Effect of virtual cement space and restorative materials on the adaptation of CAD-CAM endocrowns.

BACKGROUND: This study aimed to evaluate the effect of virtual cement space and restorative materials on the fit of computer-aided design and computer-aided manufacturing (CAD-CAM) endocrowns. METHODS: A mandibular first molar tooth model received a butt joint margin endocrown preparation with a 2-mm occlusal thickness. Then, using a 3D-printing system, 120 copies of this prepared die were printed and assigned equally to three groups with different cement space settings (30, 60, and 120 µm) during the chairside CAD design. In the milling process, CAD-based models with a particular space setting were subdivided into four groups (n = 10) and fabricated from different CAD-CAM materials: Vita Suprinity (VS), Celtra Duo (CD), Lava Ultimate (LU), and Grandio blocs (GR). Finally, the endocrowns were stabilized over their corresponding models with siloxane and subjected to micro-computed tomography to measure the fit. RESULTS: The cement space that was predesigned at 30 µm generated the largest marginal discrepancy (from 144.68 ± 22.43 µm to 174.36 ± 22.78 µm), which was significantly different from those at 60 µm and 120 µm (p < 0.001). The combination of VS or CD with a pre-setting cement space of 60 µm and the combination of LU or GR with a cement space of 120 µm showed better agreement between the predesigned and actual measured marginal gap widths. For internal adaptation, only the cement space set to 30 µm exceeded the clinically acceptable threshold (200 µm). CONCLUSIONS: The setting of the cement space and restorative material significantly affected the marginal adaptation of CAD-CAM endocrown restorations. Considering the discrepancy between design and reality, different virtual cement spaces should be applied to ceramic and resin composite materials.

Effect of Heat Softening versus Ultrasonic Removal of Root-End Gutta-Percha on the Quality of Root-End Preparation for Endodontic Microsurgery.

This study is aimed at comparing the efficacy of heat softening and ultrasonic removal of root-end gutta-percha during retrograde preparation for root apical microsurgery. Recently extracted single-rooted premolars (n = 40) were prepared with standardized endodontically treated and root-end resection and then randomly divided into four groups that received root-end cavity preparation using four different instruments: a diamond-coated ultrasonic tip (AS3D; SACTEON, Paris, France), AS3D with the modified plugger of cordless heat carrier (MSE; B&L Biotech, Bala Cynwyd, PA, USA), stainless steel ultrasonic tip (Jetip-2; B&L Biotech, Bala Cynwyd, PA, USA), or Jetip-2 with MSE. The time required for root-end preparation was recorded. The root apex samples were examined by scanning electron microscopy to assess root surface microcracks and marginal integrity. The remnants of gutta-percha on the cavity walls were evaluated using a stereomicroscope. Statistical analysis was performed using Student's t-test and Wilcoxon rank-sum test (P < 0.05). Usage of MSE with Jetip-2 significantly reduced preparation time from 99.8 ± 6.8 s to 32.4 ± 1.0 s (P = 0.009), the number of microcracks from 42 to 13 (P = 0.036), and the remnants of gutta-percha from 80% to 20% (P = 0.041). Similar results were observed for AS3D with MSE. The heat softening of MSE was effective in cleaning gutta-percha, thus greatly improving the efficiency of the root-end preparation, thereby producing a root-end cavity with fewer microcracks and better marginal integrity.

Case Report: A Novel COL1A1 Missense Mutation Associated With Dentineogenesis Imperfecta Type I.

Background: Osteogenesis imperfecta (OI) is a clinical and genetic disorder that results in bone fragility, blue sclerae and dentineogenesis imperfecta (DGI), which is mainly caused by a mutation in the COL1A1 or COL1A2 genes, which encode type I procollagen. Case Report: A missense mutation (c.1463G > C) in exon 22 of the COL1A1 gene was found using whole-exome sequencing. However, the cases reported herein only exhibited a clinical DGI-I phenotype. There were no cases of bone disease or any other common abnormal symptom caused by a COL1A1 mutation. In addition, the ultrastructural analysis of the tooth affected with non-syndromic DGI-I showed that the abnormal dentine was accompanied by the disruption of odontoblast polarization, a reduced number of odontoblasts, a reduction in hardness and elasticity, and the loss of dentinal tubules, suggesting a severe developmental disorder. We also investigated the odontoblast differentiation ability using dental pulp stem cells (DPSCs) that were isolated from a patient with DGI-I and cultured. Stem cells isolated from patients with DGI-I are important to elucidate their pathogenesis and underlying mechanisms to develop regenerative therapies. Conclusion: This study can provide new insights into the phenotype-genotype association in collagen-associated diseases and improve the clinical diagnosis of OI/DGI-I.

Enhanced therapeutic efficacy of Listeria-based cancer vaccine with codon-optimized HPV16 E7.

Cervical cancer is a leading cause of high mortality in women in developing countries and has a serious impact on women's health. Human papilloma virus (HPV) prophylactic vaccines have been produced and may hold promise for reducing the incidence of cervical cancer. However, the limitations of current HPV vaccine strategies make the development of HPV therapeutic vaccines particularly important for the treatment of HPV related lesions. Our previous work has demonstrated that LM4Δhly::E7 was safe and effective in inducing antitumor effect by antigen-specific cellular immune responses and direct killing of tumor cell on a cervical cancer model. In this study, the codon usage effect of a novel Listeria-based cervical cancer vaccine LM4Δhly::E7-1, was evaluated for effects of codon-optimized E7 expression, cellular immune response and therapeutic efficacy in a tumor-bearing murine model. Our data demonstrated that up-regulated expression of E7 was strikingly elevated by codon usage optimization, and thus induced significantly higher Th1-biased immunity, lymphocyte proliferation, and strong specific CTL activity ex-vivo compared with LM4Δhly::E7-treated mice. Furthermore, LM4Δhly::E7-1 enhanced a remarkable therapeutic effect in establishing tumors. Taken together, our results suggest that codon usage optimization is an important consideration in constructing live bacterial-vectored vaccines and is required for promoting effective T cell responses.

inlF Enhances Listeria monocytogenes Early-Stage Infection by Inhibiting the Inflammatory Response.

The internalin family proteins, which carry the leucine repeat region structural motif, play diverse roles in Listeria monocytogenes (Lm) infection and pathogenesis. Although Internalin F, encoded by inlF, was identified more than 20 years ago, its role in the Lm anti-inflammatory response remains unknown. Lm serotype 4b isolates are associated with the majority of listeriosis outbreaks, but the function of InlF in these strains is not fully understood. In this study, we aimed to elucidate the role of inlF in modulating the inflammatory response and pathogenesis of the 4b strain Lm NTSN. Strikingly, although inlF was highly expressed at the transcriptional level during infection of five non-phagocytic cell types, it was not involved in adherence or invasion. Conversely, inlF did contributed to Lm adhesion and invasion of macrophages, and dramatically suppressed the expression of pro-inflammatory cytokines interleukin (IL)-1ß and tumor necrosis factor (TNF-α). Consistent with the in vitro results, during Lm infection mice, inlF significantly inhibited the expression of IL-1ß and IL-6 in the spleen, as well as IL-1ß, IL-6, and TNF-α in the liver. More importantly, inlF contributed to Lm colonization in the spleen, liver, and ileum during the early stage of mouse infection via intragastric administration, inducing severe inflammatory injury and histopathologic changes in the late stage. To our knowledge, this is the first report to demonstrate that inlF mediates the inhibition of the pro-inflammatory response and contributes to the colonization and survival of Lm during the early stage of infection in mice. Our research partly explains the high pathogenicity of serovar 4b strains and will lead to new insights into the pathogenesis and immune evasion of Lm.

Biomechanical behavior of endocrown restorations with different CAD-CAM materials: A 3D finite element and in vitro analysis.

STATEMENT OF PROBLEM: The performance of endocrowns fabricated with different types of computer-aided design and computer-aided manufacturing (CAD-CAM) materials is unclear. PURPOSE: The purpose of this finite element analysis (FEA) and in vitro study was to compare and evaluate the stress distribution, failure probability, and fracture resistance of endodontically treated teeth restored with endocrowns from CAD-CAM milling blocks including ceramic, polymer-infiltrated ceramic (PICN), and composite resin. MATERIAL AND METHODS: An endodontically treated first mandibular molar restored with an endocrown was modeled by using a CAD software program and imported into an FEA software program. The model was duplicated and received restorations made from CAD-CAM blocks: Vita Suprinity (VS), IPS e.max CAD (EMX), Vita Enamic (VE), Lava Ultimate (LU), and Grandio blocs (GR). Stress distributions under axial and oblique loading were analyzed. The Weibull function was combined with the FEA results to predict long-term failure probability. The mechanical failure behavior of endocrowns manufactured with these materials was tested by using a universal testing machine. Load-to-failure was recorded, and fractured specimens were subjected to fractography. The data were analyzed by 1-way ANOVA and the post hoc Tukey test (α=.05). RESULTS: The models of GR and LU exhibited a more even stress distribution. The Weibull analysis revealed that 5 models performed in a similar manner under normal occlusal forces, while LU and VE models achieved the highest probabilities during clenching. The fracture loads of GR (3808 ±607 N) were significantly higher than those of other materials (P<.05). More favorable failure modes were observed in the GR and VE groups. Fractography showed a greater probability of compression curls and arrest lines in the endocrowns of VE, LU, and GR groups. CONCLUSIONS: When restoring endodontically treated teeth, endocrown fabricated with composite resin exhibited a more uniform stress distribution and higher fracture resistance. More evidence from long-term clinical studies is needed to verify this effect.

A Cross-Protective Vaccine Against 4b and 1/2b Listeria monocytogenes.

Listeria monocytogenes (Lm) is a foodborne zoonotic pathogen that causes listeriosis with a mortality rate of 20-30%. Serovar 4b and 1/2b isolates account for most of listeriosis outbreaks, however, no listeriosis vaccine is available for either prophylactic or therapeutic use. Here, we developed a triple-virulence-genes deletion vaccine strain, and evaluated its safety, immunogenicity, and cross-protective efficiency. The virulence of NTSNΔactA/plcB/orfX was reduced 794-folds compared with the parental strain. Additionally, it was completely eliminated in mice at day 7 post infection and no obvious pathological changes were observed in the organs of mice after prime-boost immunization for 23 days. These results proved that the safety of the Lm vaccine strain remarkably increased. More importantly, the NTSNΔactA/plcB/orfX strain stimulated higher anti-Listeriolysin O (LLO) antibodies, induced significantly higher expression of IFN-γ, TNF-α, IL-17, and IL-6 than the control group, and afforded 100% protection against serovar 4b and 1/2b challenges. Taken together, our research demonstrates that the triple-genes-deletion vaccine has high safety, can elicit strong Th1 type immune response, and affords efficient cross-protection against two serovar Lm strains. It is a promising vaccine for prevention of listeriosis.

Clinical efficacy of ceramic versus resin-based composite endocrowns in Chinese adults: study protocol for a randomized controlled trial.

BACKGROUND: Endocrown restoration is widely used to restore endodontically treated teeth. However, the clinical effects of different computer-aided design/computer-aided manufacturing (CAD/CAM) materials for endocrown restoration are not clear. The primary objective of this trial is to compare the clinical efficacy of resin-based bloc and ceramic endocrowns for restoring endodontically treated teeth. METHODS: The proposed resin-based bloc and ceramic endocrown assessment trial is a parallel group-designed randomized controlled trial. We will recruit 156 adults between 18 and 75 years old with a minimum of one such molar. The inclusion criteria were good oral hygiene habits, root apex of molar without evident damage, receipt of standard endodontic treatment, need for endocrown restoration, and only one endocrown restoration performed per patient. Patients participating in another study or those with systemic diseases, disabilities, or known allergies to used materials will be excluded. All patients will be randomized and restored with resin-based bloc and ceramic endocrown according to a random number table. Clinical evaluations will be performed at baseline and after treatment at 6, 12, and 24 months, in accordance with the modified Federation Dentaire Internationale (FDI) criteria, by two independent evaluators. The primary outcome is marginal adaptation; secondary outcomes include wear, tooth integrity, fracture of material and retention, marginal staining, and patient view. All data will be analyzed by an independent statistician. Signed rank-sum tests will be used for intragroup comparisons. Wilcoxon rank-sum tests will be used for intergroup comparisons. Hierarchical logistic regression will be used to adjust the baseline and other important indicators. DISCUSSION: This study will investigate endocrowns of two CAD/CAM materials for endodontically treated molars. The results may help clinicians choose the better CAD/CAM material option and explain to patients the advantages and disadvantages of these two materials with evidence-based support. For patients, the results may lead to improvement in long-term restoration. TRIAL REGISTRATION: ClinicalTrials.gov NCT04033380. Registered on 24 July 2019.

A hybrid sub-lineage of Listeria monocytogenes comprising hypervirulent isolates.

The foodborne pathogen Listeria monocytogenes (Lm) is a highly heterogeneous species and currently comprises of 4 evolutionarily distinct lineages. Here, we characterize isolates from severe ovine listeriosis outbreaks that represent a hybrid sub-lineage of the major lineage II (HSL-II) and serotype 4h. HSL-II isolates are highly virulent and exhibit higher organ colonization capacities than well-characterized hypervirulent strains of Lm in an orogastric mouse infection model. The isolates harbour both the Lm Pathogenicity Island (LIPI)-1 and a truncated LIPI-2 locus, encoding sphingomyelinase (SmcL), a virulence factor required for invasion and bacterial translocation from the gut, and other non-contiguous chromosomal segments from another pathogenic species, L. ivanovii. HSL-II isolates exhibit a unique wall teichoic acid (WTA) structure essential for resistance to antimicrobial peptides, bacterial invasion and virulence. The discovery of isolates harbouring pan-species virulence genes of the genus Listeria warrants global efforts to identify further hypervirulent lineages of Lm.

An essential role for hfq involved in biofilm formation and virulence in serotype 4b Listeria monocytogenes.

Regulator factor Hfq has been widely detected among both Gram-positive and Gram-negative bacteria; however, its role in Gram-positive bacteria is less well established and varies among species. In Listeria monocytogenes (Lm), an organism able to adapt to a range of environments and live both saprobiotic and parasitic lifestyles, the role of Hfq is not fully understood. Serotype 4b Listeria monocytogenes strains associated with the majority of listeriosis outbreak, while the function of hfq in serotype 4b strains still not referenced. Here, we constructed hfq deletion and reversion mutants of serotype 4b Lm NTSN and analysed the biological characteristics both in vitro and in vivo. The deletion of hfq resulted in a growth deficiency in medium containing 4.5% ethanol or 1% Triton X-100, and the growth of the mutant was significantly reduced at 4 °C. Furthermore, the hfq deletion dramatically decreased biofilm formation in BHI medium and gastric fluid medium, and reduced the invasion and replication rate into the Caco-2BBe cells and RAW264.7 cells. However, complementation restored the wild-type phenotype. Importantly, mouse infection experiments demonstrated that hfq played a more important role in the colonisation and virulence in serotype 4b strain Lm NTSN than in the serotype 1/2a strain Lm EGDe. Taken together, these results demonstrated that hfq is a novel factor associated with biofilm formation, and plays an essential role in the stress response and pathogenisis in serotype 4b strain Lm NTSN. Our data provide the basis for further research into the function of Hfq in serotype 4b Listeria monocytogenes.