In Vitro Cytotoxic and Inflammatory Response of Gingival Fibroblasts and Oral Mucosal Keratinocytes to 3D Printed Oral Devices.

The purpose of this study was to examine the biocompatibility of 3D printed materials used for additive manufacturing of rigid and flexible oral devices. Oral splints were produced and finished from six printable resins (pairs of rigid/flexible materials: KeySplint Hard [KR], KeySplint Soft [KF], V-Print Splint [VR], V-Print Splint Comfort [VF], NextDent Ortho Rigid [NR], NextDent Ortho Flex [NF]), and two types of PMMA blocks for subtractive manufacturing (Tizian Blank PMMA [TR], Tizian Flex Splint Comfort [TF]) as controls. The specimens were eluted in a cell culture medium for 7d. Human gingival fibroblasts (hGF-1) and human oral mucosal keratinocytes (hOK) were exposed to the eluates for 24 h. Cell viability, glutathione levels, apoptosis, necrosis, the cellular inflammatory response (IL-6 and PGE2 secretion), and cell morphology were assessed. All eluates led to a slight reduction of hGF-1 viability and intracellular glutathione levels. The strongest cytotoxic response of hGF-1 was observed with KF, NF, and NR eluates (p < 0.05 compared to unexposed cells). Viability, caspase-3/7 activity, necrosis levels, and IL-6/PGE2 secretion of hOK were barely affected by the materials. All materials showed an overall acceptable biocompatibility. hOK appeared to be more resilient to noxious agents than hGF-1 in vitro. There is insufficient evidence to generalize that flexible materials are more cytotoxic than rigid materials. From a biological point of view, 3D printing seems to be a viable alternative to milling for producing oral devices.

X-Linked Hypophosphatemia: Does Targeted Therapy Modify Dental Impairment?

X-linked hypophosphatemia is a rare, hereditary disorder that significant influences teeth and alveolar bone. The first clinical sign leading to the diagnosis of X-linked hypophosphatemia is often dental impairment with dental abscesses and dentin mineralization defects. Genetic analysis helped find the responsible gene and therefore opened up new ways of therapeutically managing X-linked hypophosphatemia. The human monoclonal antibody Burosumab represents a milestone in the targeted therapy of this hereditary disease by directly addressing its pathophysiology. Targeted therapy has been shown to improve skeletal impairment, pain, and phosphate metabolism. However, the influence of this new therapy on dental impairment has only been addressed in a few recent studies with varying results. Therefore, in this review, we aim to summarize the dental phenotype and analyze the different treatment modalities with a focus on dental impairment.

Recording the maxillomandibular relationship with the Aqualizer system prior to occlusal splint therapy for treating temporomandibular disorders: a randomized controlled trial.

Temporomandibular disorders (TMD) present a public health issue and are one of the most common musculoskeletal conditions causing chronic pain. This study compares the outcomes of occlusal splint therapy in patients with TMD following two different maxillomandibular relationship (MMR) registration techniques. 40 TMD patients were randomly allocated to MMR registration with the Aqualizer system (AQU) or with chin point guidance (CPG) prior to fabricating occlusal splints. TMD symptoms, subjective pain intensity, and quality of life (QoL) were recorded at baseline and after 3 and 6 months. The treatment led to an overall reduction of TMD symptoms in both groups (Conover test, p < 0.00001). TMJ sounds, TMJ pain with palpation and muscle pain with palpation subsided regardless of the type of MMR registration method used (Cohen's d > 0.8). AQU-based occlusal splints led to a better improvement of TMJ pain with maximum opening compared to CPG-based occlusal splints (Cohen's d = 0.9; CPG d = 0.13). In both groups, occlusal splint treatment had little to no effect on correcting lateral mandible deviation or improving restricted jaw opening. After 6 months occlusal splints in both groups had a large effect on improving subjective pain intensity (Cohen's d > 0.8), however, patients reported a higher QoL in the AQU group compared to the CPG group (Mann-Whitney-U-test, p < 0.05). The results of this study support the premise that occlusal splints are effective in relieving pain-related TMD symptoms. The Aqualizer can be considered for determining MMR in cases, where guided registration techniques are not possible.Trial registration: DRKS00031998.

Metastases of OSCC Based on Oral Lichen Ruber Planus.

Oral lichen ruber planus (OLP) is a poorly understood chronically inflammatory disease of the oral mucosa. Malignant transformation into oral squamous cell carcinoma (OSCC) is reported in between 1-2% of cases in the literature. After malignant transformation, surgical treatment-meaning tumor resection combined with neck dissection-is recommended. The recommended extent of treatment is controversial in the literature because this kind of OSCC is often a highly differentiated tumor with a lower risk for lymph nodal spreading. This study aims to overview 103 patients treated in our department due to OLP. The primary outcome parameter was the development of metastases in OLP patients compared to a group of OSCC patients without OLP and the comparison of survival in between both groups. Statistical analysis showed a significantly lower risk for patients with OSCC and with OLP for lymph nodal spreading (p = 0.013). Patients with OSCC and without OLP had a 4.76-higher risk for lymph nodal spreading. On the other hand, second metachronous tumor occurred more often in patients with OSCC and OLP. Overall, OSCC based on OLP occurs more often in female patients, is more highly differentiated and comes with a lower risk for metastases but has a higher risk for second metachronous tumors. Therefore, special attention should be paid to patients with OSCC based on OLP when planning adjuvant therapy and clinical follow-up. The indication for postoperative radiation should be made cautiously in this case, and clinical controls should be performed more closely due to the risk of recurrent disease or tumors at different locations.

Microleakage of Restorative Materials Used for Temporization of Endodontic Access Cavities.

A tight temporary seal applied to an access cavity is thought to improve endodontic outcomes. This study aims to assess the bacterial and glucose microleakage of different types and combinations of temporary restorations. Human-extracted incisors were instrumented, dressed with a calcium hydroxide paste, and sealed with Cavit W (CW), CW/Ketac Molar (CW/KM), CW/Smart Dentin Replacement (CW/SDR), Intermediate restorative material/KM (IRM/KM), or Clip F (CF). Standardized 3D-printed hollow test specimens were manufactured and temporized in the same manner. The specimens were examined for bacterial and glucose leakage for 28 days. Data were analyzed using a Kaplan-Meier survival analysis. CW/SDR and CF showed the least bacterial and glucose leakage over time. CW, CW/KM, and IRM/KM had similarly high levels of glucose leakage, but CW/KM and IRM/KM provided a tighter seal against bacterial penetration than CW. CW/SDR and CF should be considered for the sealing of access cavities of teeth previously restored with methacrylate-based materials.

Leaching components and initial biocompatibility of novel bioactive restorative materials.

OBJECTIVES: Bioactive restorative materials were developed on the premise that direct restorations should not only serve the purpose of reconstructing dental hard tissue defects but also exhibit biological features that prevent secondary caries development, without having adverse effects on the host cells. This study focuses on assessing the in vitro biocompatibility of two novel bioactive restorative materials. METHODS: Specimens of the bioactive restorative materials, Cention Forte (CF) and ACTIVA BioACTIVE RESTORATIVE (AB), a glass ionomer cement/glass hybrid (EQUIA Forte HT, EF) and an established nanohybrid composite (Venus Diamond, VD) were produced and finished. The specimens were eluted in water and methanol and the resulting eluates were analyzed via gas chromatography-mass spectrometry. hGF-1 cells were exposed to eluates prepared in cell culture medium. Cellular ATP levels, oxidized glutathione concentration, caspase-3/7 activity and the inflammatory response (IL-6 and PGE2 levels) were determined. Microscopic images were taken to examine the cell morphology. RESULTS: Methyl methacrylate and 2-Hydroxyethyl methacrylate were the main monomers detected in CF and AB eluates. All materials inhibited cell proliferation and led to significantly reduced ATP-levels. The cells exhibited a healthy morphology in the presence of CF and AB. Cells exposed to VD showed increased oxidized glutathione levels. Only EF led to enhanced caspase-3/7 activity. CF and AB caused IL-6 levels to increase, while EF and AB led to enhanced PGE2 levels. SIGNIFICANCE: CF and AB are promising materials from a biological point of view and seem to have improved bioactive properties compared to glass ionomer cements.

Surface properties and initial bacterial biofilm growth on 3D-printed oral appliances: a comparative in vitro study.

OBJECTIVES: To investigate the initial bacterial adhesion on 3D-printed splint materials in relation to their surface properties. MATERIALS AND METHODS: Specimens of five printable splint resins (SHERAprint-ortho plus UV, NextDent Ortho Rigid, LuxaPrint Ortho Plus, V-Print Splint, KeySplint Soft), one polymethylmethacrylate (PMMA) block for subtractive manufacturing (Astron CLEARsplint Disc), two conventional powder/liquid PMMA materials (FuturaGen, Astron CLEARsplint), and one polyethylene terephthalate glycol (PETG) thermoplastic sheet for vacuum forming (Erkodur Thermoforming Foil) were produced and finished. Surface roughness Ra was determined via contact profilometry. Surface morphology was examined under a scanning electron microscope. Multi-species bacterial biofilms were grown on entire splints. Total biofilm mass and viable bacterial counts (CFU/ml) within the biofilms were determined. Statistical analyses were performed with a one-way ANOVA, Tukey's post hoc test, and Pearson's test (p < 0.05). RESULTS: Astron CLEARsplint and KeySplint Soft specimens showed the highest surface roughness. The mean total biofilm mass on KeySplint Soft splints was higher compared to all other materials (p < 0.05). Colony-forming unit per milliliter on FuturaGen, Astron CLEARsplint, and KeySplint Soft splints was one log scale higher compared to all other materials. The other four printable resins displayed overall lower Ra, biofilm mass, and CFU/ml. A positive correlation was found between Ra and CFU/ml (r = 0.69, p = 0.04). CONCLUSIONS: The 3D-printed splints showed overall favorable results regarding surface roughness and bacterial adhesion. Thermoplastic materials seem to display a higher surface roughness, making them more susceptible to microbial adhesion. CLINICAL RELEVANCE: The development of caries and gingivitis in patients with oral appliances may be affected by the type of material.

Initial biocompatibility of novel resins for 3D printed fixed dental prostheses.

OBJECTIVES: Several materials for 3D printing of fixed dental prostheses (FDP) have been recently introduced. This study aims to evaluate the initial biocompatibility of novel printable resins for manufacturing temporary and permanent FDP. METHODS: Specimens of five printable resins (VarseoSmile Crown plus, NextDent C&B MFH, VarseoSmile Temp, Temp PRINT, P Pro Crown & Bridge), two types of resins for subtractive manufacturing (Tetric CAD, Telio CAD) and two types of resins with conventional curing processes (Tetric EvoCeram, Protemp 4) were produced and finished. Post-processing was strictly performed according to the manufacturer's protocol. Biocompatibility was evaluated by eluting specimens with cell culture medium and treating human gingival fibroblast cells with the eluates. A 72-hour continuous read cell viability assay measuring the reducing potential of the cells was performed. The cellular inflammatory response in terms of IL-6 and PGE2 levels was determined with specific ELISAs. Oxidative stress was determined by measuring oxidized glutathione concentrations after exposure to the resin eluates. A luminescence-based apoptosis assay was used to detect apoptosis. RESULTS: Tetric CAD and Telio CAD were slightly toxic. All other resins were moderately to severely cytotoxic. VarseoSmile Crown plus and P Pro Crown & Bridge significantly enhanced PGE2 levels. Higher concentrations of oxidized gluthatione were determined in the presence of Telio CAD, VarseoSmile Temp and P Pro Crown & Bridge. Tetric EvoCeram and Protemp 4 reduced intracellular gluthatione levels. All printable resins slightly induced apoptosis. SIGNIFICANCE: Further post-processing steps such as additional curing and washing may improve the biocompatibility of printable materials.

E-scooter-related craniomaxillofacial injuries compared with bicycle-related injuries - A retrospective study.

The aim of the study was to compare incidences and types of injury incurred between e-scooter crashes and bicycle crashes. This retrospective study included all victims of e-scooter crashes who were treated in the department of oral and maxillofacial surgery during a 12-months interval.A comparison was made with a cohort of patients who underwent bicycle crashes. Study parameters included type of fracture, soft tissue and dental affection, necessity of inpatient or outpatient treatment, the use of helmets, and the time of admission. In total, 400 patients were included. Of these 40 had suffered a crash on an e-scooter and 360 on a bicycle. Descriptive statistics showed a low helmet-wearing rate among cyclists (16.1% of recorded cases), with no helmet wearing recorded among e-scooter users. E-scooter-related crashes showed a higher rate of facial soft-tissue injuries (77.5%, p = 0.049) than among cyclists (61.7%), as well as a higher rate of dental injuries (27.5%, p = 0.017) compared with the bicycle cohort (13.3%). Facial fractures were also more common in the e-scooter cohort (45% vs 25.8%, p = 0.010). Admission was typically at the weekend - in the afternoon for the bicycle cohort and in the evening and at night for the e-scooter cohort. As a consequence of the fact that e-scooter riders seem to have an increased risk of facial injuries, it seems that an awareness campaign might be necessary to encourage helmet usage and to persuade intoxicated persons to use public transportation instead.

A comprehensive in vitro comparison of the biological and physicochemical properties of bioactive root canal sealers.

OBJECTIVES: To evaluate the biological and physicochemical features of bioactive root canal sealers. MATERIALS AND METHODS: Human periodontal ligament fibroblasts (hPDLF) and human osteoblasts (hOB) were exposed to eluates of three bioactive root canal sealers, GuttaFlow® bioseal (GF), BioRoot™ RCS (BR), and TotalFill® BC Sealer (TF), and the epoxy resin-based sealer AH plus® (AH). Cytotoxicity and cellular inflammatory response were evaluated. The osteogenic potential was examined using human mesenchymal stem cells (hMSC). Film thickness, flowability, and pH were assessed. Root canal treatment was performed on human extracted teeth to evaluate the sealers' tightness towards bacterial penetration. The antibacterial activity against common pathogens in primary root canal infections was tested. RESULTS: AH was severely cytotoxic to hPDLF and hOB (p < 0.001). The bioactive sealers were generally less cytotoxic. IL-6 levels in hPDLF were elevated in the presence of AH (p < 0.05). AH and GF suppressed IL-6 production in hOB (p < 0.05). AH and BR stimulated the PGE2 production in hPDLF and hOB (p < 0.05). BR was the only sealer that led to calcium deposits in hMSC (p < 0.05). TF and AH showed the lowest film thickness and the highest flowability. Bacterial tightness was best in teeth filled with AH and BR. All sealers showed similar antimicrobial activity, but the overall antimicrobial efficacy was moderate as the bacteria were reduced by just one log scale (p < 0.05). CONCLUSIONS: This study revealed favorable in vitro results regarding the biocompatibility of the bioactive root canal sealers. CLINICAL RELEVANCE: Bioactive root canal sealers may be a useful alternative to epoxy resin-based sealers.

Intranuclear cell uptake and toxicity of titanium dioxide and zirconia particles as well as bacterial adhesion on dental titanium- and zirconia-implants.

OBJECTIVE: Previous studies have shown that particles can be released from dental titanium (Ti)- and zirconia (ZrO2)-implants. Titanium dioxide (TiO2)- and ZrO2-particles were compared regarding their toxicity and intranuclear cell uptake as well as the adhesion of various anaerobic bacteria on Ti- and ZrO2-implants. METHODS: Cyto- and genotoxicity of TiO2-microparticles (TiO2-MPs) and TiO2-nanoparticles (TiO2-NPs) in periodontal ligament (PDL)-hTERT cells were determined with XTT test and DNA damage with comet assay. Particle sizes of TiO2- and ZrO2-particles were measured with scanning electron microscope. Intranuclear uptake in PDL-hTERT cells was determined with laser scanning confocal microscopy. Adhesions of relevant anaerobic mouth bacteria Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans on Ti- and ZrO2-implants were investigated by cultivation and counting bacterial colonies. RESULTS: Particle size measurements revealed that 99% of the TiO2-NPs had a size below 100 nm and 88% of the TiO2-MPs sizes were between 50 and 200 nm. Following EC50 values were found for particles (mg/l): 92 (TiO2-MPs) and 15 (TiO2-NPs). A significant increase in olive tail moment (OTM) was found for TiO2-NPs at a concentration of 1/10 EC50. TiO2- and ZrO2-NPs had a higher intranuclear cell uptake efficiency, compared to corresponding TiO2- and ZrO2-MPs. All investigated particles could be detected in cell nucleus. Adhesion of all investigated bacterial species was significantly higher on Ti-implants, compared to ZrO2-implants. CONCLUSION: Ti usually develops an oxide layer (TiO2). Particles released from Ti-implants should be TiO2-particles or Ti-particles coated with a TiO2-layer. Toxicity of released Ti-particles depends on their oxidation state and on their size (NP or MP). Particularly, NPs were more cyto- and genotoxic compared to the corresponding MPs. TiO2- and ZrO2-NPs showed a significant increase in the intranuclear cell uptake ratio at higher exposure concentration, compared to lower concentrations and consequently might lead to a higher potential of DNA damage. Adhesion of bacteria to ZrO2-implants is reduced, compared to Ti-implants. Therefore, ZrO2-implants might contribute to reduced biological complications (e.g. periimplantitis).

Special Teaching Formats during the COVID-19 Pandemic-A Survey with Implications for a Crisis-Proof Education.

Modern teaching formats have not been considered necessary during the COVID-19 pandemic with uncertain acceptance by students. The study's aim was to describe and evaluate all measures undertaken for theoretical and practical knowledge/skill transfer, which included objective structured practical examinations (OSPEs) covering a communication skills training. The students' performance in the OSPE as well as the theoretical knowledge level were assessed, of which the latter was compared with previous terms. In conservative dentistry and periodontology (4th and 5th year courses), theoretical teaching formats were provided online and completed by a multiple-choice test. Practical education continued without patients in small groups using the phantom-head, 3D printed teeth, and objective structured practical examinations (OSPEs) including communication skills training. Formats were evaluated by a questionnaire. The organization was rated as very good/good (88.6%), besides poor Internet connection (22.8%) and Zoom® (14.2%) causing problems. Lectures with audio were best approved (1.48), followed by practical videos (1.54), live stream lectures (1.81), treatment checklists (1.81), and virtual problem-based learning (2.1). Lectures such as .pdf files without audio, articles, or scripts were rated worse (2.15-2.30). Phantom-heads were considered the best substitute for patient treatment (59.5%), while additional methodical efforts for more realistic settings led to increased appraisal. However, students performed significantly worse in the multiple-choice test compared to the previous terms (p < 0.0001) and the OSPEs revealed deficits in the students' communication skills. In the future, permanent available lectures with audio and efforts toward realistic treatment settings in the case of suspended patient treatment will be pursued.

Targeting antibiotic tolerance in anaerobic biofilms associated with oral diseases: Human antimicrobial peptides LL-37 and lactoferricin enhance the antibiotic efficacy of amoxicillin, clindamycin and metronidazole.

Antimicrobial peptides are receiving increasing attention as potential therapeutic agents for treating biofilm-related infections of the oral cavity. Many bacteria residing in biofilms exhibit an enhanced antibiotic tolerance, which grants intrinsically susceptible microorganisms to survive lethal concentrations of antibiotics. In this study, we examined the effects of two endogenous human antimicrobial peptides, LL-37 and human Lactoferricin, on the antibiotic drug efficacy of amoxicillin, clindamycin and metronidazole in two types of polymicrobial biofilms, which aimed to represent frequent oral diseases: (1) facultative anaerobic (Streptococcus mutans, Streptococcus sanguinis, Actinomyces naeslundii) and (2) obligate anaerobic biofilms (Veillonella parvula, Parvimonas micra, Fusobacterium nucleatum). LL-37 and Lactoferricin enhanced the anti-biofilm effect of amoxicillin and clindamycin in facultative anaerobic biofilms. Metronidazole alone was ineffective against facultative anaerobic biofilms, but the presence of LL-37 and Lactoferricin led to a greater biofilm reduction. Obligate anaerobic biofilms showed an increased drug tolerance to amoxicillin and clindamycin, presumably due to metabolic downshifts of the bacteria residing within the biofilm. However, when combined with LL-37 or Lactoferricin, the reduction of obligate anaerobic biofilms was markedly enhanced for all antibiotics, even for amoxicillin and clindamycin. Furthermore, our results suggest that antimicrobial peptides enhance the dispersion of matured biofilms, which may be one of their mechanisms for targeting biofilms. In summary, our study proves that antimicrobial peptides can serve as an auxiliary treatment strategy for combatting enhanced antibiotic tolerance in bacterial biofilms.

First comparison of a new dynamic navigation system and surgical guides for implantology: an in vitro study.

AIM: The transfer of 3D implant position planning to the clinical site is challenging. The aim of this study was to compare in vitro this transfer accuracy by using dynamic real-time navigation or static surgical guides. MATERIALS AND METHODS: Deviations between planned and actual pilot drill positions were calculated (Denacam and NobelGuide; each n = 90), matching pre- and postoperative CBCT images: entry point, angle, tip (each 3D); depth, mesiodistal/bucco-oral entry points, and angles (each 1D). The influence of the maxilla and mandible, implant region (anterior/posterior), and marker position (ipsilateral/contralateral, Denacam only) was investigated (Mann-Whitney U test). RESULTS: No significant differences occurred regarding entry point (3D), mesiodistal/bucco-oral entry points (1D) or mesiodistal angle (1D) (P > 0.05). The angular and tip deviations (3D) were significantly smaller using Denacam (2.16 ± 0.59 degrees, 0.80 ± 0.55 mm; NobelGuide 2.54 ± 1.19 degrees, 1.09 ± 0.56 mm; P = 0.024, P < 0.0001). The deviations in depth and bucco-oral angle (1D) were significantly smaller using NobelGuide (1.05 ± 0.50 mm, 1.02 ± 1.16 degrees; Denacam 1.50 ± 0.64 mm, 1.51 ± 0.82 degrees; P < 0.0001). Significantly smaller deviations occurred within the mandible (Denacam, 5/8 parameters). Region and marker position showed no influence. No distinct influences were found with the use of NobelGuide. CONCLUSION: Denacam might possibly be a promising alternative to static surgical guides.

Elution behavior of a 3D-printed, milled and conventional resin-based occlusal splint material.

OBJECTIVE: The elution of unpolymerized (co-)monomers and additives from methacrylic resin-based materials like polymethyl methacrylate (PMMA) can cause adverse side effects, such as mutagenicity, teratogenicity, genotoxicity, cytotoxicity and estrogenic activity. The aim of this study was to quantify the release and the cytotoxicity of residual (co-)monomers and additives from PMMA-based splint materials under consideration of real splint sizes. Three different materials used for additive (3D printing), subtractive (milling) and conventional (powder and liquid) manufacturing were examined. METHODS: The splint materials SHERAprint-ortho plus (additive), SHERAeco-disc PM20 (subtractive) and SHERAORTHOMER (conventional) were analysed. 16 (n = 4) sample discs of each material (6 mm diameter and 2 mm height) were polished on the circular and one cross-section area and then eluted in both distilled water and methanol. The discs were incubated at 37 °C for 24 h or 72 h and subsequently analysed by gas chromatography/mass spectrometry (GC/MS) for specifying and quantifying released compounds. XTT-based cell viability assays with human gingival fibroblasts (HGFs) were performed for Tetrahydrofurfuryl methacrylate (THFMA), 1,4-Butylene glycol dimethacrylate (BDDMA) and Tripropylenglycol diacrylate (TPGDA). In order to project the disc size to actual splint sizes in a worst-case scenario, lower and upper jaw occlusal splints were designed and volumes and surfaces were measured. RESULTS: For SHERAeco-disc PM20 and for SHERAORTHOMER no elution was determined in water. SHERAprint-ortho plus eluted the highest THFMA concentration of 7.47 µmol/l ±2,77 µmol/l after 72 h in water. Six (co-)monomers and five additives were detected in the methanol eluates of all three materials tested. The XTT-based cell viability assays resulted in a EC50 of 3006 ± 408 µmol/l for THFMA, 2569.5 ± 308 µmol/l for BDDMA and 596.7 ± 88 µmol/l for TPGDA. SIGNIFICANCE: With the solvent methanol, released components from the investigated splint materials exceeded cytotoxic concentrations in HGFs calculated for a worst-case scenario in splint size. In the water eluates only the methacrylate THFMA could be determined from SHERAprint-ortho plus in concentrations below cytotoxic levels in HGFs.

Effect of polyhexanide as antiseptic mouth rinse against oral pathogens in an in vitro biofilm model.

OBJECTIVE: The aim of this study was to evaluate the anti-biofilm effect of polyhexanide mouth rinses against oral pathogens in vitro. MATERIAL AND METHODS: Biofilms of Candida albicans, Streptococcus mutans, Actinomyces naeslundii, Aggregatibacter actinomycetemcomitans, methicillin-resistant Staphylococcus aureus and Fusobacterium nucleatum were grown on 10 mm diameter hydroxyapatite discs for 5 days. Biofilms were exposed to test substances for 30 s (ProntOral, polyhexanide 0.15%, chlorhexidine 0.2%). Another test set simulating blood contamination in the oral cavity was performed by submerging the discs in defibrinated sheep blood prior to antimicrobial exposure. Biofilm mass was determined via crystal violet staining. The proliferation potency of the cells after antimicrobial exposure was evaluated by plating serially diluted suspensions from extracted biofilms on agar plates and determining the number of colony-forming units (CFU/ml). Mann-Whitney-U, Kruskal-Wallis and Dunn's test were used for statistical analyses. RESULTS: Regardless of blood contamination ProntOral led to a significant reduction of biofilm mass in all strains. Chlorhexidine and polyhexanide reduced biofilm mass in five out of six strains and in only four strains after blood contamination. All agents significantly reduced CFU/ml from S. mutans, A. actinomycetemcomitans and F. nucleatum biofilms. C. albicans and S. aureus biofilms were only affected by ProntOral and polyhexanide. None of the antiseptics significantly reduced the CFU/ml for A. naeslundii biofilms. After blood contamination ProntOral and polyhexanide significantly reduced CFU/ml in all strains, whereas CHX tended to increase the CFU/ml. CONCLUSIONS: Polyhexanide mouth rinses seem to be suitable disinfectants against oral pathogens without their anti-biofilm potential being impaired by blood.

Inhibitory effect of LL-37 and human lactoferricin on growth and biofilm formation of anaerobes associated with oral diseases.

This study was conducted to evaluate the antimicrobial potential of the antimicrobial peptides (AMP) LL-37 and human Lactoferricin (LfcinH) on the planktonic growth and biofilm formation of oral pathogenic anaerobes related to caries and periodontitis. Multi-species bacterial suspensions of either facultative anaerobic bacteria (FAB: Streptococcus mutans, Streptococcus sanguinis, Actinomyces naeslundii) or obligate anaerobic bacteria (OAB: Veillonella parvula, Parvimonas micra, Fusobacterium nucleatum) were incubated with different concentrations of AMP solutions for 8 h. Planktonic growth was registered with an ATP-based cell viability assay for FAB and via plate counting for OAB. Biofilms were grown on ZrO2 discs for 4 days in a mixture of the multi-species bacterial suspensions and AMP solutions. Biofilm mass was quantified using a microtiter plate biofilm assay with crystal violet staining. An overall planktonic growth inhibition and biofilm mass reduction of FAB and OAB was registered for LL-37 and LfcinH. Significant inhibitory threshold concentrations of LL-37 were observed in all experiments (p < 0.0001). No significant threshold was observed for LfcinH. Biofilm mass of OAB was barely reduced by LfcinH. The complete mechanisms of the AMPs are not fully understood yet. While LL-37 shows promising features as potential therapeutic for biofilm-associated oral diseases, LfcinH seems unsuitable for this particular indication. For clinical AMP use, further investigations will be necessary.

Near-infrared transillumination with high dynamic range imaging for occlusal caries detection in vitro.

The aim of this study was to assess near-infrared transillumination with high dynamic range imaging (NIRT-HDRI) for occlusal caries detection in vitro and to compare it with visual inspection using the International Caries Detection and Assessment System II (ICDAS) and digital bitewing radiography (BWR). Sixty-one extracted permanent molars with sound or occlusal carious surfaces without severe cavitation were visually assembled. Two examiners assessed twice these surfaces independently using ICDAS, BWR and NIRT-HDRI. The latter was performed with a prototype consisting of two laser sources (780 nm), a CCD sensor and subsequent processing with image analysis software. Thresholds for carious surfaces, enamel and dentin lesions were defined for all methods. Micro-computed tomography served as the reference standard. Linear weighted Kappa analysis of the methods versus the reference at the threshold carious surface, enamel and dentin lesion revealed 0.59/0.08/0.12 for ICDAS, 0.37/- 0.06/0.58 for BWR and 0.33/- 0.01/0.51 for NIRT-HDRI. Sensitivity values at the three thresholds were 0.85/0.78/0.13 for ICDAS, 0.59/0.00/0.69 for BWR and 0.98/0.33/0.78 for NIRT. Specificity values at the three thresholds were 0.70/0.40/1.00 for ICDAS, 0.9./0.96/0.90 for BWR and 0.30/0.65/0.72 for NIRT-HDRI. Reliability analysis revealed substantial agreement for BWR and NIRT and almost perfect agreement for ICDAS. NIRT exhibited a strong ability to identify occlusal dental decay in general; however, it revealed a tendency towards overestimation. Its strength was the detection of dentin caries lesions compared with ICDAS and BWR. NIRT-HDRI seems to be a suitable method to detect hidden dentin caries as a supplement to visual examination.

Fracture load and chewing simulation of zirconia and stainless-steel crowns for primary molars.

Aesthetic alternatives to stainless-steel crowns for restoring primary molars attain growing interest. We studied the mechanical properties of prefabricated zirconia crowns and conventional crowns. Three brands of prefabricated zirconia crowns were compared with computer-aided design/computer-aided manufacturing (CAD/CAM) zirconia crowns, preveneered stainless-steel crowns, and conventional stainless-steel crowns regarding: (i) fracture load under each of three conditions [no pretreatment, artificial aging in saliva for 12 wk, and after chewing simulation/thermocycling (1.68 × 106 cycles/5-55°C)]; and (ii) survival rate during chewing simulation, considering decementation, fracture, chipping, fatigue cracks, and occlusal holes. Without pretreatment, the prefabricated zirconia crowns showed mean fracture load values between 893 N and 1,582 N, while the corresponding values for CAD/CAM zirconia crowns and preveneered stainless-steel crowns were 2,444 N and 6,251 N. Preveneered stainless-steel crowns showed significantly lower fracture loads after artificial aging (5,348 N after saliva aging; 3,778 N after chewing simulation) than without artificial aging, whereas the fracture load of zirconia crowns was not influenced negatively. The survival rate of the different groups of zirconia crowns and preveneered stainless-steel crowns during chewing simulation was 100%, but only 41.7% for the stainless-steel crowns. These in-vitro data suggest that prefabricated zirconia crowns are aesthetically and durable alternatives to stainless-steel crowns for primary molars.

Sensitivity of caries pathogens to antimicrobial peptides related to caries risk.

OBJECTIVES: Antimicrobial peptides (AMPs) represent important facets of the immune system controlling infectious diseases. However, pathogens show varying susceptibilities to AMPs. This study investigates the susceptibilities of strains of Streptococcus mutans (SM), Actinomyces naeslundii (AN), and Lactobacillus spp. (LB) towards AMPs and if there are correlations between the appearance of such high-risk strains and clinical caries status. MATERIAL AND METHODS: Plaque samples were collected from patients along with clinical examinations. Bacterial strains were identified via selective media, matrix-assisted laser desorption/ionization analysis-time of flight (MALDI-TOF), and arbitrary-primed-PCR (AP-PCR). Each strain was tested for susceptibility to LL-37, HBD-2, HNP-1, and HNP-3 or phosphate-buffered saline as negative control in a biofilm model on hydroxylapatite discs. Survival rates and resulting risk classification for each strain were determined. Correlations were calculated between the number of high-risk strains (all/S. mutans) appearing in patients and their clinical caries status. RESULTS: Forty-seven patients were included with mean DMFT values of 11.4 ± 8.7. A total of 8 different SM, 30 LB, and 47 AN strains were detected. One-way ANOVA indicated that type/concentration of AMPs had major influence on reductions of Lactobacilli and Actinomyces. Seventeen strains of AN, 2 of SM, and 6 of LB had low susceptibilities to AMPs. The number of such strains in patients showed significant positive correlations to the DMFT values (all p = 0.001; r = 0.452; S. mutans p < 0.0001, r = 0.558). CONCLUSION: The occurrence of low susceptible strains to AMPs seems to correlate with the individual caries status. CLINICAL RELEVANCE: The results may lead to new ways to identify individuals with increased caries risk.