two PPAD-deficient mutants and...
Journal of Periodontology, 2015
Background: Currently, in the field of rheumatology, there is much attention given towards the po... more Background: Currently, in the field of rheumatology, there is much attention given towards the possible causality between periodontitis and rheumatoid arthritis (RA), specifically regarding the role of Porphyromonas gingivalis (Pg). This bacterium is unique, having a citrullinating enzyme. Antibodies against citrullinated proteins are rather specific for RA.Methods: Because causality is ultimately tested in longitudinal cohort studies which currently do not exist for periodontitis and RA, this commentary applied Bradford Hill criteria on the existing literature to assess causality as the most likely interpretation of this association.Conclusions: From an epidemiologic point of view, patients with RA have a higher incidence of periodontal disease than individuals without RA. In addition, there is a dose‐response pattern in the association between the severity of periodontitis and RA disease activity. There are indications that periodontitis precedes RA, but there is no evidence yet a...
Arthritis Research & Therapy, 2012
Nederlands Tijdschrift voor Tandheelkunde, 2012
In 1986 stond de orale microbiologie, vooral de microbiologie van parodontale infecties, volop in... more In 1986 stond de orale microbiologie, vooral de microbiologie van parodontale infecties, volop in de belangstelling. De parodontologie was enorm in opkomst en daarmee ook de ontwikkeling van de microbiologie en de immunologie van infectieuze aandoeningen van het parodontium. Op nationale en internationale podia werd gediscussieerd over de nieuwe bacterie Actinobacillus actinomycetemcomitans en over de rol van antibiotica bij de behandeling van ernstige parodontitis. Sinds de jaren '80 van de vorige eeuw is er veel veranderd in de parodontologie, niet in de laatste plaats doordat een nieuwe behandelwijze zijn definitieve intrede deed: de orale implantologie. Daarmee ontstond een nieuwe ziekte: peri-implantaire infectie.
Clinical Oral Investigations, 2016
Objectives The aim of the present study was to compare the composition of the periodontal microfl... more Objectives The aim of the present study was to compare the composition of the periodontal microflora at baseline (T0) with the submucosal microflora at least 1 year after implant placement (T1) in periodontally healthy patients. Material and methods For all 169 consecutive patients that visited our clinic during 1 year, we determined their periodontal parameters, implant mucosal index, and presence of implant calculus. At T0, self-reported smoking status was recorded and subgingival and submucosal biofilm samples were obtained and analyzed for the presence and numbers of selected periodontal pathogens. All measurements were repeated at T1. Results One hundred twenty patients completed the study. Periodontal parameters were stable or had improved at T1. The total bacterial load was lower at implant sites (P < 0.05). The prevalence of Porphyromonas gingivalis was low at baseline, but at T1, detection rate and numbers were higher at implant sites compared to dentate sites. At T1, the frequency of detection of P. gingivalis (P = 0.01), Parvimonas micra (P = 0.018), and Fusobacterium nucleatum (P = 0.035) was higher in smoking patients (n = 23) than in non-smokers (n = 97). Conclusions Colonization of the submucosal peri-implant area is similar to the composition of subgingival microbiota. Smoking has a measurable effect on the colonization of implant-associated biofilms and may select for P. gingivalis, P. micra, and F. nucleatum. Clinical relevance The colonization of implants by wellknown periodontal pathogens is very similar to that in normal dentition, also in a healthy cohort. Smoking status was related with the prevalence of periodontal pathogens where smokers harbored more often periodontal pathogens such as P. gingivalis, P. micra, and F. nucleatum.
Journal of Clinical Periodontology, 2013
Background: There has been an explosion in research into possible associations between periodonti... more Background: There has been an explosion in research into possible associations between periodontitis and various systemic diseases and conditions. Aim: To review the evidence for associations between periodontitis and various systemic diseases and conditions, including chronic obstructive pulmonary disease (COPD), pneumonia, chronic kidney disease, rheumatoid arthritis, cognitive impairment, obesity, metabolic syndrome and cancer, and to document headline discussions of the state of each field. Periodontal associations with diabetes, cardiovascular disease and adverse pregnancy outcomes were not discussed by working group 4. Results: Working group 4 recognized that the studies performed to date were largely cross-sectional or case-control with few prospective cohort studies and no randomized clinical trials. The best current evidence suggests that periodontitis is characterized by both infection and pro-inflammatory events, which variously manifest within the systemic diseases and disorders discussed. Diseases with at least minimal evidence of an association with periodontitis include COPD, pneumonia, chronic kidney disease, rheumatoid arthritis, cognitive impairment, obesity, metabolic syndrome and cancer. The working group agreed that there is insufficient evidence to date to infer causal relationships with the exception that organisms originating in the oral microbiome can cause lung infections. Conclusions: The group was unanimous in their opinion that the reported associations do not imply causality, and establishment of causality will require new studies that fulfil the Bradford Hill or equivalent criteria. Precise and community-agreed case definitions of periodontal disease states must be implemented systematically to enable consistent and clearer interpretations of studies of the relationship to systemic diseases. The members of the working group were unanimous in their opinion that to develop data that best inform clinicians, investigators and the public, studies should focus on robust disease outcomes and avoid surrogate endpoints. It was concluded that because of the relative immaturity of the body of evidence for each of the purported relationships, the field is wide open and the gaps in knowledge are large.
FEMS Immunology …, 2005
Periodontitis is a multi-factorial chronic inflammatory and destructive disease of the tooth-supp... more Periodontitis is a multi-factorial chronic inflammatory and destructive disease of the tooth-supporting tissues. Quantitative anaerobic culture techniques have been used for microbial diagnosis of the different forms of the disease. The aim of this study was to compare real-time PCR with quantitative anaerobic culture for detection and quantification of 5 prominent periodontal pathogens. Real-time PCR assays with the 16s rRNA genes of Actinobacillus actinomycetemcomitans, Prevotella intermedia, Tannerella forsythensis, Peptostreptococcus micros and Fusobacterium spp. were developed. The PCR was validated on pure cultures of various bacterial strains. Subsequently, subgingival plaque samples from 259 adult patients with periodontitis were analyzed with quantitative anaerobic culture and real-time PCR. A standard curve for DNA quantification was created for each primer-probe set based on colony-forming units equivalents. All bacterial species were correctly identified. The lower limits of detection by PCR varied between 1-50 colony-forming units equivalents depending on the species. No cross-reactivities with heterologous DNA of other bacterial species were observed. Real-time PCR results showed a high degree of agreement with anaerobic culture results. Real-time PCR is a reliable alternative for diagnostic quantitative anaerobic culture of subgingival plaque samples.
Arthritis Research & Therapy, 2015
Journal of Periodontology, 2015
Currently, in the field of rheumatology, there is much attention given towards the possible causa... more Currently, in the field of rheumatology, there is much attention given towards the possible causality between periodontitis and rheumatoid arthritis (RA), specifically regarding the role of Porphyromonas gingivalis (Pg). This bacterium is unique, having a citrullinating enzyme. Antibodies against citrullinated proteins are rather specific for RA. Because causality is ultimately tested in longitudinal cohort studies which currently do not exist for periodontitis and RA, this commentary applied Bradford Hill criteria on the existing literature to assess causality as the most likely interpretation of this association. From an epidemiologic point of view, patients with RA have a higher incidence of periodontal disease than individuals without RA. In addition, there is a dose-response pattern in the association between the severity of periodontitis and RA disease activity. There are indications that periodontitis precedes RA, but there is no evidence yet available to show that Pg plays a direct role in this temporal relationship. The role of the unique characteristic of citrullination by Pg remains unexplained. However, in animal models, citrullination by Pg plays a distinct role in the development and aggravation of experimental arthritis. Although the role of Pg in RA remains speculative, a causative role for periodontitis as a chronic inflammatory disease caused by infectious agents in RA seems biologically plausible.
Annals of the Rheumatic Diseases
Journal of Clinical Periodontology, 2014
The aim of the study was to evaluate the effect of full-mouth tooth extraction on the oral microf... more The aim of the study was to evaluate the effect of full-mouth tooth extraction on the oral microflora, with emphasis on the presence and load of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Adult patients (n = 30), with moderate to advanced periodontitis and scheduled for full-mouth tooth extraction, were consecutively selected. Prior to and 1 and 3 months after full-mouth tooth extraction saliva, tongue, buccal and gingival mucosa and subgingival plaque/prosthesis samples were obtained. Aerobic and anaerobic culture techniques and quantitative real-time polymerase chain reaction (qPCR) were employed for the detection of oral pathogens. Full-mouth tooth extraction resulted in reduction below detection level of A. actinomycetemcomitans and P. gingivalis in 15 of 16 and 8 of 16 previously positive patients using culture techniques and qPCR, respectively. Those patients remaining qPCR positive showed a significant reduction in load of these bacteria. Full-mouth tooth extraction significantly changes the oral microflora. These changes include reduction of A. actinomycetemcomitans and P. gingivalis, frequently to levels below detection threshold. In some patients, A. actinomycetemcomitans and P. gingivalis can persist in the edentulous oral cavity up to 3 months after full-mouth tooth extraction.
Odontology, 2014
This study assessed the subgingival occurrence of the flagellated, Gram-negative, anaerobic rod C... more This study assessed the subgingival occurrence of the flagellated, Gram-negative, anaerobic rod Centipeda periodontii in chronic periodontitis and periodontal health/gingivitis with species-specific nucleic acid probes, and evaluated the in vitro resistance of subgingival isolates to therapeutic levels of amoxicillin, metronidazole, and doxycycline. Subgingival plaque biofilm specimens from 307 adults with chronic periodontitis, and 48 adults with periodontal health/localized gingivitis, were evaluated with digoxigenin-labeled, whole-chromosomal, DNA probes to C. periodontii ATCC 35019 possessing a 10(4) cell detection threshold. Fifty-two C. periodontii subgingival culture isolates were assessed on antibiotic-supplemented enriched Brucella blood agar for in vitro resistance to either amoxicillin at 2 µg/ml, metronidazole at 4 µg/ml, or doxycycline at 2 µg/ml. A significantly greater subgingival occurrence of C. periodontii was found in chronic periodontitis subjects as compared to individuals with periodontal health/gingivitis (13.4 vs. 0 %, P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.003), although high subgingival counts of the organism (≥10(6) cells) were rarely detected (1.3 % of chronic periodontitis subjects). In vitro resistance was not found to amoxicillin or metronidazole, and to doxycycline in only 2 (3.9 %) of the 52 C. periodontii clinical isolates studied. These findings indicate that C. periodontii is not a major constituent of the subgingival microbiome in chronic periodontitis or periodontal health/gingivitis. The potential contribution of C. periodontii to periodontal breakdown in the few chronic periodontitis subjects who yielded high subgingival levels of the organism remains to be delineated. C. periodontii clinical isolates were susceptible in vitro to therapeutic concentrations of three antibiotics frequently used in treatment of human periodontitis.
Periodontology 2000, 2005
Oral Microbiology and Immunology, 2007
Journal of Clinical Periodontology, 2011
Slot DE, Kranendonk AA, Van der Reijden WA, Van Winkelhoff AJ, Rosema NAM, Schulein WH, Van der V... more Slot DE, Kranendonk AA, Van der Reijden WA, Van Winkelhoff AJ, Rosema NAM, Schulein WH, Van der Velden U, Van der Weijden FA. Adjunctive effect of a watercooled Nd:YAG laser in the treatment of chronic periodontitis.
Journal of Clinical Periodontology, 2002
Heydenrijk K, Raghoebar GM, Meijer HJA, van der Reijden WA, van Winkelhoff A-J, Stegenga B. Two-p... more Heydenrijk K, Raghoebar GM, Meijer HJA, van der Reijden WA, van Winkelhoff A-J, Stegenga B. Two-part implants inserted in a one-stage or a two-stage procedure. A prospective comparative study. J Clin Periodontol 2002; 29: 900-909.
Journal of Clinical Periodontology, 2007
Journal of Clinical Periodontology, 2002
putative periodontal pathogens in subjects with and without periodontal destruction van Winkelhof... more putative periodontal pathogens in subjects with and without periodontal destruction van Winkelhoff AJ, Loos BG, van der Reijden WA, van der Velden U. Porphyromonas gingivalis, Bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction.
Infection, Genetics and Evolution, 2010
The periodontal pathogen Aggregatibacter actinomycetemcomitans that comprises six serotypes (a-f)... more The periodontal pathogen Aggregatibacter actinomycetemcomitans that comprises six serotypes (a-f), is often identified by PCR-based techniques targeting the 16S rRNA gene. In this study, 16S rRNA gene sequence analysis revealed an aberrant cluster of 19 strains within serotype e, denoted as serotype e 0 . The 16S rRNA gene sequence similarities found between serotype e 0 strains ranged from 99.7% to 100.0%, whereas 96.8-97.5% sequence similarity was obtained with members of the other serotypes, indicating that the serotype e 0 strains might not be true members of A. actinomycetemcomitans. However, DNA-DNA hybridizations between a representative serotype e 0 strain and representative strains of serotypes b, d and e of A. actinomycetemcomitans revealed 68-75% DNA-DNA relatedness, demonstrating that the serotype e 0 strains do belong to the species A. actinomycetemcomitans. AFLP analysis of 33 A. actinomycetemcomitans strains, representing all serotypes (a-f), but mainly serotype e 0 strains, showed that the latter form a distinct cluster, demonstrating that these strains are also closely related on the whole genome level. Moreover, the serotype e 0 strains were unable to ferment starch and glycogen in contrast to almost all other A. actinomycetemcomitans strains tested. Overall, the data obtained in this study suggest that the serotype e 0 strains form an evolutionary relatively stable distinct subgroup within A. actinomycetemcomitans. ß
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Papers by Arie Van Winkelhoff