Papers by Lamprini Karygianni
Journal of Endodontics, 2014
The aim of the present study was to analyze the microbiota of primary and secondary/ persistent e... more The aim of the present study was to analyze the microbiota of primary and secondary/ persistent endodontic infections of patients undergoing endodontic treatment with respect to clinical and radiographic findings. Methods: Samples from the root canals of 21 German patients were taken using 3 sequential sterile paper points. In the case of a root canal filling, gutta-percha was removed with sterile files, and samples were taken using sterile paper points. The samples were plated, and microorganisms were then isolated and identified morphologically by biochemical analysis and sequencing the 16S rRNA genes of isolated microorganisms. Results: In 12 of 21 root canals, 33 different species could be isolated. Six (50%) of the cases with isolated microorganisms were primary, and 6 (50%) cases were endodontic infections associated with root-filled teeth. Twelve of the isolated species were facultative anaerobic and 21 obligate anaerobic. Monomicrobial infections were found for Enterococcus faecalis and Actinomyces viscosus. E. faecalis was most frequently isolated in secondary endodontic infections (33%). Moraxella osloensis was isolated from a secondary endodontic infection that had an insufficient root canal filling accompanied by a mild sensation of pain. A new bacterial composition compromising Atopobium rimae, Anaerococcus prevotii, Pseudoramibacter alactolyticus, Dialister invisus, and Fusobacterium nucleatum was recovered from teeth with chronic apical abscesses. Conclusions: New bacterial combinations were found and correlated to clinical and radiographic findings, particularly to chronic apical abscesses. M. osloensis was detected in root canals for the second time and only in German patients. (J Endod 2014;40:670-677)
Journal of medical microbiology, Jan 19, 2016
Bacterially-driven mucosal inflammation and the development of periimplantitis can lead to oral i... more Bacterially-driven mucosal inflammation and the development of periimplantitis can lead to oral implant failure. In this study, initial bacterial adhesion after 2 h and biofilm formation after 1 day and 3 days were analyzed in situ on novel 3 mol% yttria-stabilized tetragonal zirconia polycrystal samples (Zr; 3Y-TZP), as well as on alumina and niobium co-doped yttria-stabilized tetragonal zirconia samples (Al-Zr; Al2O3/Y(Nb)-TZP). Pure titanium implant material (Ti) and bovine enamel slabs (BES) served as controls. The initially adherent oral bacteria were determined by DAPI-staining. Biofilm thickness, surface covering grade and content of oral streptococci within the biofilm were measured by fluorescence in situ hybridization. No significant differences between the ceramic and titanium surfaces were detectable for either initial bacterial adhesion or the oral streptococci content of the in situ biofilm. The values of oral biofilm thickness on the implant surfaces were almost doubl...
Frontiers in Microbiology, 2016
Oral diseases such as caries and periodontitis are mainly caused by microbial biofilms. Antibioti... more Oral diseases such as caries and periodontitis are mainly caused by microbial biofilms. Antibiotic therapy has reached its limits with regard to antimicrobial resistance, and new therapeutic measures utilizing natural phytochemicals are currently a focus of research. Hence, this systematic review provides a critical presentation of the antimicrobial effects of various medicinal herbs against in vitro, ex vivo, and in situ formed multispecies oral biofilms. Searches were performed in three English databases (PubMed, EMBASE, CAMbase) and the electronic archives of five German journals from the times of their establishment until October 10th, 2014, with the search terms "(plant extracts OR herbal extracts OR plant OR herb) AND (oral biofilm OR dental biofilm OR dental plaque OR oral disease OR dental disease)." The pooled data were assessed according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines (PRISMA). Initially, 1848 articles were identified, out of which 585 full-text articles were screened, 149 articles were reevaluated for eligibility and finally, 14 articles met all inclusion criteria. The data of 14 reports disclosed enhanced antiadhesive and antibiofilm activity by the plant extracts obtained from Vitis vinifera, Pinus spp., Coffea canephora, Camellia sinensis, Vaccinium macrocarpon, Galla chinensis, Caesalpinia ferrea Martius, Psidium cattleianum, representative Brazilian plants and manuka honey. Overall, a positive correlation was revealed between herb-based therapies and elimination rates of all types of multispecies oral biofilms. In that context, integrating or even replacing conventional dental therapy protocols with herbal-inspired treatments can allow effective antimicrobial control of oral biofilms and thus, dental diseases.
Frontiers in Microbiology, 2016
Enterococci have gained significance as the cause of nosocomial infections; they occur as food co... more Enterococci have gained significance as the cause of nosocomial infections; they occur as food contaminants and have also been linked to dental diseases. E. faecalis has a great potential to spread virulence as well as antibiotic resistance genes via horizontal gene transfer. The integration of food-borne enterococci into the oral biofilm in-vivo has been observed. Therefore, we investigated the virulence determinants and antibiotic resistance of 97 E. faecalis isolates from the oral cavity, food, and clinical specimens. In addition, phenotypic expression of gelatinase and cytolysin were tested, in-vitro biofilm formation was quantified and isolates were compared for strain relatedness via pulsed field gel electrophoresis (PFGE). Each isolate was found to possess two or more virulence genes, most frequently gelE, efaA, and asa1. Notably, plaque/saliva isolates possessed the highest abundance of virulence genes, the highest levels of phenotypic gelatinase and hemolysin activity and concurrently a high ability to form biofilm. The presence of asa1 was associated with biofilm formation. The biofilm formation capacity of clinical and plaque/saliva isolates was considerably higher than that of food isolates and they also showed similar antibiotic resistance patterns. These results indicate that the oral cavity can constitute a reservoir for virulent E. faecalis strains possessing antibiotic resistance traits and at the same time distinct biofilm formation capabilities facilitating exchange of genetic material.
Clinical oral investigations, Jan 27, 2015
Biomaterial surfaces are at high risk for initial microbial colonization, persistence, and concom... more Biomaterial surfaces are at high risk for initial microbial colonization, persistence, and concomitant infection. The rationale of this study was to assess the initial adhesion on novel implant surfaces of Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans upon incubation. The tested samples were 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP) samples with nitrogen-doped hydrogenated amorphous carbon (a-C:H:N) coating (A) and 3Y-TZP samples coated with ceria-stabilized zirconia-based (Ce-TZP) composite and a-C:H:N (B). Uncoated 3Y-TZP samples (C) and bovine enamel slabs (BES) served as controls. Once the surface was characterized, the adherent microorganisms were quantified by estimating the colony-forming units (CFUs). Microbial vitality was assessed by live/dead staining, and microbial-biomaterial surface topography was visualized by scanning electron microscopy (SEM). Overall, A and B presented the lowest...
Objective: Bacterial adhesion to implant biomaterials leads to biofilm formation and hence, to in... more Objective: Bacterial adhesion to implant biomaterials leads to biofilm formation and hence, to infection and treatment failure. The aim of this study was to examine the initial microbial colonization on different implant materials in vitro. Method: Two novel implant biomaterials were incubated with Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans for 2 h: yttria-stabilized tetragonal zirconia polycrystal surfaces (Y-TZP) with nitrogen-containing hydrogenated amorphous carbon (a-C:H:N) coating (A) and Y-TZP surfaces with zirconia-based composite (Ce-TZP) and a-C:H:N coatings (B). Bovine enamel slabs (BES) served as control. The adherent microorganisms were quantified by determination of colony-forming units (CFUs) and visualized using scanning electron microscopy (SEM), DAPI and live/dead staining. Result: A and B showed comparable CFUs sheltering greater amounts of E. coli and lower numbers of E. faecalis, S. aureus and P. a...
Objectives: Bacterial colonization of enamel has been demonstrated to be crucial etiological fact... more Objectives: Bacterial colonization of enamel has been demonstrated to be crucial etiological factor in biofilm-mediated diseases such as caries, gingivitis and periodontal disease. The aim of the present in situ study was to establish a novel method for thorough visualization and quantification of initial bacterial adherence to enamel surfaces. Methods: For this purpose, bovine enamel slabs were fixed at buccal sites of individual upper jaw acrylic devices worn by a healthy volunteer for 3 and 5 days, respectively. Six disinfected enamel plates were placed in the interdental area between upper premolars and molars. Adherent bacteria on the slabs were visualized and quantified with fluorescence in situ hybridization (FISH) of eubacteria, Streptococcus spp. and Fusobacterium nucleatum using the ScanR screening platform. A statistical analysis of variance (ANOVA) evaluated the results. Results: The ScanR allowed visualization of randomly distributed bacteria on the complete enamel plat...
Photodiagnosis and photodynamic therapy, Jan 19, 2015
The aim of this study was to evaluate the effect of photodynamic therapy (PDT) on Enterococcus fa... more The aim of this study was to evaluate the effect of photodynamic therapy (PDT) on Enterococcus faecalis biofilms in artificially infected root canals using modified photosensitizers and passive ultrasonic activation. Two hundred and seventy extracted human teeth with one root canal were instrumented utilizing ProTaper files, autoclaved, infected with E. faecalis T9 for 72hours and divided into different groups: irrigation with 3% sodium hypochlorite (NaOCl), 20% ethylenediaminetetraacetic acid (EDTA), or 20% citric acid, PDT without irrigation, PDT accompanied by irrigation with NaOCl, EDTA, or citric acid, PDT using an EDTA-based photosensitizer or a citric-acid-based photosensitizer and PDT with ultrasonic activation of the photosensitizer. A 15mg/ml toluidine blue served as the photosensitizer, activated by a 100 mW LED light source. Sterile paper points were used for sampling the root canals and dentin chips were collected to assess the remaining contamination after treatment. S...
International endodontic journal, Jan 29, 2014
To evaluate whether artificial resin teeth could replace extracted human teeth in pre-clinical en... more To evaluate whether artificial resin teeth could replace extracted human teeth in pre-clinical endodontic training and if this teaching approach influences the outcome of root canal treatment on patients. In a pre-clinical training course, students of group 1 (n = 44) performed simulated endodontic exercises on four plastic blocks and three extracted human teeth. Students of group 2 (n = 45) performed their exercises on plastic blocks and artificial resin teeth (Real-T Endo, Acadental, Lenexa, KS, USA). Both groups performed their first root canal treatments on patients in the following term. Radiographs taken during root canal treatment were used for the evaluation of treatment outcome. Distances between the master cone or the root filling and the radiographic apex as well as iatrogenic errors were assessed, and comparisons were made using Fischer's exact test. In the pre-clinical course root canal treatments performed by students of group 2 were more often classified as accept...
Applied and Environmental Microbiology, 2014
BMC Research Notes, 2014
Background: Several studies have shown the antibacterial effectiveness of 0.2% chlorhexidine (CHX... more Background: Several studies have shown the antibacterial effectiveness of 0.2% chlorhexidine (CHX) in both in vitro and in vivo studies. In this way, CHX comes directly in contact with saliva. This in vitro study aimed at investigating the possible neutralizing effect of saliva on CHX. Methods: Saliva samples (12 ml) were collected from twenty healthy volunteers. The aerobic and anaerobic bacterial counts in saliva were determined on Colombia blood agar (CBA) and yeast cysteine agar (HCB), respectively. Saliva from each subject was divided among 4 experimental groups (3 ml/group). Samples were centrifuged at 4000 g for 10 min. The centrifuged salivary bacteria were incubated with the following solutions: 0.2% CHX in saliva, CHX in saliva with 7% ethanol, CHX in 0.9% NaCl, CHX in 0.9% NaCl with 7% ethanol. After exposure for 1 min or 3 min to these CHX solutions, the CHX was neutralized and the bacteria were cultivated, after which the number of colony forming units (aerobic and anaerobic) was determined. Results: CHX reduced the CFU in all groups significantly (p = 0.0001). Therefore, CHX had a similar effect on both aerobic and anaerobic microorganisms. Significantly more bacteria survived the effect of CHX when kept in salivary solution. This effect from saliva could be compensated by the addition of ethanol. In the absence of saliva there was no significant difference observed in the effectiveness of CHX with respect to ethanol. Prolonging the exposure time to 3 min enhanced the effectiveness of CHX. Conclusions: The effect of saliva on the antimicrobial activity of CHX was weak albeit statistically significant. However, addition of 7% ethanol compensates this effect. The impact of saliva on the reduction of the antimicrobial efficacy of mouthrinses such as CHX needs to be taken into consideration with regard to improving their antibacterial properties.
BMC Microbiology, 2014
Background: Lactobacillus represents a large genus with different implications for the human host... more Background: Lactobacillus represents a large genus with different implications for the human host. Specific lactobacilli are considered to maintain vaginal health and to protect from urogenital infection. The presence of Lactobacillus species in carious lesions on the other hand is associated with progressive caries. Despite their clinical significance, species-level identification of lactobacilli still poses difficulties and mostly involves a combination of different phenotypic and genotypic methods. This study evaluated rapid MALDI-TOF MS analysis of vaginal and oral Lactobacillus isolates in comparison to 16S rDNA analysis. Results: Both methods were used to analyze 77 vaginal and 21 oral Lactobacillus isolates. The concordance of both methods was at 96% with five samples discordantly identified. Fifteen different Lactobacillus species were found in the vaginal samples, primarily L. iners, L. crispatus, L. jensenii and L. gasseri. In the oral samples 11 different species were identified, mostly L. salivarius, L. gasseri, L. rhamnosus and L. paracasei. Overall, the species found belonged to six different phylogenetic groups. For several samples, MALDI-TOF MS analysis only yielded scores indicating genus-level identification. However, in most cases the species found agreed with the 16S rDNA analysis result.
Journal of Endodontics, 2014
Introduction: To date, a variety of microbial species have been isolated from endodontic infectio... more Introduction: To date, a variety of microbial species have been isolated from endodontic infections. However, endodontic clinical bacterial isolates have not been sufficiently characterized with regard to their capacity for antibiotic resistance and biofilm formation. In this study, antibiotic resistance and biofilm formation of 47 different aerobic and anaerobic bacterial isolates, belonging to 32 different species previously isolated from infected filled root canals, were studied. Methods: Antibiotic sensitivity to 11 antibiotics including penicillin G, amoxicillin, clindamycin, gentamicin, vancomycin, tetracycline, doxycycline, fosfomycin, rifampicin, ciprofloxacin, and moxifloxacin was tested using the standardized Etest method (Bio Merieux, Marcy-1'Etoile, France). The antibiotic sensitivity of 4 control strains was also estimated in parallel. Additionally, the capacity to form biofilms was quantified using the microtiter plate test. Results: Different aerobic and anaerobic bacterial species were either resistant against a number of antibiotics or showed high minimal inhibitory concentrations against clinically relevant antibiotics. Five aerobic and 2 anaerobic isolates, including Enterococcus faecalis, Streptococcus mutans, Lactobacillus fermentum, Actinomyces naeslundii, Actinomyces viscosus, Prevotella buccae, and Propionibacterium acidifaciens, were characterized as being high biofilm producers, whereas 8 aerobic and 3 anaerobic isolates were found to be moderate biofilm producers. Most isolates with resistance or markedly high minimal inhibitory concentration values were also either moderate biofilm producers or high biofilm producers. Conclusions: These results suggest that the clinical significance of endodontic infections could include that they serve as a reservoir for antibiotic resistance. Furthermore, endodontic treatment should consider the adhesion and biofilm formation by a variety of bacteria. (J Endod 2014;40:223-230)
BioMed Research International, 2014
Nature is an unexplored reservoir of novel phytopharmaceuticals. Since biofilm-related oral disea... more Nature is an unexplored reservoir of novel phytopharmaceuticals. Since biofilm-related oral diseases often correlate with antibiotic resistance, plant-derived antimicrobial agents could enhance existing treatment options. Therefore, the rationale of the present report was to examine the antimicrobial impact of Mediterranean natural extracts on oral microorganisms. Five different extracts from Olea europaea, mastic gum, and Inula viscosa were tested against ten bacteria and one Candida albicans strain. The extraction protocols were conducted according to established experimental procedures. Two antimicrobial assays-the minimum inhibitory concentration (MIC) assay and the minimum bactericidal concentration (MBC) assay-were applied. The screened extracts were found to be active against each of the tested microorganisms. O. europaea presented MIC and MBC ranges of 0.07-10.00 mg mL −1 and 0.60-10.00 mg mL −1 , respectively. The mean MBC values for mastic gum and I. viscosa were 0.07-10.00 mg mL −1 and 0.15-10.00 mg mL −1 , respectively. Extracts were less effective against C. albicans and exerted bactericidal effects at a concentration range of 0.07-5.00 mg mL −1 on strict anaerobic bacteria (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Parvimonas micra). Ethyl acetate I. viscosa extract and total mastic extract showed considerable antimicrobial activity against oral microorganisms and could therefore be considered as alternative natural anti-infectious agents.
Methods in Molecular Biology, 2014
PLoS ONE, 2012
Persistence of microorganisms or reinfections are the main reasons for failure of root canal ther... more Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa), followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and -independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis.
PLoS ONE, 2013
Residual microorganisms and/or re-infections are a major cause for root canal therapy failure. Un... more Residual microorganisms and/or re-infections are a major cause for root canal therapy failure. Understanding of the bacterial content could improve treatment protocols. Fifty samples from 25 symptomatic and 25 asymptomatic previously root-filled teeth were collected from Sudanese patients with periradicular lesions. Amplified 16S rRNA gene (V1-V2) variable regions were subjected to pyrosequencing (FLX 454) to determine the bacterial profile. Obtained quality-controlled sequences from forty samples were classified into 741 operational taxonomic units (OTUs) at 3% dissimilarity, 525 at 5% dissimilarity and 297 at 10% dissimilarity, approximately corresponding to species-, genus-and class levels. The most abundant phyla were: Firmicutes (29.9%), Proteobacteria (26.1%), Actinobacteria (22.72%), Bacteroidetes (13.31%) and Fusobacteria (4.55%). Symptomatic patients had more Firmicutes and Fusobacteria than asymptomatic patients, while asymptomatic patients showed more Proteobacteria and Actinobacteria. Interaction of disease status and age was observed by two-way ANOSIM. Canonical correspondence analysis for age, tooth restoration and disease status showed a correlation of disease status with the composition and prevalence of different members of the microbial community. The pyrosequencing analysis revealed a distinctly higher diversity of the microbiota compared to earlier reports. The comparison of symptomatic and asymptomatic patients showed a clear association of the composition of the bacterial community with the presence and absence of symptoms in conjunction with the patients' age.
Materials, 2013
Bacterial adhesion to implant biomaterials constitutes a virulence factor leading to biofilm form... more Bacterial adhesion to implant biomaterials constitutes a virulence factor leading to biofilm formation, infection and treatment failure. The aim of this study was to examine the initial bacterial adhesion on different implant materials in vitro. Four implant biomaterials were incubated with Enterococcus faecalis, Staphylococcus aureus and Candida albicans for 2 h: 3 mol % yttria-stabilized tetragonal zirconia polycrystal surface (B1a), B1a with zirconium oxide (ZrO 2 ) coating (B2a), B1a with zirconia-based composite coating (B1b) and B1a with zirconia-based composite and ZrO 2 coatings (B2b). Bovine enamel slabs (BES) served as control. The adherent microorganisms were quantified and visualized using scanning electron microscopy (SEM); DAPI and live/dead staining. The lowest bacterial count of E. faecalis was detected on BES and the highest on B1a. The
Uploads
Papers by Lamprini Karygianni