Optimal effective concentration combinations (OPECCs) for binary application of membrane-targeting antiseptics and TMPyP-mediated antimicrobial photodynamic therapy.

The widespread occurrence of multi-resistant bacteria is a health problem of global dimension. Infections caused by multi-resistant pathogens are difficult to treat and often associated with high mortality. Therefore, new treatment strategies are of interest, such as the use of differently acting antibacterial concepts. One of these new concepts is the use of antiseptics in combination with the antibacterial photodynamic therapy (aPDT). Currently, no method has yet been established as a standard procedure for investigating combined effects and evaluating them in a generally valid and unambiguous manner. The focus of this study was on how cationic antiseptics benzalkonium chloride (BAC) and chlorhexidine digluconate (CHX) behave in a combined application with aPDT using the photosensitizer TMPyP. For this purpose, BAC and CHX were applied in combination with the aPDT using TMPyP in non-lethal concentrations to the three bacteria Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis. The results of the combination experiments with sublethal concentrations of BAC or CHX with the aPDT showed that the binary application had a lethal effect. Irrespective of the bacteria, the reduction in concentrations in OPECC, compared to individual concentrations, was more than 50% for TMPyP, 23-40% for BAC, and 18-43% for CHX. Furthermore, the optimal effective concentration combinations (OPECCs) could be determined. The latter showed that the combined application allowed the reduction of both concentrations compared to the single application.

Stress response in Escherichia coli following sublethal phenalene-1-one mediated antimicrobial photodynamic therapy: an RNA-Seq study.

Since the molecular mechanisms behind adaptation and the bacterial stress response toward antimicrobial photodynamic therapy (aPDT) are not entirely clear yet, the aim of the present study was to investigate the transcriptomic stress response in Escherichia coli after sublethal treatment with aPDT using RNA sequencing (RNA-Seq). Planktonic cultures of stationary phase E. coli were treated with aPDT using a sublethal dose of the photosensitizer SAPYR. After treatment, RNA was extracted, and RNA-Seq was performed on the Illumina NextSeq 500. Differentially expressed genes were analyzed and validated by qRT-PCR. Furthermore, expression of specific stress response proteins was investigated using Western blot analysis.The analysis of the differential gene expression following pathway enrichment analysis revealed a considerable number of genes and pathways significantly up- or down-regulated in E. coli after sublethal treatment with aPDT. Expression of 1018 genes was up-regulated and of 648 genes was down-regulated after sublethal treatment with aPDT as compared to irradiated controls. Analysis of differentially expressed genes and significantly de-regulated pathways showed regulation of genes involved in oxidative stress response and bacterial membrane damage. In conclusion, the results show a transcriptomic stress response in E. coli upon exposure to aPDT using SAPYR and give an insight into potential molecular mechanisms that may result in development of adaptation.

Shear Bond Strength of Adhesives Placed following Selective Removal of Red-Fluorescing Carious Dentine in vitro.

Red-fluorescing dentine indicates bacterial contamination [Caries Res 2002; 36: 315-319]. We investigated effect of removal of red fluorescent dentine caries on shear bond strength and fracture mode of 4 adhesive approaches. Sixty-five carious teeth and 50 noncarious controls were distributed into 4 groups: Clearfil™ self-etch (CSE), OptiBond™ FL total etch (OTE), Scotchbond™ Universal total etch (STE) and self-etch (SSE). Samples were excited at 405 nm and viewed through 530 nm filter. Carious samples were ground flat exposing strongly red-fluorescing (StrongRF) dentine, on which a composite cylinder was placed, using one of 4 adhesives. After 22 h in water, shear bond strength and fracture mode were analysed. StrongRF was removed; composite cylinders were placed on weakly red-fluorescing (WeakRF) dentine and tested as described above. Finally, red-fluorescing dentine was removed, and composite cylinders were placed on non-fluorescing (NonRF) dentine and tested. Composites were placed at 3 corresponding heights in controls. After 22 h in water, shear bond strength testing and fracture mode analysis were performed. Differences were tested using Mann-Whitney or Wilcoxon tests (p ≤0.05). Median (Q1, Q3) shear bond strength on StrongRF was SSE 14.4 (9.2, 18.2) MPa >CSE 10.2 (6.4, 17.3) MPa >STE 9.1 (6.9, 11.2) MPa >OTE 6.8 (4.0, 10.8) MPa. Shear bond strength increased statistically significantly for all adhesives on WeakRF: SSE 19.8 (13.6, 24.3) MPa >STE 19.5 (12.7, 23.1) MPa >CSE 17.5 (12.0, 22.5) MPa >OTE 15.8 (11.9, 20.9) MPa. Only STE 25.6 (22.4, 29.1) MPa and CSE 22.1 (17.6, 24.6) MPa were significantly different on NonRF compared to WeakRF. For controls tested at corresponding depths, superficial shear bond strength was OTE 18.7 (16.0, 22.1) MPa >STE 18.4 (12.0, 25.9) MPa >CSE 18.1 (12.7, 20.7) MPa >SSE 13.0 (9.6, 17.8) MPa. This was significantly higher compared to StrongRF except for SSE. Central shear bond strength was not significantly different to WeakRF, deep shear bond strength was significantly lower for SSE and CSE but higher for OTE compared to carious. Conclusion: StrongRF dentine should be removed for higher shear bond strength, but WeakRF dentine can often be preserved without compromising adhesive bond strength.

Randomized controlled clinical trial on the efficacy of a novel antimicrobial chewing gum in reducing plaque and gingivitis in adolescent orthodontic patients.

OBJECTIVES: Chewing gums containing antiseptics or other antimicrobial substances may be effective in reducing plaque and gingivitis. Therefore, the aim of this randomized placebo-controlled clinical trial was to investigate the efficacy of a novel antimicrobial chewing gum containing essential oils (cinnamon, lemon, peppermint) and extracts on reduction of dental plaque and gingivitis as well as on oral health-related quality of life (OHRQoL) in adolescent orthodontic patients. MATERIALS: 52 patients (11-22 years of age) were randomly assigned to use a test chewing gum (COVIDGUM, Clevergum) or a commercially available control chewing gum over a period of 10 days. Approximal plaque index (API), papillary bleeding index (PBI) and an OHRQoL questionnaire for children (COHIP-G19) were assessed at baseline (BL), after 10 days (10d) and 30 days (30d). In addition, oral health and oral hygiene related questions of the COHIP-G19 questionnaire were evaluated separately in subscales at each timepoint. Data were analyzed using non-parametrical statistical procedures (α = 0.05). RESULTS: API and PBI decreased significantly over time from BL to 10d and from BL to 30d in both groups, without significant differences between the groups. In both groups, the COHIP-G19 score, oral health subscale and oral hygiene subscale decreased significantly over time. Regarding the oral hygiene subscale, the test group showed significantly better scores at 30d (p = 0.011). CONCLUSION: Both chewing gums performed similarly effective in terms of reducing plaque accumulation and gingival inflammation and improving OHRQoL. CLINICAL RELEVANCE: Chewing gums without antimicrobial ingredients may be sufficient to decrease plaque accumulation and gingival inflammation.

General diseases and medications in 687 patients reporting on adverse effects from dental materials.

OBJECTIVES: Examination of patients claiming adverse effects from dental materials can be very challenging. Particularly, systemic aspects must be considered besides dental and orofacial diseases and allergies. Therefore, the aim of this study was to investigate a cohort of 687 patients reporting on adverse effects from dental materials focusing on findings related to known general diseases or conditions or medication-related findings with relevance to their subjective complaints. METHODS: Six hundred eighty-seven patients visiting a specialized consultation on claimed adverse effects from dental materials were retrospectively investigated for their subjective complaints, findings related to known general diseases or conditions, medication-related findings, dental and orofacial findings, or allergies with relevance to their subjective complaints. RESULTS: The most frequent subjective complaints were burning mouth (44.1%), taste disorders (28.5%), and dry mouth (23.7%). In 58.4% of the patients, dental and orofacial findings relevant to their complaints could be found. Findings related to known general diseases or conditions or medication-related findings were found in 28.7% or 21.0% of the patients, respectively. Regarding medications, findings related to antihypertensives (10.0%) and psychotropic drugs (5.7%) were found most frequently. Relevant diagnosed allergies toward dental materials were found in 11.9%, hyposalivation in 9.6% of the patients. In 15.1% of the patients, no objectifiable causes for the expressed complaints could be found. CONCLUSIONS: For patients complaining of adverse effects from dental materials, findings related to known general diseases or conditions and medications should be given particular consideration, while still in some patients, no objectifiable causes for their complaints can be found. CLINICAL RELEVANCE: For patients complaining about adverse effects from dental materials, specialized consultations and close collaboration with experts from other medical fields are eligible.

Membrane damage as mechanism of photodynamic inactivation using Methylene blue and TMPyP in Escherichia coli and Staphylococcus aureus.

The worldwide threat of antibiotic resistance requires alternative strategies to fight bacterial infections. A promising approach to support conventional antibiotic therapy is the antimicrobial photodynamic inactivation (aPDI). The aim of this work was to show further insights into the antimicrobial photodynamic principle using two photosensitizers (PS) of different chemical classes, Methylene Blue (MB) and TMPyP, and the organisms Escherichia coli and Staphylococcus aureus as Gram-negative and Gram-positive representatives. Planktonic cultures of both species were cultured under aerobic conditions for 24 h followed by treatment with MB or TMPyP at various concentrations for an incubation period of 10 min and subsequent irradiation for 10 min. Ability to replicate was evaluated by CFU assay. Accumulation of PS was measured using a spectrophotometer. The cytoplasmic membrane integrity was investigated by flow cytometry using SYBR Green and propidium iodide. In experiments on the replication ability of bacteria after photodynamic treatment with TMPyP or MB, a killing rate of 5 log10 steps of the bacteria was achieved. Concentration-dependent accumulation of both PS was shown by spectrophotometric measurements whereby a higher accumulation of TMPyP and less accumulation of MB was found for S. aureus as compared to E. coli. For the first time, a membrane-damaging effect of TMPyP and MB in both bacterial strains could be shown using flow cytometry analyses. Furthermore, we found that reduction of the replication ability occurs with lower concentrations than needed for membrane damage upon MB suggesting that membrane damage is not the only mechanism of aPDI using MB.

Impact of access cavity cleaning on the seal of postendodontic composite restorations in vitro.

AIM: The aim of the study was to investigate the influence of cavity cleaning and conditioning on marginal integrity of directly placed post-endodontic composite class-I-restorations in vitro. METHODOLOGY: A total of 168 fully intact teeth without caries or fillings received pre-endodontic composite restorations (class-II) after their extraction. Occlusal endodontic access-cavities were prepared, and root canals were instrumented and filled with gutta-percha and an epoxy resin-based sealer. Prior to post-endodontic class-I-restoration, access cavities were completely contaminated with sealer, cleaned with alcohol and pre-treated as follows: cleaner only (alcohol), glycine-polishing, Al2 O3 sandblasting, carbide bur (immediate as well as delayed restoration). A positive control (not contaminated with sealer and adhesive used) and negative control (cleaner used but no adhesive) were established. Half of the teeth from each group were subjected to thermocycling and mechanical loading (TCML). Marginal integrity of post-endodontic restoration was evaluated in oro-vestibular or mesio-distal sections after AgNO3 dye penetration (DP) by standardized photomacroscopic imaging and expressed in per cent of margin length along all segments and separately for enamel, dentine and composite, respectively. Results were analysed non-parametrically (α = .05). RESULTS: No restorations or teeth fractured or debonded completely. Without TCML, the median DP of all segments was significantly higher for the negative control compared with all other groups in oro-vestibular cutting direction (53%; p = .002) and in mesio-distal cutting direction (51%; p ≤ .041). The other groups without TCML revealed 16%-24% DP (oro-vestibular) and 12%-24% DP (mesio-distal). With TCML, the median DP in oro-vestibular cutting direction for all segments ranged between 48% and 62% for all groups, a significant difference was only observed between glycine-polishing and carbide bur (p = .041). In mesio-distal cutting direction, the median DP in negative control was 69% with TCML and significantly higher compared with all other groups (p = .002). For all other groups, the median DP of all segments ranged between 28% and 40% with TCML without significant differences. Error rates method (k = 7) revealed a significant influence of TCML in general on penetration of all segments in both oro-vestibular and mesio-distal cutting directions. CONCLUSION: Additional access cavity pre-treatment after alcohol cleaning did not improve the marginal integrity of post-endodontic composite restorations. Thorough cleaning of the access cavity with alcohol seems to assure an acceptable marginal integrity to the tooth and restorative composite.

One-year results of a novel self-adhesive bulk-fill restorative and a conventional bulk-fill composite in class II cavities-a randomized clinical split-mouth study.

OBJECTIVES: In the context of the phase-down of amalgam, development of easily applicable, permanent restorative materials is of high clinical interest. Aim of this study was to evaluate the clinical performance of a novel, tooth-colored, self-adhesive bulk-fill restorative (SABF, 3M Oral Care) and a conventional bulk-fill composite (Filtek One, 3M Oral Care; FOBF) for restoring class II cavities. The null-hypothesis tested was that both materials perform similar regarding clinical performance. MATERIALS AND METHODS: In this randomized split-mouth study, 30 patients received one SABF and one FOBF restoration each. Scotchbond Universal (3M Oral Care) was used as adhesive for FOBF (self-etch mode), while SABF was applied directly without adhesive. Restorations were evaluated by two blinded examiners at baseline, 6 months and 12 months employing FDI criteria. Non-parametric statistical analyses and χ2-tests (α = 0.05) were applied. RESULTS: Thirty patients (60 restorations) were available for the 6- and 12-month recalls exhibiting 100% restoration survival. All restorations revealed clinically acceptable FDI scores at all time points and for all criteria. Only regarding esthetic properties, FOBF performed significantly better than SABF regarding surface lustre (A1) and color match and translucency (A3) at all time points and marginal staining (A2b) at 12 months. CONCLUSIONS: The null-hypothesis could not be rejected. Both materials performed similarly regarding clinical performance within the first year of clinical service. SABF exhibited slightly inferior, but clinically fully acceptable esthetic properties as compared to FOBF. CLINICAL RELEVANCE: Within the limitations of this study, the self-adhesive bulk-fill restorative showed promising results and may be recommended for clinical use.

Influence of autoclave sterilization on bond strength between zirconia frameworks and Ti-base abutments using different resin cements.

STATEMENT OF PROBLEM: Zirconia frameworks luted on titanium bases have been increasingly used for implant-supported prostheses. A reliable bond strength between the framework and the base is a key factor for clinical success. Autoclaving of the abutments before clinical use has been strongly suggested since abutments have been classified as class II medical devices. However, comprehensive knowledge on the susceptibility of the composite resin luting agents' adhesion to autoclave sterilization is lacking. PURPOSE: The purpose of this in vitro study was to assess the bond strength between zirconia frameworks and titanium bases using different composite resin luting agents with and without thermocycling and autoclaving. MATERIAL AND METHODS: A total of 200 titanium implants were copy milled and 200 zirconia frameworks (Cercon) were generated by computer-aided design and computer-aided manufacturing (CAD-CAM). They were adhesively luted using Multilink Hybrid Abutment (MHA), Panavia F2.0 (PF2), Panavia V5 (PV5), RelyX Ultimate (RXU), or Panavia SA Cement Plus (PSA). Autoclaving (134 °C, 0.21 MPa, 5.5 minutes) and thermocycling (3000 cycles, 5 °C and 55 °C) were performed. The push out bond strength was tested at a crosshead speed of 1 mm/min. Statistical analysis was performed by using the Mann-Whitney U test (α=.05) and the family-wise error rate method. RESULTS: Median bond strength values (interquartile range) after autoclaving and thermocycling ranged between 33.7 (26.7-41.8) MPa for MHA and 7.6 (7.3-8.5) MPa for PF2. MHA showed significantly higher values than all other composite resin luting agents regardless of thermocycling or autoclaving. PV5 showed equal or higher bond strength values than PSA, RXU, and PF2. Autoclaving had no significant influence on MHA or PV5, but had a negative effect on PSA and RXU. The family-wise error rate method showed that both the composite resin luting agent and autoclave sterilization had an overall influence on bond strength. CONCLUSIONS: MHA and PV5 were not significantly influenced by autoclaving and showed acceptable bond strength values. Because MHA showed significantly higher bond strength than all other materials and is released for autoclaving by the manufacturer, it is the first choice for extraoral bonding of zirconia frameworks to titanium bases. Manufacturers who state the indication of extraoral cementation of frameworks on titanium bases should also indicate whether their composite luting material is affected by autoclaving.

Relapse in class II orthognathic surgery: a systematic review.

OBJECTIVES: Relapse after orthognathic surgery seems to depend on diverse factors. Proffit et al. postulated in 2007 a "hierarchy of stability" (Head Face Med 6:66, 2007), ranking posttreatment stability after various orthognathic procedures, but no systematically reviewed evidence was provided. Therefore, the aim of this review was to investigate the extent of class II relapse in orthognathic surgery of Angle class II patients depending on the surgical procedure used. MATERIALS AND METHODS: Seven databases were searched for randomized and controlled clinical trials to compare relapse in surgical procedures for Angle class II patients. After duplicate study selection, data extraction and risk of bias assessment were performed with the ROBINS-I tool as well as data synthesis by frequency distribution, followed by assessment of the quality of evidence with GRADE. RESULTS: Four non-randomized cohort-studies with a total of 132 patients were included. Bimaxillary procedures as well mandibular advancement procedures proved to be highly stable. Single jaw interventions at the maxilla achieved mostly stable results at sagittal dimension and problematic stability in the vertical dimension. However, there were only limited data available with low quality of evidence. CONCLUSIONS: Limited existing evidence of low quality partly support the postulated hierarchy of stability of Proffit et al. (Head Face Med 6:66, 2007) and indicates that a surgical correction of class II dysgnathia with bimaxillary procedures and mandibular advancement seems to be highly stable. However, additional studies are needed to address the relation between relapse and surgical orthognathic intervention. Trial registration PROSPERO 2019 CRD42019144873.

Colonization of Helicobacter pylori in the oral cavity - an endless controversy?

Helicobacter pylori is associated with chronic gastritis, gastric or duodenal ulcers, and gastric cancer. Since the oral cavity is the entry port and the first component of the gastrointestinal system, the oral cavity has been discussed as a potential reservoir of H. pylori. Accordingly, a potential oral-oral transmission route of H. pylori raises the question concerning whether close contact such as kissing or sharing a meal can cause the transmission of H. pylori. Therefore, this topic has been investigated in many studies, applying different techniques for detection of H. pylori from oral samples, i.e. molecular techniques, immunological or biochemical methods and traditional culture techniques. While molecular, immunological or biochemical methods usually yield high detection rates, there is no definitive evidence that H. pylori has ever been isolated from the oral cavity. The specificity of those methods may be limited due to potential cross-reactivity, especially with H. pylori-like microorganisms such as Campylobacter spp. Furthermore, the influence of gastroesophageal reflux has not been investigated so far. This review aims to summarize and critically discuss previous studies investigating the potential colonization of H. pylori in the oral cavity and suggest novel research directions for targeting this critical research question.

Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon repeated exposure.

OBJECTIVES: The aims of this study were to investigate the antimicrobial efficacy of antiseptics in saliva-derived microcosm biofilms, and to examine phenotypic adaption of bacteria upon repeated exposure to sub-inhibitory antiseptic concentrations. METHODS: Saliva-derived biofilms were formed mimicking caries- or gingivitis-associated conditions, respectively. Microbial compositions were analyzed by semiconductor-based 16S rRNA sequencing. Biofilms were treated with CHX, CPC, BAC, ALX, and DQC for 1 or 10 min, and colony forming units (CFU) were evaluated. Phenotypic adaptation of six selected bacterial reference strains toward CHX, CPC, and BAC was assessed by measuring minimum inhibitory concentrations (MICs) over 10 passages of sub-inhibitory exposure. Protein expression profiles were investigated by SDS-PAGE. RESULTS: Both biofilms showed outgrowth of streptococci and Veillonella spp., while gingivitis biofilms also showed increased relative abundances of Actinomyces, Granulicatella, and Gemella spp. Antiseptic treatment for 1 min led to no relevant CFU-reductions despite for CPC. When treated for 10 min, CPC was most effective followed by BAC, ALX, CHX, and DQC. Stable adaptations with up to fourfold MIC increases were found in E. coli toward all tested antiseptics, in E. faecalis toward CHX and BAC, and in S. aureus toward CPC. Adapted E. coli strains showed different protein expression as compared with the wildtype strain. CONCLUSION: Antiseptics showed limited antimicrobial efficacy toward mature biofilms when applied for clinically relevant treatment periods. Bacteria showed phenotypic adaptation upon repeated sub-inhibitory exposure. CLINICAL RELEVANCE: Clinicians should be aware that wide-spread use of antiseptics may pose the risk of inducing resistances in oral bacteria.

Cetylpyridinium Chloride: Mechanism of Action, Antimicrobial Efficacy in Biofilms, and Potential Risks of Resistance.

Antimicrobial resistance is a serious issue for public health care all over the world. While resistance toward antibiotics has attracted strong interest among researchers and the general public over the last 2 decades, the directly related problem of resistance toward antiseptics and biocides has been somewhat left untended. In the field of dentistry, antiseptics are routinely used in professional care, but they are also included in lots of oral care products such as mouthwashes or dentifrices, which are easily available for consumers over-the-counter. Despite this fact, there is little awareness among the dental community about potential risks of the widespread, unreflected, and potentially even needless use of antiseptics in oral care. Cetylpyridinium chloride (CPC), a quaternary ammonium compound, which was first described in 1939, is one of the most commonly used antiseptics in oral care products and included in a wide range of over-the-counter products such as mouthwashes and dentifrices. The aim of the present review is to summarize the current literature on CPC, particularly focusing on its mechanism of action, its antimicrobial efficacy toward biofilms, and on potential risks of resistance toward this antiseptic as well as underlying mechanisms. Furthermore, this work aims to raise awareness among the dental community about the risk of resistance toward antiseptics in general.

Tooth survival and clinical outcomes up to 26 years after guided tissue regeneration therapy in deep intra-bony defects: Follow-up investigation of three randomized clinical trials.

AIM: To investigate tooth survival and clinical long-term outcomes up to 26 years following guided tissue regeneration (GTR) therapy in deep intra-bony defects. METHODS: Patients from three prospective clinical split-mouth studies, which investigated the outcomes of GTR therapy, were re-evaluated 21-26 years after surgery independent of the membrane type used, and tooth survival was assessed according to several site-specific and patient-related factors. RESULTS: About 50 patients contributing 102 defects were available for this long-term follow-up. After up to 26 years (median 23.3 years), 52.9% of the teeth were still in situ. The median survival of the extracted teeth was 13.8 years. Patients with diabetes mellitus and/or smoking history lost significantly more teeth in the long term. Compared to the 1-year situation, there was no new median CAL loss after up to 26 years in the teeth which were still in situ. CONCLUSIONS: Within the limitations of this study, our data show that more than 50% of the initially seriously diseased teeth were still in situ up to 26 years following GTR therapy despite an overall limited adherence to SPT. In the majority of these teeth, the CAL gain 1 year after GTR could be maintained over this long period.

Influence of selective caries excavation on marginal penetration of class II composite restorations in vitro.

Selective caries excavation may support pulp preservation. This in vitro study investigated the influence of selective removal of demineralized dentin on marginal integrity of composite restorations as determined by dye penetration. Dentinal caries-like lesions were produced in the approximal surfaces of 40 extracted human molars (ethylenediaminetetraacetate, 0.5 M, 96 h). The following test procedures were established: complete excavation, selective excavation, and caries-free control. Two class II cavities with enamel at the cervical margins were prepared per tooth and demineralization volume was determined by micro-computed tomography for the purpose of a stratified distribution to receive complete excavation or selective excavation. After complete or selective excavation (30 cavities each), adhesive composite restorations were placed. Cavities without demineralized dentin (20 cavities) served as control. The marginal integrity of restorations was evaluated by dye penetration with and without thermocycling or mechanical loading. Results were analyzed by non-parametrical statistical tests (Mann-Whitney U Test) with an α = 0.05 level of significance. Dye penetration did not differ significantly among completely excavated, selectively excavated, or undemineralized teeth, but was increased by thermocycling and mechanical loading in all experimental groups. Selective caries removal did not increase marginal penetration in class II restorations. The presence of remaining demineralized dentin surrounded by sound dentin did not impair marginal integrity of restorations with margins placed in sound enamel.

Longevity of posterior composite and compomer restorations in children placed under different types of anesthesia: a retrospective 5-year study.

OBJECTIVES: The aims of this study were (i) to assess cumulative survival rates of class II resin-based composite and compomer restorations in primary molars with a 5-year observation period and (ii) to analyze the influence of different types of anesthesia and different localizations of the restorations in the teeth. METHODS: Patient charts of a private practice for pediatric dentistry were screened for class II resin-based composite (Spectrum TPH3) and compomer (Dyract Posterior; both Dentsply DeTrey) restorations in primary molars with a 5-year observation period used with Adper Prompt L-Pop (3M-ESPE). One restoration per patient (age ≤ 6 years at placement) was randomly selected. RESULTS: Two hundred sixty restorations were included (43% resin-based composites, 57% compomers). After 5 years, cumulative survival rates were 43% for resin-based composite and 49% for compomer restorations with no statistically significant differences. There was a tendency for higher survival rates for restorations placed under N2O inhalation sedation or general anesthesia. Distal-occlusal compomer restorations showed significantly lower survival rates (p = 0.003) as compared to mesial-occlusal compomer restorations. CONCLUSION: Within the limitations of the study, we conclude that type of restorative material as well as the type of anesthesia do not influence restoration survival rates, although restorations placed in patients receiving N2O inhalation sedation or general anesthesia tend to perform better as compared with patients receiving no anesthesia or only local infiltration. CLINICAL RELEVANCE: Resin-based composite and compomer restorations show similar survival rates of more than 43% (annual failure rates less than 11.5%) after 5 years for restoration of primary molars.

Ca2+ release and buffering effects of synthetic hydroxyapatite following bacterial acid challenge.

BACKGROUND: Synthetic particulate hydroxyapatite (HAP; Ca5(PO4)3(OH)) is used as ingredient in oral care products but its effects on cariogenic biofilms are not clear yet. The primary mode of action of HAP may be acting as a calcium phosphate reservoir when deposited in oral biofilms and release Ca2+ and (hydrogen) phosphate ions upon bacterial acid challenge. The aim of this in vitro study was to test this hypothesis by investigating release of Ca2+ ions and potential buffering effects from HAP upon bacterial acid challenge in planktonic cultures and biofilms of Streptococcus mutans. METHODS: Planktonic cultures of S. mutans were grown in BHI broth with 1% sucrose or with additional 5% HAP or 5% silica for up to 48 h. Separately, biofilms of S. mutans were grown in BHI for 72 h in total. After 24 h of this biofilm culture, either BHI alone or BHI with additional 0.5% HAP or 0.5% silica was added. After 48 h, BHI with 1% sucrose was added to allow bacterial acid formation. Ca2+ release was determined colorimetrically and pH measurements were performed using a pH electrode. For statistical analysis, non-parametrical procedures were applied (n ≥ 10; Mann-Whitney U test; α = 0.05). RESULTS: Relevant release of Ca2+ was only evident in planktonic cultures or biofilms with HAP but not in both other groups (p ≤ 0.001). In suspended biofilms with HAP, median pH was 4.77 after 72 h and about 0.5 pH units higher as compared to both other groups (4.28 or 4.32, respectively; p ≤ 0.001). CONCLUSIONS: Under the tested conditions, synthetic HAP releases Ca2+ ions upon bacterial acid challenge and may also show some buffering capacity but further studies are needed to investigate whether the concentrations tested here can also be reached clinically in dental biofilms.

Influence of autogenous platelet concentrate on combined GTR/graft therapy in intra-bony defects: A 13-year follow-up of a randomized controlled clinical split-mouth study.

AIM: To investigate the clinical long-term outcomes 13 years following guided tissue regeneration (GTR) in deep intra-bony defects with and without additional application of autogenous platelet concentrate (APC). METHODS: In 25 patients, two deep contra-lateral intra-bony defects were treated according to GTR using ß-TCP and bio-resorbable membranes. In test defects, APC was applied additionally. After 13 years, clinical healing results were assessed and compared to results at baseline and after 1 year. Furthermore, a tooth survival analysis was carried out. RESULTS: After 13 years, 22 patients were available for tooth survival analysis showing 81.8% of test and 86.4% of control teeth still in situ. Based on the 15 patients still available for split-mouth analysis, median CAL was 10.0 mm in test and 12.0 mm in control sites at baseline. After 1 year, both groups revealed significant CAL gains of 5.0 mm, followed by a new CAL loss of 1.0 mm in the following 12 years. There were no significant differences between test and control sites. CONCLUSION: Within the limits of this study, the data show that most of the CAL gain following GTR can be maintained over 13 years. The additional use of APC had no positive influence on the long-term stability.

Phenalen-1-One-Mediated Antimicrobial Photodynamic Therapy and Chlorhexidine Applied to a Novel Caries Biofilm Model.

Antimicrobial photodynamic therapy (aPDT) may be useful as a supportive antimicrobial measure for caries-active subjects. In this study, the antimicrobial efficacy of aPDT with a phenalen-1-one photosensitizer was evaluated in a novel in vitro biofilm model comprising Actinomyces naeslundii, Actinomyces odontolyticus, and Streptococcus mutans and was compared to chlorhexidine. The proposed biofilm model allows high-throughput screening for antimicrobial efficacy while exhibiting a differentiated response to different antimicrobial approaches. While chlorhexidine 0.2% showed a reduction of ≈4 log10 for all species, aPDT led to a more pronounced reduction of S. mutans (2.8 log10) than of Actinomyces spp. (1.2 or 1.3 log10). A similar effect was also observed in monospecies biofilms. Therefore, aPDT may be more effective against S. mutans than against Actinomyces spp. when in biofilms, and this antimicrobial approach merits further investigations.

Effect of bleaching agent extracts on murine macrophages.

OBJECTIVES: The aim of this study was to evaluate the cytotoxicity and the influence of bleaching agents on immunologically cell surface antigens of murine macrophages in vitro. MATERIALS AND METHODS: RAW 264.7 cells were exposed to bleaching gel extracts (40% hydrogen peroxide or 20% carbamide peroxide) and different H2O2 concentrations after 1 and 24-h exposure periods and 1-h exposure and 23-h recovery. Tests were performed with and without N-acetyl cysteine (NAC) and buthionine sulfoximine (BSO). Cell viability was determined by MTT assay. The expression of surface markers CD14, CD40, and CD54 with and without LPS stimulation was detected by flow cytometry, while the production of TNF-α was measured by ELISA. Statistical analysis was performed using the Mann-Whitney U test (α = 0.05). RESULTS: Extracts of bleaching agents were cytotoxic for cells after a 1-h exposure; cells could not recover after 24 h. This effect can be mitigated by the antioxidant NAC and increased by BSO, an inhibitor of glutathione (GSH) synthesis. LPS stimulated expression of all surface markers and TNF-α production. Exposure to bleaching agent extracts and H2O2 leads to a reduction of TNF-α, CD14, and CD40 expression, while the expression of CD54 was upregulated at non-cytotoxic concentrations. Whereas NAC reduced this effect, it was increased in the presence of BSO. CONCLUSIONS: Extracts of bleaching agents were irreversibly cytotoxic to macrophages after a 1-h exposure. Only the expression of CD54 was upregulated. The reactions are mediated by the non-enzymatic antioxidant GSH. CLINICAL RELEVANCE: The addition of an antioxidant can downregulate unfavorable effects of dental bleaching.