Application of the virtual-fit method for fixed complete denture cases designed on intraoral scans: Effect of cement spacing.

OBJECTIVES: To validate the virtual-fit alignment, analyze the impact of cement spacing on internal/marginal gaps, and correlate results with conventional trueness measures. METHODS: Four dental abutment models were scanned using an industrial reference scanner (one time each), Emerald S (three times each), and Medit i700 (three times each) intraoral scanners (IOS). On each IOS scan (n = 24), three complete-arch fixed frameworks were designed with 70 or 140 µm cement space with no marginal space (groups 70 and 140) and 70 µm with an additional 20 µm space, including the margin (group 70+20). Two types of alignment were performed by GOM Inspect software. The reference and IOS scans were aligned through a conventional iterative closest point algorithm (ICP) where the penetration of the two scans was permitted into each other (conventional trueness method). Second, the computer-aided designs were superimposed with the reference scan also using an ICP, but preventing the design from virtual penetration into the model (virtual-fit method). The virtual-fit algorithm was validated by non-penetration alignment of the designs with the IOS scans. Internal and marginal gap was measured between the design and the abutments. The difference between spacing groups was compared by Friedman's test. A statistical correlation (Spearman's Rho Test) was computed between the measured gaps and the conventional trueness method. A significant difference was accepted at p<0.05 after the Bonferroni correction. RESULTS: The gaps deviated from the set cement space by 3-13 µm on IOS scans (validation of virtual-fit algorithm). The internal gap of the design on the reference scan was not affected by cement spacing (Emerald S, p = 0.779; Medit i700, p = 0.205). The marginal gap in groups 70 and 70+20 was significantly lower than in group 140 in Emerald S (p<0.05). In Medit i700, it was lower in the 70+20 group than in the group 70 (p<0.01) and in the group 140 (p<0.05). Some Medit i700 scans exhibited high marginal gaps within group 70 but not in groups 70 and 140. The measured gaps correlated significantly (r = 0.51-0.81, p<0.05-0.001) with the conventional trueness but were 2.6-4.6 times higher (p<0.001). CONCLUSION: Virtual-fit alignment can simulate restoration seating. A 20 µm marginal and 90 µm internal spacing could compensate for scan errors up to several hundred micrometers. However, 140 µm internal spacing is counterproductive. The conventional trueness method could only partially predict framework misfit. CLINICAL SIGNIFICANCE: The virtual-fit method can provide clinically interpretable data for intraoral scanners. Emerald S and Medit i700 intraoral scanners are suitable for fabricating complete-arch fixed tooth-supported prostheses. In addition, a slight elevation of spacing at the margin could compensate for moderate inaccuracies in a scan.

In vitro comparison of five desktop scanners and an industrial scanner in the evaluation of an intraoral scanner accuracy.

OBJECTIVES: The study aimed to compare the precision of ATOS industrial, 3ShapeE4, MeditT710, CeramillMap400, CSNeo, PlanScanLab desktop, and Mediti700 intraoral scanners. The second aim was to compare the trueness of Mediti700 assessed by ATOS and desktop scanners. METHODS: Four plastic dentate models with 7-12 abutments prepared for complete arch fixed dentures were scanned by all scanners three times. Scans were segmented to retain only the abutments. The precision and trueness were calculated by superimposing scans with the best-fit algorithm. The mean absolute distance was calculated between the scan surfaces. The precision was calculated based on the 12 repeats. Trueness was evaluated by superimposing the desktop and IOS scans to the industrial scans. IOS was also aligned with the two most accurate desktop scanners. RESULTS: The precision of 3ShapeE4 and MeditT710 (3-4µm) was only slightly lower than that of ATOS (1.7µm, p<0.001) and significantly higher than CeramillMap400, CSNeo, and PlanScanLab (6-10 µm, p<0.001). The trueness was the highest for the 3Shape E4 (12-13 µm) and Medit T710 (13-16 µm) without significant difference. They were significantly better than CeramillMap400, CSNeo, and PlanScanLab (22-31µm, p<0.001). Accordingly, the Mediti700 trueness was evaluated by ATOS, 3ShapeE4, and MeditT710. The three trueness was not significantly different; ATOS (23-26 µm), 3Shape E4 (22-25 µm), and Medit T710 (20-23 µm). CONCLUSIONS: All desktop scanners had the acceptable accuracy required for a complete arch-fixed prosthesis. The 3Shape E4 and the Medit T710 might be used as reference scanners for studying IOS accuracy. CLINICAL SIGNIFICANCE: 3ShapeE4, MeditT710, CeramillMap400, CSNeo, PlanScanLab laboratory, and Mediti700 intraoral scanners can be used for the prosthetic workflow in a complete arch. 3ShapeE4 and the MeditT710 could be used to test the accuracy of various phases of a laboratory workflow, replacing the industrial scanners.

Antibacterial properties of copper iodide-doped glass ionomer-based materials and effect of copper iodide nanoparticles on collagen degradation.

OBJECTIVES: This study investigated the antibacterial properties and micro-hardness of polyacrylic acid (PAA)-coated copper iodide (CuI) nanoparticles incorporated into glass ionomer-based materials, and the effect of PAA-CuI on collagen degradation. MATERIALS AND METHODS: PAA-CuI nanoparticles were incorporated into glass ionomer (GI), Ionofil Molar AC, and resin-modified glass ionomer (RMGI), Vitrebond, at 0.263 wt%. The antibacterial properties against Streptococcus mutans (n = 6/group) and surface micro-hardness (n = 5/group) were evaluated. Twenty dentin beams were completely demineralized in 10 wt% phosphoric acid and equally divided in two groups (n = 10/group) for incubation in simulated body fluid (SBF) or SBF containing 1 mg/ml PAA-CuI. The amount of dry mass loss and hydroxyproline (HYP) released were quantified. Kruskal-Wallis, Student's t test, two-way ANOVA, and Mann-Whitney were used to analyze the antibacterial, micro-hardness, dry mass, and HYP release data, respectively (p < 0.05). RESULTS: Addition of PAA-CuI nanoparticles into the glass ionomer matrix yielded significant reduction (99.999 %) in the concentration of bacteria relative to the control groups. While micro-hardness values of PAA-CuI-doped GI were no different from its control, PAA-CuI-doped RMGI demonstrated significantly higher values than its control. A significant decrease in dry mass weight was shown only for the control beams (10.53 %, p = 0.04). Significantly less HYP was released from beams incubated in PAA-CuI relative to the control beams (p < 0.001). CONCLUSIONS: PAA-CuI nanoparticles are an effective additive to glass ionomer-based materials as they greatly enhance their antibacterial properties and reduce collagen degradation without an adverse effect on their mechanical properties. CLINICAL RELEVANCE: The use of copper-doped glass ionomer-based materials under composite restorations may contribute to an increased longevity of adhesive restorations, because of their enhanced antibacterial properties and reduced collagen degradation.

Lipoteichoic acid (LTA) and lipopolysaccharides (LPS) from periodontal pathogenic bacteria facilitate oncogenic herpesvirus infection within primary oral cells.

Kaposi's sarcoma (KS) remains the most common tumor arising in patients with HIV/AIDS, and involvement of the oral cavity represents one of the most common clinical manifestations of this tumor. HIV infection incurs an increased risk for periodontal diseases and oral carriage of a variety of bacteria. Whether interactions involving pathogenic bacteria and oncogenic viruses in the local environment facilitate replication or maintenance of these viruses in the oral cavity remains unknown. In the current study, our data indicate that pretreatment of primary human oral fibroblasts with two prototypical pathogen-associated molecular patterns (PAMPs) produced by oral pathogenic bacteria-lipoteichoic acid (LTA) and lipopolysaccharide (LPS), increase KSHV entry and subsequent viral latent gene expression during de novo infection. Further experiments demonstrate that the underlying mechanisms induced by LTA and/or LPS include upregulation of cellular receptor, increasing production of reactive oxygen species (ROS), and activating intracellular signaling pathways such as MAPK and NF-κB, and all of which are closely associated with KSHV entry or gene expression within oral cells. Based on these findings, we hope to provide the framework of developing novel targeted approaches for treatment and prevention of oral KSHV infection and KS development in high-risk HIV-positive patients.

Kaposi's sarcoma-associated herpesvirus suppression of DUSP1 facilitates cellular pathogenesis following de novo infection.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma (KS), and KSHV activation of mitogen-activated protein kinases (MAPKs) initiates a number of key pathogenic determinants of KS. Direct inhibition of signal transduction as a therapeutic approach presents several challenges, and a better understanding of KSHV-induced mechanisms regulating MAPK activation may facilitate the development of new treatment or prevention strategies for KS. MAPK phosphatases, including dual-specificity phosphatase-1 (DUSP1), negatively regulate signal transduction and cytokine activation through MAPK dephosphorylation or interference with effector molecule binding to MAPKs, including the extracellular signal-regulated kinase (ERK). We found that ERK-dependent latent viral gene expression, the induction of promigratory factors, and cell invasiveness following de novo infection of primary human endothelial cells are in part dependent on KSHV suppression of DUSP1 expression during de novo infection. KSHV-encoded miR-K12-11 upregulates the expression of xCT (an amino acid transporter and KSHV fusion/entry receptor), and existing data indicate a role for xCT in the regulation of 14-3-3ß, a transcriptional repressor of DUSP1. We found that miR-K12-11 induces endothelial cell secretion of promigratory factors and cell invasiveness through upregulation of xCT-dependent, 14-3-3ß-mediated suppression of DUSP1. Finally, proof-of-principle experiments revealed that pharmacologic upregulation of DUSP1 inhibits the induction of promigratory factors and cell invasiveness during de novo KSHV infection. These data reveal an indirect role for miR-K12-11 in the regulation of DUSP1 and downstream pathogenesis.

Predicting marginal fit of CAD/CAM crowns based on the presence or absence of common preparation errors.

STATEMENT OF PROBLEM: Confusion exists as to what constitutes an ideal ceramic crown preparation and whether certain deviations from the ideal can affect the marginal fit of the milled restoration. PURPOSE: This study evaluated the marginal gap of E4D crowns fabricated on preparations completed by clinicians with varying levels of expertise to identify whether common errors affect marginal fit. MATERIAL AND METHODS: The fit of 75 crowns fabricated with the E4D system on preparations of varying quality were examined for marginal fit by using the replica technique. These same preparations were then visually examined for common criteria for ceramic restorations and placed in one of 3 categories: excellent, fair, or poor. These visual examinations sought the presence of common preparation errors, particularly those involving the finish line. The average marginal gap values and standard deviations were calculated for each category, and the Kruskal-Wallis test was used to determine significance. RESULTS: The results showed a statistically significant correlation between the marginal fit of a CAD/CAM fabricated crown and the quality of the preparation. The mean marginal gap of the crowns fabricated on ideal preparations was 38.5 µm, those considered fair had a mean marginal gap of 58.3 µm, while those categorized as poor averaged 90.1 µm. The fit differences among all 3 groups were statistically significant (P<.05). CONCLUSIONS: Within the limitations of this in vitro study, it can be concluded that preparation quality has a significant impact on marginal gap on crowns fabricated with a CAD/CAM system.

Kaposi sarcoma-associated herpesvirus (KSHV) induces a functional tumor-associated phenotype for oral fibroblasts.

The Kaposi sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi sarcoma (KS), the most common HIV/AIDS-associated tumor worldwide. Involvement of the oral cavity portends a poor prognosis for patients with KS, but mechanisms for KSHV regulation of the oral tumor microenvironment are largely unknown. Infiltrating fibroblasts are found with KS lesions, and KSHV establishes latent infection within human primary fibroblasts in vitro, but contributions for KSHV-infected fibroblasts to the KS microenvironment have not been previously characterized. Secretion of pro-migratory factors and intratumoral invasion are characteristics of tumor-associated fibroblasts (TAF) found in the microenvironment of non-viral malignancies. In the present study, we show that latent KSHV infection of primary human fibroblasts isolated from the oral cavity enhances their secretion of KS-promoting cytokines and intrinsic invasiveness through VEGF-dependent mechanisms. Moreover, we find that KSHV induces these effects through Sp1- and Egr2-dependent transcriptional activation of the Extracellular Matrix MetalloPRoteinase INducer (emmprin). These data implicate KSHV activation of emmprin in the induction of a "TAF-like" phenotype for oral fibroblasts in the KS microenvironment and support the potential utility of targeting TAFs and/or emmprin in the treatment of oral KS.

Interactions between Hsp90 and oncogenic viruses: implications for viral cancer therapeutics.

Oncogenic viruses are the etiologic agents for a significant proportion of human cancers, but effective therapies and preventative strategies are lacking for the majority of virus-associated cancers. Targeting of virus-induced signal transduction or virus-host protein interactions may offer novel therapeutic strategies for viral cancers. Heat shock protein 90 (Hsp90) is a well-characterized, multifunctional molecular chaperone involved in regulation of signal transduction, transcriptional activation, oncogenic protein stabilization, and neovascularization-pathogenic elements relevant to viral cancer pathogenesis. This review will summarize mechanistic concepts involving regulation of viral oncogenesis by both intracellular and extracellular Hsp90, as well as current therapeutic implications of these data.

Extracellular Hsp90 serves as a co-factor for NF-κB activation and cellular pathogenesis induced by an oncogenic herpesvirus.

The Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma (KS)-the most common tumor associated with HIV infection and an important cause of morbidity and mortality in this patient population. The majority of patients with KS exhibit little or no clinical response to existing therapies. The nuclear factor-kappaB (NF-κB) family of transcription factors plays a critical role in facilitating cancer pathogenesis associated with oncogenic viruses, and a better understanding of how cellular factors regulate NF-κB activation in the context of KSHV infection may facilitate development of new therapies for KS. Existing data implicate heat shock protein-90 associated with the cell surface (csHsp90) as a co-factor in cancer cell migration and invasion, and we recently reported that csHsp90 serves as a co-factor for mitogen-activated protein kinase (MAPK) activation during de novo KSHV infection. However, whether csHsp90 regulates NF-κB activation, or cellular pathogenesis associated with KS, has not been established. We have found that csHsp90 serves as an important co-factor for canonical NF-κB activation by KSHV during de novo infection of primary human cells relevant to KS. Furthermore, our correlative functional studies reveal that csHsp90 inhibition suppresses KSHV-induced, NF-κB-dependent secretion of the pro-migratory factors interleukin-8 and vascular endothelial growth factor as well as invasiveness for primary cells following de novo infection. These data implicate csHsp90 in KSHV-mediated activation of NF-κB and associated pathogenesis, and support the potential utility of targeting csHsp90 as a therapeutic approach for KS.

Extracellular Hsp90 serves as a co-factor for MAPK activation and latent viral gene expression during de novo infection by KSHV.

The Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma (KS), an important cause of morbidity and mortality in immunocompromised patients. KSHV interaction with the cell membrane triggers activation of specific intracellular signal transduction pathways to facilitate virus entry, nuclear trafficking, and ultimately viral oncogene expression. Extracellular heat shock protein 90 localizes to the cell surface (csHsp90) and facilitates signal transduction in cancer cell lines, but whether csHsp90 assists in the coordination of KSHV gene expression through these or other mechanisms is unknown. Using a recently characterized non-permeable inhibitor specifically targeting csHsp90 and Hsp90-specific antibodies, we show that csHsp90 inhibition suppresses KSHV gene expression during de novo infection, and that this effect is mediated largely through the inhibition of mitogen-activated protein kinase (MAPK) activation by KSHV. Moreover, we show that targeting csHsp90 reduces constitutive MAPK expression and the release of infectious viral particles by patient-derived, KSHV-infected primary effusion lymphoma cells. These data suggest that csHsp90 serves as an important co-factor for KSHV-initiated MAPK activation and provide proof-of-concept for the potential benefit of targeting csHsp90 for the treatment or prevention of KSHV-associated illnesses.