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Objective: Helicobacter pylori is known for colonizing the gastric mucosa and instigating severe upper gastrointestinal diseases such as gastritis, gastroduodenal ulcers, and gastric cancer. To date, there is no data available on the oral cavity as transmission site, whether H. pylori can survive in the oral cavity or in human saliva. The aim of the study was to investigate the influence of oral microorganisms and human saliva on the survival of H. pylori in human saliva., Methods: H. pylori strains KE, a motile derivate of type strain H. pylori 26695, and H. pylori SS1, a clinical isolate from a gastric biopsy, were grown in human pooled saliva (pooled from 4 healthy human donors, 0.22 μm filter-sterilized) or in BBF (Brucella browth formula; control) either as mono-cultures or in co-culture with Streptococcus mutans, Streptococcus oralis, Actinomyces naeslundii, Lacticaseibacillus casei and Candida dubliniensis. Bacterial survival of H. pylori and the oral microorganisms were investigated using colony forming units (CFU) assay and MALDI-TOF MS at baseline and after 24, 48 and 168 h., Results: In saliva, H. pylori KE demonstrated enhanced survival in co-culture with S. mutans, A. naeslundii, and C. dubliniensis, enduring for at least 48 h. In contrast, L. casei and S. oralis inhibited H. pylori KE in saliva. H. pylori KE could not be cultured after 168 h in saliva, neither in mono- nor co-culture. In contrast, H. pylori SS1 in saliva could be cultured after 168 h in co-culture with S. mutans and C. dubliniensis, but not in mono-culture. In BBF, H. pylori KE could be cultured after 168 h with S. mutans, L. casei and C. dubliniensis, and H. pylori SS1 with L. casei and C. dubliniensis, but not with S. mutans. Notably, the co-cultured microorganisms survived at high CFU numbers similar to those of the monocultures., Conclusion: The study suggests that H. pylori can transiently survive in human saliva and even with presence of certain oral microorganisms. However, it may not be a permanent resident of the oral microbiota. The co-survival with oral microorganisms emphasizes the necessity for studying the role of the oral microbiota in the infectious and transmission cycle of H. pylori., Competing Interests: Declarations. Ethical approval: The use of human saliva was approved by the Ethics Committee (Albert Ludwigs University of Freiburg ethics committee No. 23-1537-S1-AV). Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objectives: Low-frequency, low-intensity ultrasound is commonly utilized in various dental research fields to remove biofilms from surfaces, but no clear recommendation exists in dental studies so far. Therefore, this study aims to optimize the sonication procedure for the dental field to efficiently detach bacteria while preserving viability., Materials and Methods: Initial biofilm was formed in vivo on bovine enamel slabs (n = 6) which were worn by four healthy participants for 4 h and 24 h. The enamel slabs covered with biofilm were then ultrasonicated ex vivo for various time periods (0, 1, 2, 4, 6 min). Colony-forming units were determined for quantification, and bacteria were identified using MALDI-TOF. Scanning electron microscopic images were taken to also examine the efficiency of ultrasonications for different time periods., Results: Ultrasonication for 1 min resulted in the highest bacterial counts, with at least 4.5-fold number compared to the non-sonicated control (p < 0.05). Most bacteria were detached within the first 2 min of sonication, but there were still bacteria detached afterwards, although significantly fewer (p < 0.0001). The highest bacterial diversity was observed after 1 and 2 min of sonication (p < 0.03). Longer sonication periods negatively affected bacterial counts of anaerobes, Gram-negative bacteria, and bacilli. Scanning electron microscopic images demonstrated the ability of ultrasound to desorb microorganisms, as well as revealing cell damage and remaining bacteria., Conclusions: With the use of low-frequency, low-intensity ultrasound, significantly higher bacterial counts and diversity can be reached. A shorter sonication time of 1 min shows the best results overall., Clinical Relevance: This standardization is recommended to study initial oral biofilms aged up to 24 h to maximize the outcome of experiments and lead to better comparability of studies., (© 2023. The Author(s).)

Objective: In the last few decades, there has been a growing worldwide interest in the use of plant extracts for the prevention of oral diseases. The main focus of this interest lies in the identification and isolation of substances that limit the formation of microbial biofilm which plays a major role in the development of caries, periodontitis, and peri-implantitis. In this clinical ex vivo study, we investigated the antimicrobial effects of Rosmarinus officinalis extract against oral microorganisms within in situ initial oral biofilms., Materials and Methods: Initial in situ biofilm samples (2 h) from six healthy volunteers were treated ex vivo with R. officinalis extract at concentrations of 20 mg/ml and 30 mg/ml. The number of viable bacterial cells was determined by counting the colony-forming units. All surviving bacteria were isolated in pure cultures and identified using MALDI-TOF and biochemical testing procedures. Additionally, live/dead staining in combination with epifluorescence microscopy was used for visualizing the antimicrobial effects in the initial biofilms., Results: The number of colony-forming units in the R. officinalis-treated biofilms was significantly lower than in the untreated controls (p < 0.001). The reduction range of log10 was 1.64-2.78 and 2.41-3.23 for aerobic and anaerobic bacteria, respectively. Regarding the bacterial composition, large intra- and interindividual variability were observed. Except for Campylobacter spp., the average amount of all bacterial taxa was lower after treatment with R. officinalis than in the untreated biofilms. A total of 49 different species were detected in the untreated biofilms, while only 11 bacterial species were detected in the R. officinalis-treated biofilms. Live/dead staining confirmed that the R. officinalis-treated biofilms had significantly lower numbers of surviving bacteria than the untreated biofilms., Conclusions: The treatment with R. officinalis extract has a significant potential to eliminate microbial oral initial biofilms., Clinical Relevance: The results of this study encourage the use of R. officinalis extracts in biofilm control and thus in the treatment of caries and periodontitis as a herbal adjuvant to synthetic substances., (© 2022. The Author(s).)

Objective: The persistence of pathogenic microorganisms in root canals is the most common reason for the failure of root canal treatment and the necessity of a root filling treatment, which results in an uncertain prognosis due to technical complexity and the variety of highly adaptable microorganisms. This study evaluated the effect of passive ultrasonic irrigation (PUI) on the outcome of the microbial analysis of root canal-treated teeth with persistent or recurrent apical inflammation in vivo., Materials and Methods: Sample collection was performed after root filling removal (sample S1, control group) and after PUI with NaCl (sample S2) using sterile paper points. In total, 19 samples were obtained. Quantification was performed by means of serial dilution of the samples. Subcultivated pure cultures were identified using MALDI-TOF MS complemented by the Vitek-2-System or PCR, followed by sequencing of the 16S rRNA gene. The results of the samples (S1 and S2) were evaluated regarding their bacterial count and composition., Results: The total count of bacteria and the number of aerobic/facultative anaerobic microorganisms significantly increased in the S2-samples after application of PUI. The number of obligate anaerobic microorganisms showed an increase after PUI, although it was not significant. We detected 12 different aerobic/facultative anaerobic microorganisms before PUI, and in 21 cases after PUI. Two different obligate anaerobic microorganisms were found in S1 samples compared to nine different species in S2 samples., Conclusions: PUI is a powerful method for detaching bacteria in infected root canals and enables a more precise analysis of the etiology of persistent endodontic infections., Clinical Relevance: This study indicates that PUI exerts a positive cleansing effect and adds to the accessibility of microorganisms during the application of bactericidal rinsing solution in root canal treatments., (© 2022. The Author(s).)

Objectives: The aim of the present study was to evaluate the growth and acid synthesis of cariogenic pathogens in vitro when rebaudioside A was the sole source of carbon, compared to sucrose, xylitol, and a commercial sweetener containing rebaudioside A., Materials and Methods: First, sucrose or sugar substitutes were added to human saliva and the pH was measured over 10 h. Subsequently, the growth and acid synthesis efficiency of Streptococcus mutans, Streptococcus sobrinus, Streptococcus oralis, Lactobacillus rhamnosus, Lactobacillus paracasei, and Candida albicans were investigated: after adding them to minimal culture media containing only one of the carbon sources, the optical density and pH were determined over 10 h., Results: A significant pH drop only occurred in the sucrose group of human saliva whereas no differences were found between the sugar substitute groups. The individual strains of caries pathogens showed significantly lower growth rates and less acid synthesis in the sugar replacement groups, in comparison to sucrose. However, these groups also showed bacterial growth and a decline in pH., Conclusion: Xylitol appeared to be superior to rebaudioside A regarding the inhibition of mutans streptococci. The results for the remaining microorganisms indicate that rebaudioside A causes a similar or lower acid synthesis than xylitol. As saliva contains buffer systems, the metabolism of rebaudioside A did not seem to shift the equilibrium to demineralization., Clinical Relevance: Rebaudioside A may not be suitable as a targeted Streptococcus mutans treatment, but it could contribute to a reduction of caries risk as a replacement for sucrose.

Background: Mixed cell populations from oral tissues may be superior to pure stem cells for regenerative approaches. Therefore, the aim of the present study was to explore the osteogenic potential of mixed cells derived from oral connective tissues compared to alveolar osteoblasts., Materials and Methods: Primary cells were isolated from the alveolar bone, periodontal ligament and gingiva. Following characterization by colony formation, growth capacity and flow cytometry, all cells were subjected to osteogenic differentiation induction and screened for a large panel of osteogenic markers using western blots, qPCR arrays, and matrix mineralization and alkaline phosphatase quantification., Results: Non-induced mixed cells from gingiva showed higher colony formation efficiency but decreased proliferation compared to non-induced periodontal mixed cells, while both entities revealed similar surface markers tested in this setup. Following osteogenic induction, all cell populations individually expressed receptors with distinctively activated downstream effectors. Gene expression of induced periodontal mixed cells was similar to alveolar osteoblasts, but was differently modulated in gingival mixed cells. The latter failed to achieve osteogenic differentiation in terms of matrix mineralization and alkaline phosphatase activity, which was well observed in periodontal mixed cells and osteoblasts., Conclusion: Mixed cells from periodontal ligament but not from gingiva feature an inherent osteogenic capacity in vitro. From these results, it can be concluded that periodontal cells do not require further stem cell enrichment in order to qualify for bone regeneration., Clinical Relevance: Our data contribute to the development of novel cell-based therapies using mixed cells from the periodontal ligament in regenerative periodontics.

Objective: We examined whether fluoride/vitamin D supplementation in the first year of life is associated with caries or molar incisor hypomineralization (MIH) at 10 years of age., Methods: The study population consisted of 406 children for whom information on fluoride/vitamin D supplementation during the first year of life was available. Dental examination at the age of 10 included caries and MIH registration. The results of logistic regression models were adjusted for gender, age, BMI, parental education, and equivalent income., Results: Children receiving supplementation during the entire first year of life had a significantly lower probability of having caries-related restorations in primary teeth in comparison to those who received supplementation for less than 6 months (fluoride supplementation: odds ratio (OR) for d 3-4 mfs 2.47 (1.32-4.63), for fs 2.70 (1.43-5.10); vitamin D supplementation: OR for d 3-4 mfs 2.08 (1.00-4.32), fs 2.50 (1.19-5.25)). The majority of logistic regression analyses indicated no association between supplementation and MIH., Conclusions: It was found a consistent significant caries-preventive effect in the primary dentition of children who received fluoride (256/372)/vitamin D supplementation (274/376) in all 12 months over the first year of life; no effects were observed for permanent dentition. The high parental interest in supplementation is linked to an imbalance of the study groups. Furthermore, tooth brushing frequency, use of fluoride toothpastes and/or other oral hygiene products were not recorded during the observation period which may also confound the results., Clinical Relevance: Fluoride/vitamin D supplementation can be used in children for preventing caries in the primary dentition.

Objectives: The correlation between caries and the oral prevalence of Candida spp. in children is contradictory in literature. Thereby, authors focused on Candida albicans as the most isolated Candida species from the oral cavity. Therefore, the aim of the present study was to compare caries-free and caries-bearing children regarding their oral carriage of Candida spp., Material and Methods: Twenty-six caries-free (CF group) and 26 caries-active children (CA group) were included into this study. Three different types of specimens were assessed, saliva and plaque, and in the case of caries, infected dentine samples were microbiologically analyzed for aerobic and anaerobic microorganisms and their counts. Special attention was given to the differentiation between C. albicans and Candida dubliniensis. Additionally, different biochemical tests, VITEK 2 (VITEK®2, bioMérieux, Marcy-l'Etoile, France) and 16S and 18S ribosomal DNA (rDNA) sequencing, were applied for identification., Results: The detection of C. albicans did not differ between the CF and CA groups. C. dubliniensis was never detected in any specimen of the CF group, but occurred in one quarter of the CA group (27 % in plaque, 23 % in saliva), thus leading to a statistically significant difference between the two groups (p < 0.05). In six of these cases, C. dubliniensis was detected concomitantly in saliva and plaque and once only in plaque. CA group harbored statistically more Streptococcus mutans than the control group revealing a correlation between S. mutans and C. dubliniensis regarding the caries group., Conclusions: This is the first study reporting a frequent detection of C. dubliniensis in caries-active children, which could have been underestimated so far due to difficulties in differentiation between this yeast species and C. albicans., Clinical Relevance: Microbiological diagnostic-especially of oral Candida species-is an important determinant for identifying etiological factors of dental caries in children.

Objectives: 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., Materials and Methods: 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)., Results: Overall, A and B presented the lowest CFU values for all microorganisms, while C sheltered significantly less E. faecalis, P. aeruginosa, and C. albicans than BES. Compared to the controls, B demonstrated the lowest vitality values for E. coli (54.12 %) and C. albicans (67.99 %). Interestingly, A (29.24 %) exhibited higher eradication rates for S. aureus than B (13.95 %)., Conclusions: Within the limitations of this study, a-C:H:N-coated 3Y-TZP surfaces tended to harbor less initially adherent microorganisms and selectively interfered with their vitality., Clinical Relevance: This could enable further investigation of the new multi-functional zirconia surfaces to confirm their favorable antimicrobial properties in vivo.

Objectives: The aim of this study was to evaluate the antimicrobial effect of gaseous ozone compared to conventional methods against Enterococcus faecalis., Materials and Methods: One hundred twenty-five teeth were infected by E. faecalis and were incubated for 72 h to form biofilm. Teeth were distributed among five groups. In the first group, ozone was used; in the second group, teeth were rinsed with 20 % ethylenediaminetetraacetic acid (EDTA); in the third group, with 3 % sodium hypochlorite (NaOCl). Group 4 combined 20 % EDTA with ozone. NaOCl and ozone were combined in group 5. After treatment, the samples with paper points were taken, followed by dentin samples taken with K-file, and cultured for 24 h. Then bacterial colonies were counted., Results: All treatments reduced significantly (p < 0.05) the bacteria. Paper points' samples showed 85.38 % reduction after ozone. The highest reduction was observed in NaOCl group (99.98 %). EDTA reduced bacteria by 80.64 %. Combination of NaOCl and ozone eradicated 99.95 % of the bacteria. Combination of EDTA and ozone reduced E. faecalis up to 91.33 %. The dentin chips showed the following: the highest CFU counts were observed in EDTA group, followed by ozone and NaOCl group. The lowest CFU counts were found in NaOCl-ozone group and EDTA-ozone group., Conclusions: Ozone reduced E. faecalis, even organised in a biofilm, however, lower than NaOCl. No treatment reduced totally the bacteria., Clinical Relevance: Used as an adjuvant, ozone can increase the efficacy of conventional rinsing like EDTA and presents an alternative treatment when NaOCl cannot be used e.g. in teeth with a wide-open apical foramen.

Objectives: To investigate the effect of a zinc oxide/zinc sulphate-based cement as a temporary filling material during root canal treatment on the occurrence of cracks within the filling material or the tooth., Materials and Methods: Root canals of 122 extracted human molars were prepared using ProTaper instruments. Standardized occlusal-distal cavities were prepared. After placing calcium hydroxide into the root canals, the teeth were divided randomly into four groups containing 33 specimens each. In the Coltosol group, the cavity was completely filled with Coltosol® F. In the Coltosol-Clearfil group, a 2-mm layer of Coltosol® F was placed into the apical part of the pulp chamber, and in the Clearfil group, a foam pellet was placed into the coronal pulp chamber. The remaining cavities were filled with Clearfil™. In the control group, the cavities were not restored. The teeth were stored at 37 °C for 14 days and examined every 24 h under a stereomicroscope., Results: Fractures of the filling and/or the tooth were only observed in the Coltosol group. All Coltosol® F restorations had cracks after 24 h. Tooth fractures were found in 25 (76 %) teeth. Among these teeth, 21 (84 %) had crown fractures, four (16 %) had root-crown fractures. All root-crown fractures were vertical., Conclusion: Coltosol® F when used alone led to tooth fractures in two-surface class II cavities in teeth undergoing root canal treatment., Clinical Relevance: Coltosol® F solely used as restorative material may lead to tooth fractures in two-surface class II cavities.

Objectives: Root canal treatment failures often correlate with persistent biomaterial-associated endodontic infections. The aim of the present study was to assess the impact of endodontic obturation material sampling from root canals with posttreatment apical periodontitis on improving standard study protocols., Materials and Methods: Samples from previously filled root canals and their corresponding endodontic filling materials were obtained from five root-filled teeth with posttreatment periradicular lesions. After cultivation, the isolated microorganisms were quantified and biochemically identified. Moreover, clone libraries were constructed after the amplification of bacterial 16S ribosomal DNA (rDNA) from the same samples. DNA from selected clones was sequenced to identify microbial species. Transmission electron microscopy (TEM) aided visualization of the detected bacteria., Results: Overall, 22 taxa of the phyla Firmicutes, Actinobacteria, and Bacteroidetes were detected in both obturation and root canal samples by culture-dependent and culture-independent methods. Root canal fillings sheltered 17 species (3.30-7.50 × 10(3) CFU/ml). Of these, nine were detected solely in the retrieved obturation materials. The reinfected root canals harbored 13 taxa (3.48-7.36 × 10(3) CFU/ml). Obligate and facultative anaerobic bacteria prevailed. The number of different species ranged from 1 to 5 within a single sample. Fungi were not detected., Conclusions: Bacteria can colonize both root canals and endodontic fillings in vivo., Clinical Relevance: Integrating the sampling of obturation materials with standard root canal sample collection offers a clearer insight into the actual microbial flora of reinfected root canals and improves the study protocols of secondary/persistent endodontic infections.

Objectives: Interdental cleaning is an essential component of home plaque control to prevent periodontitis and caries. There is limited data on the efficacy of commonly used metal-core interdental brushes in comparison to metal-free interdental brushes. The aim of this study was to compare a new rubber interdental bristle (Fuchs®) with a standard metal-core interdental brush (TePe®) for their impact upon gingival bleeding, plaque removal, and patient experience., Materials and Methods: A single-blind, prospective, randomized, and controlled clinical trial with a crossover design was used to measure plaque index (Turesky-Modified Quigley & Hein Index), bleeding index (Eastman Interdental Bleeding Index by Caton & Polson), and patient satisfaction by means of questionnaires in 39 patients. Each patient was randomly assigned with regard to the sequence of interdental product used and recalled., Results: Both groups showed statistically significant decreases of plaque after a single usage, respectively. Bleeding was statistically significantly reduced after 4 weeks, with no statistically significant differences concerning between the two tested interdental brushes. Rubber interdental bristles reached significantly higher scores with regard to patient acceptance in overall assessment and in sub-items for less pain during usage, comfort of brushing, and willingness to buy the product., Conclusion: Rubber interdental bristles were similarly effective compared to the interdental brushes. In addition, rubber interdental bristles were significantly more comfortable for participants than metal-core brushes., Clinical Relevance: Rubber interdental bristles can be used as an alternative interdental cleaning product which may be more accepted by patients.

Objectives: The aim of this study was to evaluate the accuracy of two-dimensional (2D) and three-dimensional (3D) root canal length measurements in molar teeth using cone beam computed tomography (CBCT)., Materials and Methods: Root canal lengths of twenty molar teeth were measured with two different CBCT approaches. After adjusting the CBCT images, 2D measurements were performed within the sagittal plane between the apical foramen and the coronal reference (cusp). The 3D approach measured centrically in axial planes. A linear mixed model with random intercepts was fitted to compare differences between methods (2D and 3D). The correlation between CBCT measurements and the actual root canal length was evaluated using the Pearson correlation coefficient., Results: Differences between 3D measurements and the actual root canal lengths were significantly smaller compared to the 2D approach (p < 0.001). Mean differences were 0.32 and 0.58 mm, respectively. A high correlation was found between the actual root canal length and 3D measurements (Pearson correlation coefficient = 0.97). Compared to the actual root canal length, 80 % of the 3D measurements were within the limits of ±0.5 mm., Conclusions: 3D measurements of root canals in molar teeth are more accurate than simple 2D measurements and show a high correlation to the actual lengths., Clinical Relevance: In cases where a CBCT is already available, root canal lengths in molar teeth can be accurately predetermined using a standardized 3D approach.

Objectives: Caries excavation is still a demanding procedure today. In the past years, methods using light-induced fluorescence have proven their ability to detect bacteria remaining in the tooth's hard tissues. However, methods to control the complete removal of infected dentine have not yet been validated. The aim of the present study was to evaluate the degree of dentine removal during caries excavation using a camera- and software-based device as a guide for fluorescence-aided caries excavation (FACE) in comparison to visual-tactile inspection and a dye-staining method., Materials and Methods: One hundred teeth with carious lesions (C3) were randomly allocated to four groups and excavated with the respective method as primary excavation control method. When the first method indicated complete caries excavation, a second method was used to affirm this. Subsequently, dentine samples were taken to determine residual contamination using PCR. After embedding and sectioning the teeth dentine, microhardness was measured perpendicular to the cavity floor., Results: By using the camera- and software-based device, 94 % of the cavities tested were free of bacterial contamination. All cavities excavated with the dye-staining or visual-tactile control method were free of bacterial contamination but resulted in a significantly higher dentine removal., Conclusion: The camera- and software-assisted caries excavation seems to be an adequate method to remove contaminated dentine without removing affected dentine., Clinical Relevance: Beside its original application to monitor carious, the device is safe, effective, and easy to use for guided caries excavation and is of worth for both everyday practice and undergraduate education.

In the context of cell-based oral hard tissue regeneration, especially assumed plasticity of oral host tissue cells in response to human mesenchymal stem cells (hMSCs), is poorly understood. To investigate this area, we assess osteogenic features in various oral cell types during hMSC coculture, including human alveolar osteoblasts (hOAs), periodontal ligament cells (hPDLs) and gingival fibroblasts (hGFs). Interactive hMSC coculture globally enhanced the transcription of osteogenic genes, in all oral cell types under study, as revealed by qRT-PCR and did not affect oral cell proliferation compared with controls in a transwell coculture system as evaluated by 5-bromo-2'-deoxyuridine proliferation assay. 3D gel-derived hMSC cocultures exhibited an abundance of bone-related key molecules in oral cells, which followed the ranking hOAs > hGFs > hPDLs. Compared to matched controls, this hierarchy also applied for the presence of higher amounts of extracellular matrix deposits and mineralization nodules in interactive hMSC coculture. Our results show for the first time that in the context of prospective periodontal tissue regeneration strategies, hMSCs influence oral cells by gradually shaping their plasticity, particularly features associated with an osteogenic phenotype. These novel findings contribute another piece to the conceptual hMSC action puzzle and valuably support the notion that hMSCs trigger osteogenesis in the oral cell context.

Enterococcus faecalis (E. faecalis) is a Gram-positive bacterium, mostly recovered from root-filled teeth with persistent periapical lesions. Bacterial contamination of root canals inevitably results in interaction between E. faecalis and periapical tissues during the dynamic process of periapical inflammation. This study investigated the impact of heat-inactivated endodontic E. faecalis on the proliferation and the differentiation of ovine osteoblast-like cells, in an attempt to elucidate its putative enhanced pathogenicity mechanisms. Therefore, two different concentrations of a heat-inactivated endodontic E. faecalis isolate (2 × 10(6) or 2 × 10(8) CFU/ml) were incubated with ovine osteoblast-like cells for 7 and 14 days, respectively. Cells without antigen served as control. The effects of antigen on cell growth were evaluated by a proliferation assay (EZ4U). Furthermore, the assessment of alkaline phosphatase (ALP) activity, calcium deposition, and osteocalcin (OCN) gene expression through quantitative real-time PCR determined the degree of osteogenic cell differentiation. Scanning electron microscopy (SEM) was also performed to detect alterations in cell morphology. Interestingly, although highly concentrated E. faecalis increased cellular reproduction after 14 days, ALP activity and OCN gene expression decreased in an antigen concentration-dependent and incubation time-independent way. SEM images revealed E. faecalis adhesion on cells, a fact that might contribute to its virulence. These results suggest that E. faecalis stimulated cell multiplication, whereas it likely restrained cell differentiation of ovine osteoblast-like cells. In conclusion, the presence of E. faecalis in root canals may negatively affect periapical new bone formation, and thus, the healing of periapical lesions.

Various situations encountered by a clinician during the daily routine including surgical periodontitis therapy, dental implant insertion, or tooth extraction involve the contact of saliva with the jaw bone. However, there are only sparse data concerning the influence of saliva on bone cells. Saliva specimens were incorporated within culture medium and administered to murine MC3T3 osteoblasts, of which the morphology (REM), proliferation (EZ4U), and differentiation (qRT-PCR, alkaline phosphatase activity, extracellular matrix calcification) were assessed. Simultaneously, the composition of saliva media was analyzed with respect to the content of lactoferrin, activities of classical salivary enzymes, and the ability to provoke inflammatory cytokine production (enzyme-linked immunosorbent assay) in MC3T3 osteoblasts. The morphology, proliferation, and expression of differentiation-associated genes were seriously handicapped by saliva contact. Saliva-touched cells exhibited less alkaline phosphatase but normal levels of extracellular matrix mineralization. Saliva-containing culture media featured physiological activities of salivary enzymes and considerable amounts of lactoferrin but almost completely lacked salivary alkaline phosphatase and unspecific proteases. Upon saliva incubation, MC3T3 osteoblasts did not release noteworthy levels of interleukin-1 beta or tumor necrosis factor alpha. Although saliva is generally considered to vitalize oral tissues, this study reveals that it harms osteoblast-like cells more due to the presence of salivary enzymes than by triggering of inflammation. This issue is clinically relevant because it broadens the understanding of the bone cell fate within the rather complex cosmos of the oral cavity thereby providing a basis for clinical decision making and treatment guidelines. It seems to be reasonable to restrict the contact period between saliva and bone.

It was the aim of the study to evaluate the clinical and antibacterial effect of a dentifrice containing an anti-inflammatory plant extract (SB) versus a placebo (PLA) using an experimental gingivitis model. Forty subjects (20 per group) discontinued all oral hygiene measures for four teeth for a period of 21 days using a shield (to generate a possible gingivitis) while they could brush the other teeth normally. After brushing, the shield was removed and teeth were treated with the randomly assigned toothpaste slurry for 1 min. Löe and Silness gingival index (GI), Silness and Löe plaque index (PI), and biofilm vitality (VF%) were assessed at days 0, 14, and 21, respectively. Subjects of the PLA group developed a GI of 0.82 ± 0.342 (day 14) and 1.585 ± 0.218 (day 21), while the data of the SB group were significantly reduced (0.355 ± 0.243 and 0.934 ± 0.342, p < 0.001). While PI was significantly reduced at all follow-up appointments, reductions in VF reached the level of significance only at day 21. The results suggest that the new toothpaste formulation was able to significantly reduce the extent of gingivitis, plaque development, and vital flora.

The aim of the study was to monitor changes in oral health and oral biofilm composition in vivo during an experiment simulating prehistoric lifestyle and diet and poor oral hygiene. Thirteen subjects lived for a period of 8 weeks under Neolithic conditions. The following clinical parameters were recorded before and after the project: gingival and plaque index (Löe and Silness, Acta Odontol Scand 21:533, 1963; Silness and Löe, Acta Odontol Scand 22:121-135, 1964), probing pocket depth, and bleeding upon probing. In addition, supragingival plaque samples were collected both before and after the project and were analysed quantitatively using multiplex fluorescence in situ hybridization and confocal laser scanning microscopy. The following plaque bacteria were evaluated: Streptococcus spp., Veillonella spp., Fusobacterium nucleatum, and Actinomyces naeslundii. The plaque index increased significantly from 1.12 up to 1.55 over the 8-week period (gingival index before, 0.46; after, 0.93; p < 0.05). A strong correlation of both indices was recorded before (r = 0.77) and after (r = 0.83) participation in the study. Each of the children in the study showed a progression of carious lesions and/or new areas of demineralisation. The probing pocket depth and bleeding upon probing were not affected. All subjects yielded an intra-individual shift in biofilm composition. The proportion of F. nucleatum decreased across all subjects. The proportion of Veillonella spp. increased among the children. Poor oral hygiene and change of diet lead to an increase in oral plaque and gingival inflammation. The inter-individual comparison indicated a shift in bacterial composition.

The aim of the present study was to evaluate the remineralization potential of five dentifrices with different fluoride concentrations. Initial caries lesions were created in 72 cylindrical enamel blocks from deciduous teeth. The specimens were randomly distributed among six experimental groups corresponding to six experimental periods. Each of the six volunteers carried two deciduous enamel specimens fixed in an intraoral appliance for a period of 4 weeks. They brushed their teeth and the enamel blocks at least two times a day with dentifrices containing 0 ppm (period 1), 250 ppm (period 2), and 500 ppm fluoride (period 3), respectively. A second group of volunteers (n = 6) used dentifrices with a fluoride content of 0 ppm (period 4), 1,000 ppm (period 5), or 1,500 ppm (period 6). At the end of the respective period, the mineral content was determined by transversal microradiography (TMR). The use of dentifrices containing 500 ppm fluoride (38% MR), 1,000 ppm fluoride (42% MR), and 1,500 ppm fluoride (42% MR) resulted in a statistically significant higher mineral recovery compared to the control group (0 ppm fluoride). Mineral recovery was similar after use of dentifrices containing 0 and 250 ppm fluoride (24%; 25%). It is concluded that it is possible to remineralize initial carious lesions in deciduous enamel in a similar way as it has been described for enamel of permanent teeth.

The aim of this double-blind, controlled crossover study was to evaluate the influence of food preservatives on in situ dental biofilm growth. Twenty-four volunteers wore appliances with six specimens each of bovine enamel to build up intra-oral biofilms. During three test cycles, the subjects had to put one half of the appliance twice a day in one of the assigned active solutions (0.1% benzoate, BA; 0.1% sorbate, SA or 0.2% chlorhexidine, CHX) and the other into NaCl. After 5 days, the developed biofilms were stained with two fluorescent dyes to visualise vital (green) and dead bacteria (red). Biofilms were scanned by confocal laser scanning microscopy and biofilm thickness (BT) and bacterial vitality (BV%) were calculated. After a washout period of 7 days, a new test cycle was started. The use of SA, BA and CHX resulted in a significantly reduced BT and BV compared to NaCl (p<0.001). Differences between SA and BA were not significant (p>0.05) for both parameters, while CHX showed significantly lower values. Both preservatives showed antibacterial and plaque-inhibiting properties, but not to the extent of CHX. The biofilm model enabled the examination of undisturbed oral biofilm formation influenced by antibacterial components under clinical conditions.

This in vitro study evaluated the influence of one halogen and two light-emitting diode (LED) curing units on the curing depth of a conventional hybrid and two translucent resin composites by measuring the Knoop microhardness. In the first part of the study, a conventional hybrid resin composite and three curing units (one halogen: 40 s polymerization time, two LEDs: 10 and 20 s) were used. Ten cylindrical resin composite samples were prepared for each curing unit and each polymerization time tested. After polymerization, the soft part of the samples was removed. The samples were embedded in a polyacrylic resin and separated in the middle towards the direction, top-bottom. On the section plane, Knoop microhardness measurements were performed every 1 mm, starting at 0.5 mm under the surface. In the second part of the study, two translucent resin composites and a conventional hybrid composite resin were cured with the three curing units, and the microhardness was measured as mentioned above. The difference between the curing units tested was found statistically significant (p = 0.0009), as well as the difference between the materials concerning curing depth (p = 0.0001). Both translucent materials achieved microhardness values equal to the 80% of the surface values, in depths 3.5-5.5 mm, depending on the curing units used.

The objective of this study was to analyse fluoride uptake and microhardness alteration of carious-like demineralised enamel after application of differently concentrated acidulated sodium fluoride gels and to determine the effect of fluoridation on enamel resistance against subsequent demineralisation. Artificial caries-like lesions of bovine enamel specimens were treated with sodium fluoride gels of different concentration (group A: 1.25%, group B: 0.62%, group C: 0.31%, group D: 0.15%; n=20 each group) for 5 min and stored in artificial saliva for 24 h. This cycle was carried out three times. Subsequently, KOH-soluble and structurally bound fluoride (determined at depths of 30, 60, and 90 microm) were analysed. In the second part of the study, for each 12 enamel specimens surface microhardness was determined before and after demineralisation, after fluoridation with the differently concentrated gels A-D, and after a second demineralisation. With all groups uptake of KOH-soluble and structurally bound fluoride resulted in higher levels than baseline content. Statistical analysis revealed significant differences between fluoride uptake among the groups, with highest uptake for the 1.25% gel and lowest for the 0.15% gel. Moreover, with all gels highest uptake was observed in the outermost enamel layer (P<0.05). Microhardness values after second demineralisation increased with increased concentration of the applied sodium fluoride gel. Increasing concentration of the applied gel implies better protection of the enamel specimens against subsequent demineralisation (P<0.05). It is concluded that differently concentrated acidulated sodium fluoride gels resulted in concentration-related significant uptake of fluoride in carious-like demineralised enamel, leading to a better demineralisation protection with increasing fluoride concentration in the gel.

The purpose of this double-blind study was to evaluate the effectiveness of a commercially available fluoride lacquer (Bifluorid 12) containing CaF2 (6%) and NaF (6%) in reducing dentine hypersensitivity. A fluoride lacquer containing only NaF (6%) served as a control. Twenty-five adult patients complaining about at least two hypersensitive teeth participated in this study. In each patient and at each appointment, one tooth was treated with Bifluorid 12, while the other was treated with the control substance. Sensitivity levels were determined before and after the application of each lacquer at baseline as well as at 1, 2 and 3 weeks after the start of study. The final evaluation of hypersensitivity was performed at 4 weeks, and follow-ups were undertaken at 6 and 12 months. A reproducible air blast stimulus and a visual analogue scale were used for evaluation. Results demonstrated a distinct reduction of hypersensitivity after 1, 2 and 3 weeks in the Bifluorid 12 group. Initially, no obvious effects could be observed in the control group. However, a clear alleviation could be observed after 2 and 3 weeks with the control. After 4 weeks, the overall sensitivity scores were comparably low, without any significant differences between the two fluoride lacquers. In both groups, the effects of treatment were seen over the 12-month observation period. Bifluorid 12 was considered at least comparable to the control. It is concluded from this study that Bifluorid 12 is effective in the initial reduction of dentine hypersensitivity. The combination of CaF2/NaF can be recommended for clinical use.

Stereotactic endoscopic techniques are extremely helpful in diagnosis and therapy of cystic intracerebral space occupying lesions. Acute space occupying lesions can be managed effectively and without major tissue traumatization. Up to now we have operated on more than 70 cystic intracerebral space occupying lesions with a stereotactic endoscopic technique. The main diagnoses were colloid cysts, cystic craniopharyngeoma, arachnoidal and pineal cysts. In must be stressed that in cystic anaplastic astrocytomas and glioblastomas as well as metastases only an acute inner cerebral decompression can be achieved by neuroendoscopic techniques in combination with the application of reservoir systems. In benign parenchymal or intraventricular cysts neuroendoscopic intervention is performed for definitive treatment. The results are overall encouraging. There was no operative mortality and operative morbidity was below 3%. Postoperative follow-up in patients with benign cysts showed no evidence of recurrence.

Interstitial curietherapy with 125-Iodine is an effective therapeutic option in the treatment of low grade gliomas. Four cases with astrocytoma grade II are presented, where tumour growth characteristics have changed to anaplasia during interstitial irradiation after a primary period of tumour regression. Anaplastic transformation could be due to a radiation effect or an insufficient therapeutic influence of interstitial irradiation on natural tumour progression of glioma growth due to genetic events.