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.

Two-body wear of novel monolithic lithium-silicate ceramic materials and their corresponding different antagonists.

OBJECTIVES: Evaluation of the two-body wear of lithium-silicate ceramics against different antagonists compared to a direct resin composite and human teeth. METHODS: Initial LiSi Block [LISI], IPS e.max CAD [EMA], and CEREC Tessera [TESE] were investigated and compared with direct resin composite [FILL] and human teeth [tooth]. As antagonists were used: steatite, ceramic, and human enamel. The control group tooth was only tested with enamel antagonist. The combinations underwent thermomechanical aging using a chewing simulator. Material losses were calculated using GOM-analysis software. Kolmogorov-Smirnov test, Kruskal-Wallis H, Mann-Whitney-U-test with Bonferroni correction and Spearman-rho correlation were calculated. A fractographic analysis was performed. RESULTS: Within TESE, enamel antagonists led to lower restoration losses than steatite and ceramic antagonists. Within FILL, enamel and steatite antagonists caused lower material losses compared to ceramic antagonists. Against steatite antagonists, LISI showed lowest material losses. Against ceramic antagonists, the use of LISI led to lower material losses compared to FILL. Against tooth antagonists, TESE showed lower material losses than tooth and FILL and LISI lower than FILL. Within LISI, steatite antagonists showed lower material losses on the antagonist than ceramic. Within EMA, steatite antagonists showed higher material losses than ceramic ones. Within ceramic antagonists, LISI restoration material showed lower material losses than FILL and EMA. CONCLUSIONS: Regardless of the antagonist material, the material losses of LISI and EMA were comparable. However, the abrasion resistance of LISI tended to be higher than EMA. CLINICAL SIGNIFICANCE: LISI is a fully crystallized lithium-silicate ceramic and no longer needs to be processed after milling. In addition, the abrasion resistance is very good, regardless of the antagonist material chosen.

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.

Biopsies of osseous jaw lesions using 3D-printed surgical guides: a clinical study.

BACKGROUND: Bone biopsies are often necessary to make a diagnosis in the case of irregular bone structures of the jaw. A 3D-printed surgical guide may be a helpful tool for enhancing the accuracy of the biopsy and for ensuring that the tissue of interest is precisely removed for examination. This study was conducted to compare the accuracy of biopsies performed with 3D-printed surgical guides to that of free-handed biopsies. METHODS: Computed tomography scans were performed on patients with bony lesions of the lower jaw. Surgical guides were planned via computer-aided design and manufactured by a 3D-printer. Biopsies were performed with the surgical guides. Bone models of the lower jaw with geometries identical to the patients' lower jaws were produced using a 3D-printer. The jaw models were fitted into a phantom head model and free-handed biopsies were taken as controls. The accuracy of the biopsies was evaluated by comparing the parameters for the axis, angle and depth of the biopsies to the planned parameters. RESULTS: Eight patients were included. The mean deviation between the biopsy axes was significantly lower in guided procedures than in free-handed biopsies (1.4 mm ± 0.9 mm; 3.6 mm ± 1.0 mm; p = 0.0005). The mean biopsy angle deviation was also significantly lower in guided biopsies than in free-handed biopsies (6.8° ± 4.0; 15.4° ± 3.6; p = 0.0005). The biopsy depth showed no significant difference between the guided and the free-handed biopsies. CONCLUSIONS: Computer-guided biopsies allow significantly higher accuracy than free-handed procedures.

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.

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.

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.