Fully digital workflow of an occlusal device including digital facebow record: A clinical report.

In many current digital workflows for the production of occlusal devices and dentures, the traditional facebow record for the individual determination of the hinge axis has been omitted. A novel digital facebow record procedure including a scannable occlusal fork is described as a straightforward to use, cost-effective, and less time-consuming alternative to close this gap in fully digital prosthetic workflows.

Human Gingival Fibroblast Adhesion and Proliferation on Hydroxyapatite-Coated Zirconia Abutment Surfaces.

Applying antibacterial coatings to dental implant materials seems reasonable but can have negative influences on desired cell adhesion and healing. In this study, zirconia abutment specimens interacting with gingival tissue were used. The aim was to compare the influence of machined or coated zirconia surfaces on the adhesion and proliferation of human gingival fibroblasts (HGF-1). Surface modifications were performed using atmospheric plasma coating with hydroxyapatite, zinc, and copper. Zirconia specimens were divided into four groups: hydroxyapatite, hydroxyapatite with zinc oxide (ZnO), hydroxyapatite with copper (Cu), and an untreated machined surface. After the characterization of the surface conditions, the morphology of adhered HGF-1 was determined by fluorescence staining and subjected to statistical evaluation. The visual analysis of cell morphology by SEM showed flat, polygonal, and largely adherent fibroblast cells in the untreated group, while round to partially flat cells were recorded in the groups with hydroxyapatite, hydroxyapatite + ZnO, and hydroxyapatite + Cu. The cell membranes in the hydroxyapatite + ZnO and hydroxyapatite + Cu groups appeared porous. The results show that HGF-1 adhere and proliferate well on machined zirconia, while plasma coating with hydroxyapatite or hydroxyapatite mixtures does not lead to increased adhesion or proliferation.

Enhanced Antiviral Function of Magnesium Chloride-Modified Heparin on a Broad Spectrum of Viruses.

Previous studies reported on the broad-spectrum antiviral function of heparin. Here we investigated the antiviral function of magnesium-modified heparin and found that modified heparin displayed a significantly enhanced antiviral function against human adenovirus (HAdV) in immortalized and primary cells. Nuclear magnetic resonance analyses revealed a conformational change of heparin when complexed with magnesium. To broadly explore this discovery, we tested the antiviral function of modified heparin against herpes simplex virus type 1 (HSV-1) and found that the replication of HSV-1 was even further decreased compared to aciclovir. Moreover, we investigated the antiviral effect against the new severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) and measured a 55-fold decreased viral load in the supernatant of infected cells associated with a 38-fold decrease in virus growth. The advantage of our modified heparin is an increased antiviral effect compared to regular heparin.

Cold Atmospheric Plasma Improves Shear Bond Strength of Veneering Composite to Zirconia.

Chipping of veneering is the most common clinical complication for zirconia restorations. Veneering composite could be a promising alternative to renew restorations. Zirconia discs (3-YSZ) were prepared with varying surface treatments and bonded to indirect composite as follows: air abrasion and Scotchbond Universal (A/SU); air abrasion and Clearfil Ceramic Primer (A/C); air abrasion and MKZ Primer (A/M); air abrasion and Monobond Plus (A/MP); silica-coating and Scotchbond Universal (S/SU); air abrasion (AP/SU), additional cold atmospheric plasma treatment, and Scotchbond Universal. An indirect composite material was then applied to the zirconia specimens. Specimens were divided into subgroups for short-term (14 days storage at 37 °C and 5000 thermal cycles) and long-term (250 days storage and 37,500 thermal cycles) artificial aging. Shear bond strength measurement (SBS) was performed, and data were analyzed by Kruskal-Wallis-test and multiple comparison testing with Dunn's correction (p ≤ 0.05). The median SBS values (MPa) of short- and long-term artificial aging were: 3.09/1.36 (A/SU); 0.77/1.43 (S/SU); 2.82/2.15 (AP/SU); 1.97/1.80 (A/C); 2.01/1.58 (A/M); and 1.70/1.68 (A/MP). For short-term artificial aging A/SU showed the highest median SBS values, whereas in the long-term trial, AP/SU showed the highest values and the difference was significant. A prolonged artificial aging decreased SBS in all groups, except S/SU. In summary, treatment with CAP can improve SBS in the long-term.

Shear bond strength of veneering composite to high performance polymers.

High performance polymers like PEEK (polyetheretherketone) and FRC (fiberreinforced composite) could substitute metallic alloys for removable partial dentures. However, these polymers require aesthetic veneering. This study was to determine the bond strength to direct composite. Specimens made of PEEK and FRC were produced and air-abraded (50 µm aluminum-oxide). Specimens were allocated to four experimental groups: Luxatemp Glaze & Bond, Scotchbond Universal, SR Nexco Connect and iBond Universal. Specimens were divided into three subgroups for short-term, long-term and no artificial aging and shear bond strength (SBS) was evaluated. SBS of specimens made of PEEK with no artificial aging showed values between 10.79-14.00 MPa, short-term artificial aging resulted in values between 3.78-13.85 MPa and after long-term artificial aging SBS decreased to 0-8.75 MPa. SBS measurement of FRC specimens resulted in values between 9.83-12.1 MPa without aging, after short-term artificial aging values decreased to 8.36-11.98 MPa and after long-term aging SBS showed a degradation to 4.52-7.82 MPa.

High-Throughput Cloning and Characterization of Emerging Adenovirus Types 70, 73, 74, and 75.

Recently an increasing number of new adenovirus types associated with type-dependent pathogenicity have been identified. However, identification of these clinical isolates represents the very first step to characterize novel pathogens. For deeper analyses, these adenoviruses need to be further characterized in basic virology experiments or they could be applied in translational research. To achieve this goal, it is essential to get genetic access and to enable genetic modification of these novel adenovirus genomes (deletion, insertion, and mutation). Here we demonstrate a high-throughput approach to get genetic access to new adenoviruses via homologous recombination. We first defined the cloning conditions regarding homology arm-length and input adenoviral genome amounts. Then we cloned four naturally occurring adenoviruses (Ad70, Ad73, Ad74, and Ad75) into easy-to-manipulate plasmids and genetically modified them by reporter gene insertion. Three recombinant adenoviruses (Ad70, Ad73, and Ad74) containing a reporter cassette were successfully reconstituted. These novel reporter-labeled adenoviruses were further characterized using the inserted luciferase reporter with respect to receptor usage, presence of anti-adenovirus antibodies, and tropism in vitro. The identified receptor usage, the relatively low prevalence of anti-adenovirus antibodies, and the various cancer cell line transduction pattern are important features of these new pathogens providing essential information for their therapeutic application.

Transient Chimeric Ad5/37 Fiber Enhances NK-92 Carrier Cell-Mediated Delivery of Oncolytic Adenovirus Type 5 to Tumor Cells.

Methods for customizing and improving virus vector tropism are limited. In this study, we introduce a microRNA (miRNA)-regulated molecular method to enhance vector transduction without genome alteration. Based on the importance of adenovirus (Ad) vectors for cancer and gene treatment, we exemplified this technology for an Ad type 5 (Ad5) vector temporally carrying a knob from Ad37. We constructed a producer cell line stably expressing a fused Ad5/37 chimeric fiber comprising the Ad5 shaft-tail and the Ad37 knob and a miRNA inhibiting Ad5 knob expression (HEK293-Ad5/37-miRNA). The chimeric Ad5/37 vector resulted in enhanced transduction rates in Ad37 adequately and Ad5 poorly transduced cells. Particularly, encapsidation of the oncolytic Ad5-human telomerase reverse transcriptase (hTERT) vector genome into the chimeric Ad5/37 capsid showed efficient transduction of NK-92 carrier cells. These infected carrier cells then delivered the oncolytic vector to tumor cells, which resulted in enhanced Ad5-hTERT-mediated tumor cell killing. We show that this transiently capsid-modified chimeric vector carrying an Ad5 genome displayed higher transduction efficiencies of natural killer cell-derived NK-92 cells utilized as carriers in cancer immune therapy. In summary, transiently modified adenoviral vectors will have important implications for cancer and gene therapy.

Identification of novel human adenovirus candidates using the coxsackievirus and adenovirus receptor for cell entry.

BACKGROUND: There are over 100 known human adenovirus (HAdV) types, which are able to cause a broad variety of different self-limiting but also lethal diseases especially in immunocompromised patients. Only limited information about the pathogenesis and biology of the majority of these virus types is available. In the present study, we performed a systematic screen for coxsackievirus and adenovirus receptor (CAR)-usage of a large spectrum of HAdV types. METHODS: To study receptor usage we utilized a recombinant HAdV library containing HAdV genomes tagged with a luciferase and GFP encoding transgene. We infected CHO-CAR cells stably expressing the CAR receptor and to much information with tagged viruses (HAdV3, 14, 16, 50, 10, 24, 27, 37 and 69) and measured luciferase expression levels 26 and for some viruses (AdV10, - 24 and - 27) 52 h post-infection. As positive control, we applied human adenovirus type 5 (HAdV5) known to use the CAR receptor for cell entry. For viruses replication studies on genome level we applied digital PCR. RESULTS: Infection of CHO-CAR and CHO-K1 cells at various virus particle numbers per cell (vpc) revealed that HAdV10, 24, and 27 showed similar or decreased luciferase expression levels in the presence of CAR. In contrast, HAdV3, 14, 16, 50, 37 and 69 resulted in increased luciferase expression levels in our initial screening experiments. CAR usage of HAdV3, 14, 50, and 69 was not studied before, and therefore we experimentally confirmed CAR usage for these HAdV as novel viruses utilizing CAR as a receptor. To rule out that replication of HAdV in transduced CHO cells is responsible for increased transduction rates we performed replication assays on virus genome level, which revealed that there is no HAdV replication. CONCLUSION: In the present study, we screened a HAdV library and identified novel human HAdV using the CAR receptor. To our knowledge, this is the first description of CAR usage for HAdV 3, 14, 50, and 69.

Cold atmospheric plasma as antiviral therapy - effect on human herpes simplex virus type 1.

In previous studies, cold atmospheric plasma (CAP) was explored as an antibacterial and antiviral agent for the treatment of chronic wounds. The aim of the present study was to investigate whether CAP may also be suitable as an antiviral therapy against herpes simplex virus type 1 (HSV-1). HSV-1 most frequently manifests as recurrent herpes labialis, but it can also cause encephalitis, conjunctivitis or herpes neonatorum as a perinatal infection. HSV-1 encoding the reporter gene GFP was propagated. The CAP dose for HSV-1 treatment was gradually increased, ranging from 0-150 s, and aciclovir was used as a positive control. After CAP treatment, the virus suspension was applied to a standard HSV research cell line (Vero cells) and the neuroblastoma cell line SH-SY5Y as a model for neuronal infection. The results showed that plasma treatment had a negligible antiviral effect on HSV-1 in both Vero- and SH-SY5Y cells at high dose. However, when we lowered the viral load 100-fold, we observed a significantly decreased number of internalized HSV-1 genomes 3 h post-infection for CAP-treated viruses. This difference was less pronounced with respect to GFP expression levels 24 h post-infection, which was in sharp contrast to the acyclovir-treated positive control, for which the viral load was reduced from 95 to 25%. In summary, we observed a low but measurable antiviral effect of CAP on HSV-1.

State-of-the-art human adenovirus vectorology for therapeutic approaches.

Human adenoviruses (Ads) have long been studied in the basic virology field and are exploited as vectors for gene therapy, vaccination, and oncolytic therapy. Ads are usually mild pathogens, but they can cause severe infections and symptoms in immunocompromised individuals. Ads show a large natural diversity and a broad spectrum of hosts. However, replication-competent and replication-deficient Ad vectors with therapeutic applications have been built mainly starting from human Ad type 5, because generating vectors from other human and animal Ads has proven challenging. This review provides an updated overview of vectors that are not derived from human Ad type 5. We discuss genetic engineering techniques for getting access to the natural diversity of human Ads and for vectorization of alternative Ad types. A catalogue of currently available vectorized human Ads and translational applications thereof is also compiled. We conclude with a perspective on Ad vectorology that looks into the future of Ad vectors in translational medicine.

Effect of cold atmospheric plasma (CAP) on human adenoviruses is adenovirus type-dependent.

More than 70 human adenovirus types were identified divided into 7 different species (A-G). Diseases caused by human adenoviruses are type-dependent and can range from mild to severe respiratory infections, gastrointestinal infections or eye infections such as epidemic keratoconjunctivitis. Unfortunately there is no specific anti-adenovirus therapy available. Here we addressed the question whether treatment with cold atmospheric plasma (CAP) for anti-adenoviral therapy such as virus-mediated ulcerations may be feasible. CAP has already been explored for the treatment of dermatological diseases such as chronic wounds. To investigate whether CAP is an effective antiviral tool, purified human adenovirus types derived from different human adenovirus species (HAdV -4, -5, -20, -35, -37, -50) tagged with luciferase were treated with defined dosages of plasma. The CAP treatment was varied by incrementally increasing the time span of CAP treatment. After CAP treatment, the virus containing solution was added to eukaryotic cells and the viral load was determined by measurement of luciferase expression levels. Through the plasma treatment the adenovirus driven luciferase expression directly correlating with adenovirus transduction efficiencies could be reduced for HAdV-5 and HAdV-37. Plasma treatment had no influence on adenovirus derived luciferase expression levels for HAdV-4 and HAdV-50 and it even had a positive effect on luciferase expression levels for HAdV-20 and HAdV-35. These results suggest that CAP has a type dependent effect on adenoviruses and that infectivity can be even increased for certain adenovirus types. Further studies should address the mechanisms behind this phenomenon. In summary we demonstrate that CAP may represent an interesting option for antiviral treatment in a virus type dependent manner.

Relative Clinical Success of Bis-Acryl Composite Provisional Crowns.

The high demands on the clinical performance of a single-tooth provisional restoration necessitate that said performance be examined. The authors evaluated 24 teeth evaluated. Two test groups received bis-acrylic composite for provisional crowns (n = 12) and a single-unit, self-supporting, malleable, light-curing composite crown (n = 12). Final crowns (n = 24) served as paired controls. Evaluation of clinical success was measured using previously selected subcategories of FDI criteria. Statistical analysis was performed using McNemar's Test (α = 0.05). The null hypothesis of no improvement of the definitive crown relative to the provisionals was rejected at the 5% significance level (McNemar's P value <.001, Bonferroni corrected). Of the provisional crowns, 75% received a clinically insufficient valuation, while only 8% of the definitive crowns did. The authors concluded that, independent of the manufacturing process, a bis-acrylic composite provisional crown cannot serve as a replacement for a conventionally manufactured definitive crown.