Axel Spahr

Characteristic distribution pattern of Helicobacter pylori in dental plaque and saliva detected with nested PCR.

The precise mode of transmission and the natural reservoir for Helicobacter pylori are unknown. PCR assays have proved to be highly sensitive and specific and are regarded as the method of choice for detecting H. pylori DNA in the oral cavity. The aim of this study was to investigate the prevalence and distribution of H. pylori in the oral cavity. Forty-two patients undergoing gastroscopy were investigated for the presence of H. pylori in dental plaque and saliva by nested PCR, and in the stomach by the 13C-urea breath test. Samples tested comprised dental plaque from molars, premolars and incisors and saliva. Two sets of primers homologous to the 860-bp fragment of H. pylori DNA, which have been shown previously to be highly sensitive and specific, were used for nested PCR. Eleven patients (26.2%) were infected with H. pylori in the stomach. H. pylori DNA was identified in dental plaque samples from 41 patients (97%) and in 23 saliva samples (55%). The prevalence in dental plaque from molars, premolars and incisors was 82%, 64% and 59%, with an odds ratio of 3.18, 1.24 and 1 (reference), respectively. In conclusion, H. pylori was present in the oral cavity of 97% of tested patients, with a characteristic distribution that was independent of the infection status of the stomach. Thus H. pylori may belong to the normal oral microflora.

Additive Antimicrobial Activity of Calcium Hydroxide and Chlorhexidine on Common Endodontic Bacterial Pathogens

It is an established procedure to use calcium hydroxide [Ca(OH)2] as a therapeutic component to achieve sterilization of infected root canals. Unfortunately, some bacterial species are relatively resistant to Ca(OH)2 and are therefore associated with treatment failures. The objective of this study was to identify combinations of substances that improve antimicrobial activity. Ca(OH)2 was used as a suspension or as a component of gutta-percha points. Efficacy of killing was compared for (a) Ca(OH)2 suspension alone; (b) zinc oxide (ZnO) points alone; (c) Ca(OH)2 suspension combined with ZnO points; (d) Ca(OH)2 suspension combined with ZnO/chlorhexidine points; and (e) Ca(OH)2 points combined with ZnO/chlorhexidine points. Common endodontopathogenic bacterial species (Enterococcus faecalis, Fusobacterium nucleatum, Peptostreptococcus micros, Porphyromonas gingivalis, Streptococcus intermedius) in pure cultures served as target organisms. In the in vitro assay, the antibacterial substances and approximately 10(7) colony forming units of test bacteria were co-incubated in diluted human serum under growth conditions appropriate for the single bacterial species. Samples were taken on days 0, 1, 2, 4, 7, and 14 of incubation and viable counts determined. Gram-negative bacteria were sufficiently killed by Ca(OH)2 alone. Combinations with other agents did not improve the killing kinetics. For the inactivation of the Gram-positive bacteria P. micros and S. intermedius, a combination of Ca(OH)2 and ZnO/chlorhexidine, killed the bacteria faster than Ca(OH)2 alone. For E. faecalis, combining Ca(OH)2 suspension with ZnO/chlorhexidine points lead to a faster decrease in the overall number of viable bacteria, but no test conditions lead to the complete loss of culture viability. The results of this study support the use of a combination of Ca(OH)2 and chlorhexidine for faster eradication of P. micros and S. intermedius from infected root canals.

Helicobacter pylori in the oral cavity: high prevalence and great DNA diversity.

To test the hypothesis that Helicobacter pylori may be transmitted by the oral–oral route, we applied nested PCR and DNA sequencing to detect and analyze H. pylori DNA in the oral cavity of 20 adult patients undergoing endoscopy. Dental plaques of molars, premolars, and incisors and saliva were collected. Additional paraffin-embedded gastric biopsies were analyzed in four patients. Two sets of highly sensitive and specific primers, EHC-U/EHC-L and ET5-U/ET-5L directed to a 860-bp fragment of H. pylori DNA, were used in the nested PCR. Eight patients had an active infection in the stomach determined with the [13C]urea breath test and the other 12 were negative. Nested PCR showed that all 20 subjects (100%) were positive for H. pylori in the oral cavity. DNA sequencing demonstrated that all tested PCR products of the expected size from the oral samples have more than 97% identity with that from H. pylori type strain ATCC 43629. However, sequences differed in oral samples from different subjects as well as between different oral locations and gastric biopsies within the same individuals. In conclusion, the oral cavity may be a permanent reservoir for H. pylori and can harbor multiple H. pylori strains at the same time.

Feasibility of ultra-short echo time (UTE) magnetic resonance imaging for identification of carious lesions.

The objective of this study was to investigate the potential of ultra short echo time imaging for the assessment of caries lesions and early demineralization. 12 patients with suspected caries lesions underwent a dental magnetic resonance imaging investigation comprising ultra short echo time imaging (echo time = 50 μs) and spin echo imaging. Before the dental magnetic resonance imaging, all patients underwent a conventional clinical dental investigation including visual assessment of the teeth as well as dental x‐ray imaging. All lesions identifiable in the x‐ray could be clearly identified in the ultra short echo time images, but only about 19% of the lesions were visible in the spin echo images. In 19% of all lesions, the lesions could be more clearly delineated in the ultra short echo time images than in the x‐ray images. This was especially the case for secondary lesions. In direct comparison with the x‐ray images, all lesions appeared substantially larger in the dental magnetic resonance imaging data. The presented data provide evidence that caries lesions can be identified in ultra short echo time magnetic resonance imaging with high sensitivity. The apparent larger volume of the lesions in dental magnetic resonance imaging may be attributed to fluid accumulation in demineralized areas without substantial breakdown of mineral structures. Magn Reson Med, 2011.

Bioinformatic analysis and molecular modelling of human ameloblastin suggest a two-domain intrinsically unstructured calcium-binding protein

Ameloblastin (AMBN) was originally believed to be an enamel-specific extracellular matrix glycoprotein secreted by ameloblasts. Recently, AMBN expression was also detected in developing mesenchymal dental hard tissues, in trauma-induced reparative dentin, and during early craniofacial bone formation. The function and structure of AMBN still remain ambiguous, and there are no known proteins with similar primary sequences. We therefore performed a bio-informatic analysis of AMBN to model ab initio the three-dimensional structure of the molecule. The results suggest that AMBN is a two-domain, intrinsically unstructured protein (IUP). The analysis did not reveal any regions with structural similarity to known receptor-ligand systems, and did not identify any higher-order structures similar to functional regions in other known sequences. The AMBN model predicts 11 defined regions exposed on the surface, internalizing the rest of the molecule including a human-specific insert. Molecular dynamics analysis identified one specific and several non-specific calcium-binding regions, mostly at the C-terminal part of the molecule. The model is supported by previous observations that AMBN is a bipolar calcium-binding molecule and hints at a possible role in protein-protein interactions. The model provides information useful for further studies on the function of AMBN.

Ameloblastin promotes bone growth by enhancing proliferation of progenitor cells and by stimulating immunoregulators

In this study, we examined the role of the enamel matrix protein, ameloblastin, in bone growth and remodelling, and attempted to identify some of the molecular mechanisms involved in these processes. The effects of recombinant ameloblastin (rAmbn) were tested in vivo in rats, and in vitro in primary human mesenchymal stem cells, osteoblasts, chondrocytes, and osteoclasts. We used a microarray technique to identify genes that were regulated in human osteoblasts and verified our findings using multiplex protein analysis and real-time RT-PCR. Recombinant ameloblastin was found to stimulate bone healing in vivo, and to enhance the proliferation of mesenchymal stem cells and osteoblasts, as well as the differentiation of osteoclast precursor cells in vitro. The most profound effect was on the regulation of genes related to immune responses as well as on the expression of cytokines and markers of bone cell differentiation, indicating that ameloblastin has an effect on mesenchymal cell differentiation. A receptor has not yet been identified, but we found rAmbn to induce, directly and indirectly, signal transducer and activator of transcription (STAT) 1 and 2 and downstream factors in the interferon pathway.

Ameloblastin Fusion Protein Enhances Pulpal Healing and Dentin Formation in Porcine Teeth

Ameloblastin (Ambn, also named “amelin” or “sheathlin”) is a protein participating in enamel formation and mesenchymal-ectodermal interaction during early dentin formation in developing teeth. Experiments have demonstrated an association between Ambn expression and healing of acute pulp wounds. The purpose of this study was to investigate if local application of recombinant fusion Ambn (rAmbn) could influence reparative dentin formation in pulpotomized teeth. In this randomized, double-blinded study, pulpotomy was performed in 28 lower central incisors in 17 adult miniature pigs. Following the surgical procedure, the exposed pulp tissue was covered either with rAmbn or with calcium hydroxide. After 2, 4, or 8 weeks, the teeth were extracted and examined by histomorphometry and immunohistochemistry using antibodies against porcine ameloblastin, collagen type I, and dentin sialoprotein (DSP). In rAmbn-treated teeth, a substantial amount of newly formed reparative dentin was observed at the application site, completely bridging the pulpal wound. Dentin formation was also observed in calcium hydroxide-treated teeth; however, the amount of reparative dentin was significantly smaller (P < 0.001) than after rAmbn treatment. Immunohistochemistry confirmed that the new hard tissue formed was similar to dentin. This is the first time a direct link between ameloblastin and dentin formation has been made in vivo. The results suggest potential for rAmbn as a biologically active pulp-dressing agent for enhanced pulpal wound healing and reparative dentin formation after pulpotomy procedures.

Effect of piezocision on root resorption associated with orthodontic force: A microcomputed tomography study

Introduction The purpose of this study was to investigate the effect of piezocision on orthodontically induced inflammatory root resorption. Methods Fourteen patients were included in this split‐mouth study; 1 side was assigned to piezocision, and the other side served as the control. Vertical corticotomy cuts of 4 to 5 mm in length were performed on either side of each piezocision premolar, and 150‐g buccal tipping forces were applied to the premolars. After 4 weeks, the maxillary first premolars were extracted and scanned with microcomputed tomography. Results There was a significantly greater total amount of root resorption seen on the piezocision sides when compared with the control sides (P = 0.029). The piezocision procedure resulted in a 44% average increase in root resorption. In 5 patients, there was noticeable piezocision‐related iatrogenic root damage. When that was combined with the orthodontic root resorption found on the piezocision‐treated teeth, there was a statistically significant 110% average increase in volumetric root loss when compared with the control side (P = 0.005). Conclusions The piezocision procedure that initiates the regional acceleratory phenomenon may increase the iatrogenic root resorption when used in conjunction with orthodontic forces. Piezocision applied close to the roots may cause iatrogenic damage to the neighboring roots and should be used carefully. HighlightsPiezocision might cause increased root resorption when used with orthodontic forces.Piezocision‐related iatrogenic root damage occurred in 5 of 14 participants.Piezocision should not be used in areas of close root proximity.

The efficacy of air polishing devices in supportive periodontal therapy: A systematic review and meta-analysis.

OBJECTIVE This systematic review analyzes existing literature on the clinical efficacy of air polishing devices (APDs), discussing the evidence-based data available for justifying their use as an alternative to conventional periodontal debridement in supportive periodontal therapy. The main objective of the review was to assess whether APD was as equally efficient or superior in obtaining successful treatment outcomes when compared with conventional methods. DATA SOURCES Following PRISMA guidelines, a systematic literature search of articles in English, up to December 2016, was conducted using PubMed, Cochrane, and Medline. Relevant articles were selected based on specific criteria. Seven studies were selected for the final assessment. One more study was added after a manual search of the literature. Due to considerable heterogeneity in study designs and outcome variables measured, only clinical parameters (probing depth, bleeding on probing, and clinical attachment level) were selected for meta-analysis. CONCLUSION The studies selected for this systematic review provide some evidence that APDs as monotherapy could be an alternative to conventional debridement of single- and multi-rooted teeth with no furcation involvement, during supportive periodontal therapy. Comparing clinical and microbiologic outcomes, APDs seem to be as effective as conventional treatments. The primary advantage for the use of APDs in supportive periodontal therapy seems to be their ability to efficiently remove biofilm, without causing damage to the periodontal soft tissues or tooth and root structure. There may also be an advantage regarding patient comfort and time consumed.

Automatic detection of periodontitis using intra-oral images

Periodontitis is a chronic inflammatory disease of the supportive tissues and bone surrounding the teeth. In severe cases, it can consequently lead to tooth loss. This disease is most prevalent in rural and remote communities where regular dental visits are limited. Hence, theres a need for a periodontal screening tool for use by allied health professionals outside of dental clinics to detect periodontitis for early referral and intervention. In this paper two algorithms have been proposed and applied on two independently collected datasets in Germany and Australia with 20 and 24 participating subjects respectively; in the first algorithm, intra-oral images of before periodontitis treatment have been considered as diseased subjects and the images of after treatment have been considered as healthy subjects. Using the histogram of pixel intensity as our classification feature, the healthy and diseased subjects have been classified with an accuracy of 66.7%. In the second algorithm, using the difference between the histograms as our classification features, images of “before” and “after” treatment have been classified with an accuracy of 91.6%. If used in a smart phone application, the first algorithm can help people with limited access to dental clinics to be screened for periodontitis by allied health professionals in any healthcare setting. The second algorithm may be useful in helping non-dental personnel to monitor the progress of periodontal treatment.