Senescence-induced endothelial phenotypes underpin immune-mediated senescence surveillance.

Senescence is a stress-responsive tumor suppressor mechanism associated with expression of the senescence-associated secretory phenotype (SASP). Through the SASP, senescent cells trigger their own immune-mediated elimination, which if evaded leads to tumorigenesis. Senescent parenchymal cells are separated from circulating immunocytes by the endothelium, which is targeted by microenvironmental signaling. Here we show that SASP induces endothelial cell NF-κB activity and that SASP-induced endothelial expression of the canonical NF-κB component Rela underpins senescence surveillance. Using human liver sinusoidal endothelial cells (LSECs), we show that SASP-induced endothelial NF-κB activity regulates a conserved transcriptional program supporting immunocyte recruitment. Furthermore, oncogenic hepatocyte senescence drives murine LSEC NF-κB activity in vivo. Critically, we show two distinct endothelial pathways in senescence surveillance. First, endothelial-specific loss of Rela prevents development of Stat1-expressing CD4+ T lymphocytes. Second, the SASP up-regulates ICOSLG on LSECs, with the ICOS-ICOSLG axis contributing to senescence cell clearance. Our results show that the endothelium is a nonautonomous SASP target and an organizing center for immune-mediated senescence surveillance.

Role of oral microbiota in irreversible pulpitis - Current strategies and future perspectives.

Irreversible pulpitis is an inflammation of the tooth pulp caused by an opportunity-driven invasion of the pulp space by oral microbiota typically prevalent in the oral cavity. Microbial organisms are extensively recognised to be the fundamental cause of endodontic infections and treatment failures. Previously, bacterial species responsible for these infections were largely recognised using conventional microbial culture techniques, lending credence to the widely held belief that anaerobic Gram-negative bacteria frequently enter the pulp space and trigger endodontic infections. The advent of novel technologies grants the advantage of detecting and studying microbial populations via an amalgamation of the modern "Omics" techniques and meticulous bioinformatics analysis, additionally detecting the metatranscriptome, metaproteome and metabolome along with the metagenome. Amongst these analytical strategies, metagenomic analyses are essentially pragmatic for investigating the oral microbiome. Metagenomics favor not only assessment of microbial composition in diseased conditions, but also contributes to detection of novel, potentially pathogenic species inclusive of non-viable bacteria. The present review describes current knowledge of root canal microbiome, including its composition and functional attributes, the novel strategies available for detection of microbiome as well as challenges associated and provides some crucial pointers for areas of future research.

Acceptability of fully guided virtual implant planning software among dental undergraduate students.

BACKGROUND: Fully guided implant surgery as a technique is gaining popularity. It has been observed that use of surgical guides improves precision and predictability for dental implant placement. However, like any other newer technology, the acceptance of fully guided dental implant technology among users is based upon its perceived usability. This study aimed at evaluating the perception about using Virtual Implant Planning Software (VIPS) among undergraduate dental students at the university of Sharjah. METHODS: Ninety-Six dental surgery students from the University of Sharjah were included in the study. One week after the Virtual Implant Planning Software (Planmeca Romexis version 6.2 procedure, students were asked to complete a Combined technology acceptance model and the theory of planned behaviour (C-TAM TPB) questionnaire. Sixty-six students responded to the questionnaire. RESULTS: Cronbach's alpha surpassed 0.7 for perceived usefulness, perceived ease of use, perceived behavioral control, and subjective norm. Attitude and behavioural intention reported Cronbach's alpha values less than 0.7. Spearman's correlation coefficient was significant for all the constructs. Perceived ease of use explained 49%, 33%, and 42% of the variance of perceived usefulness (R2 = 0.49), attitude (R2 = 0.33), and perceived behavioral control (R2 = 0.42) respectively. Perceived usefulness explained 25%, 18%, and 23% of the variance of attitude (R2 = 0.25), behavioral intention (R2 = 0.18), and perceived behavioral control (R2 = 0.23) respectively. Attitude accounted for 25%, 33%, and 29% of the variance of behavioral intention (R2 = 0.25), perceived behavioral control (R = 0.33), and subjective norm (R = 0.29) respectively. CONCLUSION: The fully guided VIPS was acceptable by dental students specifically because of its usability. This makes VIPS a very effective tool for teaching implantology for dental students. VIPS also allows students to perform multiple repetitions of the implant planning procedure which enhances understanding and content retention.

Scavenger Receptors: Novel Roles in the Pathogenesis of Liver Inflammation and Cancer.

The scavenger receptor superfamily represents a highly diverse collection of evolutionarily-conserved receptors which are known to play key roles in host homeostasis, the most prominent of which is the clearance of unwanted endogenous macromolecules, such as oxidized low-density lipoproteins, from the systemic circulation. Members of this family have also been well characterized in their binding and internalization of a vast range of exogenous antigens and, consequently, are generally considered to be pattern recognition receptors, thus contributing to innate immunity. Several studies have implicated scavenger receptors in the pathophysiology of several inflammatory diseases, such as Alzheimer's and atherosclerosis. Hepatic resident cellular populations express a diverse complement of scavenger receptors in keeping with the liver's homeostatic functions, but there is gathering interest in the contribution of these receptors to hepatic inflammation and its complications. Here, we review the expression of scavenger receptors in the liver, their functionality in liver homeostasis, and their role in inflammatory liver disease and cancer.

Outcomes After Liver Transplantation With Incidental Cholangiocarcinoma.

Cholangiocarcinoma (CCA) is currently a contraindication to liver transplantation (LT) in the United Kingdom (UK). Incidental CCA occurs rarely in some patients undergoing LT. We report on retrospective outcomes of patients with incidental CCA from six UK LT centres. Cases were identified from pathology records. Data regarding tumour characteristics and post-transplant survival were collected. CCA was classified by TNM staging and anatomical location. 95 patients who underwent LT between 1988-2020 were identified. Median follow-up after LT was 2.1 years (14 days-18.6 years). Most patients were male (68.4%), median age at LT was 53 (IQR 46-62), and the majority had underlying PSC (61%). Overall median survival after LT was 4.4 years. Survival differed by tumour site: 1-, 3-, and 5-year estimated survival was 82.1%, 68.7%, and 57.1%, respectively, in intrahepatic CCA (n = 40) and 58.5%, 42.6%, and 30.2% in perihilar CCA (n = 42; p = 0.06). 1-, 3-, and 5-year estimated survival was 95.8%, 86.5%, and 80.6%, respectively, in pT1 tumours (28.2% of cohort), and 65.8%, 44.7%, and 31.1%, respectively, in pT2-4 (p = 0.018). Survival after LT for recipients with incidental CCA is inferior compared to usual outcomes for LT in the United Kingdom. LT for earlier stage CCA has similar survival to LT for hepatocellular cancer, and intrahepatic CCAs have better survival compared to perihilar CCAs. These observations may support LT for CCA in selected cases.

Comparison of clinical outcomes of non-surgical adjunctive use of the 980-nm diode laser to conventional periodontal flap procedure-a randomised controlled split-mouth clinical trial.

Lasers have been found to be effective as an adjunctive in periodontal therapy. The objective of the study was to compare the outcomes of non-surgical adjunctive use of 980-nm diode laser to that of conventional flap surgery in terms of clinical outcome measures and radiographic osseous defect depths. This study was designed as a double-blinded split-mouth randomised controlled clinical trial. A total of fifteen patients (30 quadrants) with generalised periodontitis stage III grade B were randomly assigned test group (diode laser) and control group (conventional flap surgery). Both test and control groups had 15 quadrants each. At 3 months, the probing pocket depth (PPD) was 3.46 ± 0.19 mm (test) and 4.20 ± 0.20 mm (control), with a statistically significant difference between the groups (p = 0.01). Similarly, at 6 months re-evaluation, PPD was 1.46 ± 0.12 mm (test) and 2.33 ± 0.13 mm (control) with a highly significant statistical difference (p = 0.001). Plaque index showed a difference between both test and control groups at baseline, 3 months and 6 months (p = 0.001) while gingival index, modified sulcular bleeding index and radiographic osseous defect depth showed a difference at 3 months and 6 months (p = 0.001). Within the study's limitations, 980-nm diode laser can result in significant improvements in the clinical and radiographic parameters and is very well tolerated by the subjects.

Clinical efficacy of green tea, aloe vera and chlorhexidine mouthwashes in the treatment of dental biofilm induced gingivitis: A multi-arm, double-blinded, randomized controlled clinical trial.

OBJECTIVE: This multi-arm, randomized, double-blinded, controlled clinical trial was designed to evaluate the clinical efficacy of 0.5% green tea (GT), 0.2% chlorhexidine (CHX) and aloe vera (AV) mouthwash as compared to the control (CNT) group (scaling and polishing alone with no mouthwash) in the management of dental biofilm induced gingivitis among 18-40-year-old patients. MATERIAL AND METHODS: Sixty patients with generalized dental biofilm-induced gingivitis were randomly allocated to four study groups (n = 15 each) for treatment, namely Group GT, Group CHX, Group AV and Group CNT after scaling and polishing were administered to all the patients. Plaque index (PI), gingival index (GI) and sulcular bleeding index (SBI) were recorded at baseline, 14th and 21st day. RESULTS: PI, GI and SBI at various time intervals (baseline, 14th and 21st day) showed high statistically significant differences within the group (p < 0.01). Among these, the maximum percentage change was found in the CHX group followed by GT when evaluated from baseline to 21st day. The least percentage change was found in the AV group for PI and GI while the CNT group showed the least percentage change for sulcular index when evaluated from 14th to 21st day. Inter-group results showed high statistically significant differences (p < 0.01) for PI and GI when evaluated between baseline to 14th day and baseline to 21st day. SBI also showed statistically significant differences (p < 0.05) when evaluated between baselines to 14th day and baseline to 21st day. CONCLUSION: Green tea mouthwash displayed a significant reduction in plaque index, gingival index and sulcular bleeding index. 0.5% green tea catechin has equivalent anti-plaque efficacy as 0.2% chlorhexidine gluconate and can be considered a potent alternative to prevent and treat gingival diseases.

Hepatocellular Carcinoma Demonstrates Heterogeneous Growth Patterns in a Multicenter Cohort of Patients With Cirrhosis.

BACKGROUND AND AIMS: There are limited data on hepatocellular carcinoma (HCC) growth patterns, particularly in Western cohorts, despite implications for surveillance, prognosis, and treatment. Our study's aim was to quantify tumor doubling time (TDT) and identify correlates associated with indolent and rapid growth. APPROACH AND RESULTS: We performed a retrospective multicenter cohort study of patients with cirrhosis diagnosed with HCC from 2008 to 2017 at six US and European health systems with two or more contrast-enhanced imaging studies performed ≥ 30 days apart prior to HCC treatment. Radiologists independently measured tumors in three dimensions to calculate TDT and specific growth rate (SGR). We used multivariable ordinal logistic regression to identify factors associated with indolent (TDT > 365 days) and rapid (TDT < 90 days) tumor growth. In the primary cohort (n = 242 patients from four centers), median TDT was 229 days (interquartile range [IQR], 89-627) and median SGR was 0.3% per day (IQR, 0.1%-0.8%). Over one-third (38%) of HCCs had indolent growth, 36.8% intermediate growth, and 25.2% rapid growth. In multivariable analysis, indolent growth was associated with larger tumor diameter (odds ratio [OR], 1.15, 95% confidence interval [CI], 1.03-1.30) and alpha-fetoprotein < 20 ng/mL (OR, 1.90; 95% CI, 1.12-3.21). Indolent growth was more common in nonviral than viral cirrhosis (50.9% versus 32.1%), particularly in patients with T1 HCC (OR, 3.41; 95% CI, 1.08-10.80). Median TDT (169 days; IQR 74-408 days) and SGR (0.4% per day) were similar in an independent cohort (n = 176 patients from two centers). CONCLUSIONS: In a large Western cohort of patients with HCC, we found heterogeneous tumor growth patterns, with one-fourth exhibiting rapid growth and over one-third having indolent growth. Better understanding different tumor growth patterns may facilitate a precision approach to prognostication and treatment.

Serum ceruloplasmin as cancer marker in oral pre-cancers and cancers.

BACKGROUND AND AIM: Oxidative stress leads to a compensatory increase in levels of serum ceruloplasmin in patients with such imbalances. Greater than normal serum ceruloplasmin levels are noticed in numerous cancers including the leukemias and Hodgkin's lymphoma. The purpose of the present study was to estimate and evaluate the efficacy of serum ceruloplasmin levels as a potential biomarker in the early detection of oral potentially malignant epithelial lesions (PMELs) including leukoplakia, oral submucous fibrosis (OSMF), and oral squamous cell carcinoma (OSCC) patients. MATERIALS AND METHODS: The present observational study was conducted over a period of 2 years wherein 100 subjects aged between 18 to 60 years were divided into four groups with Group A consisting of 25 healthy controls, Group B and C with 25 patients each, clinically diagnosed with oral leukoplakia and OSMF and Group D with 25 patients clinically diagnosed and histopathologically proven OSCC. The patients were subjected to incisional biopsy after routine hematological investigation while the same sera samples were used for analysis of serum ceruloplasmin levels. STATISTICAL ANALYSIS USED: Comparison of serum ceruloplasmin levels between the groups was performed using one way analysis of variance (one way ANOVA) test while P < 0.05 was considered statistically significant. RESULTS: The mean serum ceruloplasmin levels were found to be 43.19 ± 1.90mg/dl in subjects of group A, 47.68 ± 1.51mg/dl in group B, 47.74 ± 1.45mg/dl in group C and 47.73 ± 0.74mg/dl in group D. Using one-way ANOVA, statistically significant variations were found in the values of mean serum ceruloplasmin levels in subjects of the four groups (F-value = 59.58, P = 0.0001). CONCLUSIONS: The observations of the present study revealed that serum ceruloplasmin levels were found to be raised in all 3 study groups including oral leukoplakia, OSMF and OSCC as compared to the controls while the results were found to be statistically significant.

Three-dimensional qualitative and quantitative analyses of the effect of periradicular lesions on the outcome of regenerative endodontic procedures: A prospective clinical study.

OBJECTIVES: Qualitative and quantitative evaluation of the outcomes of regenerative endodontic procedure (REP) on human immature necrotic teeth with apical periodontitis using cone-beam computed tomography (CBCT) MATERIALS AND METHODS: Immature permanent teeth (n = 50) with necrotic pulp and periradicular pathosis were treated with a cell-homing concept-based REP. Following the procedure, a limited field-of-view CBCT scan was obtained. At each recall session (6, 12, 18 months), clinical tests were performed, and a digital periapical radiograph was taken. When significant radiographic changes were evident in the follow-up, a final CBCT scan was taken for qualitative and quantitative assessment. These initial and follow-up CBCT scans were assessed for quantification of changes in root length, pulp space diameter and periradicular lesion size. The data were statistically analysed using t test, one-way ANOVA, post hoc test and paired t test (P = 0.05). RESULTS: Of the teeth, 94.6% were clinically successful based on the lack or regression of signs and symptoms after 48 months follow-up. REP resulted in a statistically significant increase in root length, decrease in pulp space diameter and periradicular radiolucency (P < 0.05). CBCT images illustrated various patterns of root maturation including an increased thickening of the canal walls and continued root maturation (37.1%), continued root development with the apical foramen remaining open (57.1%), severe calcification (obliteration) of the canal space (2.9%) and hard tissue barrier formation in the canal space between the coronal plug and the root apex (2.9%). CONCLUSION: This study highlighted that the expected outcome of radiographic root development was less predictable when immature permanent teeth with periradicular pathosis were treated with REP. CLINICAL RELEVANCE: The size and extent (expansion/destruction of the cortical plate) of periradicular lesions and abscesses influence the outcome of REP. These factors must be taken into consideration during treatment planning.

The effect of concha bullosa and nasal septal deviation on palatal dimensions: a cone beam computed tomography study.

INTRODUCTION: Nasal septal deviation (NSD) and concha bullosa (CB) are associated with airway obstruction in mouth breathers. Mouth breathing is associated with alterations in maxillary growth and palatal architecture. The aim of our study was to determine the effect of the presence of CB and NSD on the dimensions of the hard palate using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: A retrospective study was conducted using CBCT scans of 200 study subjects. The study subjects were divided into four groups based on the presence of CB and NSD. Septal deviation angle (SDA), palatal interalveolar length (PIL), palatal depth (PD) and maxillopalatal arch angle (MPAA) were measured in the study groups. RESULTS: The presence of NSD and CB was associated with significant (p < 0.001) differences in the palatal dimensions of the study subjects. The PIL and MPA (p < 0.001) were significantly reduced (p < 0.001), whereas the PD was significantly increased (p < 0.001) in study subjects with NSD and CB. There was no significant change in the palatal dimensions between the unilateral and bilateral types of CB. Among the palatal dimensions, the PIL had the most significant association (R2 = 0.53) with SDA and CB. There was a significant correlation between the palatal dimensions and SDA when CB was present along with NSD. CONCLUSION: Based on the results of this study, it can be concluded that the presence of NSD and CB have a significant effect on the palatal dimensions and, therefore, they may be associated with skeletal malocclusion.

Analysis of inferior nasal turbinate width and concha bullosa in subjects with nasal septum deviation: a cone beam tomography study.

BACKGROUND: In individuals with nasal septal deviation (NSD), compensatory hypertrophy of the nasal turbinates occurs as a protective mechanism of the nasal passage from dry and cold air. NSD associated nasal turbinate hypertrophy is usually recurrent, requiring repetitive imaging. Therefore, a multiplanar imaging modality with a low radiation dose is best suited for long-term follow-up of this condition. This study aimed to evaluate the association of width of inferior turbinates and presence of concha bullosa with the degree of NSD using Cone beam computed tomography (CT). METHODS: The CBCT scans of 100 patients with NSD were selected as per convenience sampling and were evaluated by two maxillofacial radiologists. The width of the non-hypertrophied inferior turbinate (NHT) on the convex side of the NSD, and hypertrophic inferior turbinates (HT) on the concave side of the NSD were measured at three locations. The septal deviation angle (SDA) and the presence of concha bullosa (CB) were determined. RESULTS: A significant difference was observed in the anterior, middle, posterior, and mean widths between HT and NHT (p < 0.001). There was a significant difference in the widths of the HT and NHT among different types of NSD. A strong positive correlation (r = 0.71, p < 0.001) was found between SDA and the mean width of the HT. Age (P = 0.71) and gender (P = 0.65) had no significant difference among different types of NSD. Regression analysis revealed that the presence of CB (p = 0.01) and middle width of the HT (p < 0.001) are significant predictors of SDA and type of NSD. CONCLUSION: The results of the present study reveal that the middle width of the HT and the presence of CB influence the degree of NSD. The present study results recommend the use of CBCT as a substitutive low radiation dose imaging modality for evaluation of NSD, CB, and associated inferior turbinate hypertrophy.

Hypothermic Oxygenated Liver Perfusion (HOPE) Prevents Tumor Recurrence in Liver Transplantation From Donation After Circulatory Death.

OBJECTIVE: The aim of this study was to investigate tumor recurrence after liver transplantation for hepatocellular carcinoma (HCC), with and without hypothermic oxygenated liver perfusion (HOPE) before transplantation. PATIENTS AND METHODS: We analyzed all liver recipients with HCC, transplanted between January 2012 and September 2019 with donation after circulatory death (DCD) livers after previous end-ischemic HOPE-treatment (n = 70, Center A). Tumor parameters and key confounders were compared to consecutive recipients with HCC, transplanted during the same observation period with an unperfused DBD liver (n = 70). In a next step, we analyzed unperfused DCD (n = 70) and DBD liver recipients (n = 70), transplanted for HCC at an external center (Center B). RESULTS: Tumor parameters were not significantly different between HOPE-treated DCD and unperfused DBD liver recipients at Center A. One-third of patients were outside established tumor thresholds, for example, Milan criteria, in both groups. Despite no difference in tumor load, we found a 4-fold higher tumor recurrence rate in unperfused DBD livers (25.7%, 18/70), compared to only 5.7% (n = 4/70) recipients with tumor recurrence in the HOPE-treated DCD cohort (P = 0.002) in Center A. The tumor recurrence rate was also twice higher in unperfused DCD and DBD recipients at the external Center B, despite significant less cases outside Milan. HOPE-treatment of DCD livers resulted therefore in a 5-year tumor-free survival of 92% in HCC recipients, compared to 73%, 82.7%, and 81.2% in patients receiving unperfused DBD or DCD livers, from both centers. CONCLUSION: We suggest that a simple machine liver perfusion approach appears advantageous to protect from HCC recurrence after liver transplantation, despite extended tumor criteria.

Novel Targets in the Immune Microenvironment of the Hepatic Sinusoids for Treating Liver Diseases.

Immune dysregulation and accumulation of leukocytes is a hallmark of adult chronic liver diseases. Progressive hepatic inflammation can lead to fibrosis and cirrhosis with a high risk of liver failure or hepatocellular cancer (HCC). Recent advances have been made in the treatment of liver disease including the development of highly effective antiviral therapy for hepatitis C and the potential of immunotherapy for HCC. Despite this, the majority of other chronic liver diseases including alcoholic liver disease, fatty liver disease, and cholestatic diseases do not respond to conventional anti-inflammatory therapies. Recent studies defining the organ-specific properties that contribute to resident immune activation and immune cell recruitment from the circulation in these conditions have identified novel hepatic inflammatory pathways, which are now being targeted in clinical trials. Further understanding of how the immune microenvironment is regulated within the liver and how disease-specific mechanisms alter this process will hopefully lead to combination therapies to prevent aberrant inflammation and also promote fibrosis resolution. In this review, we focus on the advances that have been made in identifying key components of the inflammatory pathway including the recognition of danger signals, the recruitment and retention of lymphocytes from the circulation, and the pathways that promote resolution.

Clinical implications of heterogeneity in PD-L1 immunohistochemical detection in hepatocellular carcinoma: the Blueprint-HCC study.

Programmed cell death ligand-1 immunohistochemical detection (PD-L1 IHC) is a putative predictor of response to PD-1/PD-L1-targeted checkpoint inhibitors. However, there is no gold standard assay in hepatocellular carcinoma (HCC). We evaluated 5 PD-L1 IHC assay platforms (E1LN3, 28-8, 22c3, SP263 and SP142) in 100 HCCs reporting PD-L1 expression in malignant (M) and tumour-infiltrating immune cells (TICs) and non-tumorous cirrhotic tissues (NTICs). We found substantial inter-assay heterogeneity in detecting PD-L1 expression in M (R2 = 0.080-0.921), TICs (Cohen's κ = 0.175-0.396) and NTICs (κ = 0.004-0.505). Such diversity may impact on the reliability and reproducibility of PD-L1 IHC assays as a predictor of response to immune checkpoint inhibitors.

Stabilin-1 expression defines a subset of macrophages that mediate tissue homeostasis and prevent fibrosis in chronic liver injury.

Macrophages are key regulators of fibrosis development and resolution. Elucidating the mechanisms by which they mediate this process is crucial for establishing their therapeutic potential. Here, we use experimental models of liver fibrosis to show that deficiency of the scavenger receptor, stabilin-1, exacerbates fibrosis and delays resolution during the recovery phase. We detected a subset of stabilin-1(+) macrophages that were induced at sites of cellular injury close to the hepatic scar in mouse models of liver fibrosis and in human liver disease. Stabilin-1 deficiency abrogated malondialdehyde-LDL (MDA-LDL) uptake by hepatic macrophages and was associated with excess collagen III deposition. Mechanistically, the lack of stabilin-1 led to elevated intrahepatic levels of the profibrogenic chemokine CCL3 and an increase in GFAP(+) fibrogenic cells. Stabilin-1(-/-) macrophages demonstrated a proinflammatory phenotype during liver injury and the normal induction of Ly6C(lo) monocytes during resolution was absent in stabilin-1 knockouts leading to persistence of fibrosis. Human stabilin-1(+) monocytes efficiently internalized MDA-LDL and this suppressed their ability to secrete CCL3, suggesting that loss of stabilin-1 removes a brake to CCL3 secretion. Experiments with cell-lineage-specific knockouts revealed that stabilin-1 expression in myeloid cells is required for the induction of this subset of macrophages and that increased fibrosis occurs in their absence. This study demonstrates a previously unidentified regulatory pathway in fibrogenesis in which a macrophage scavenger receptor protects against organ fibrosis by removing fibrogenic products of lipid peroxidation. Thus, stabilin-1(+) macrophages shape the tissue microenvironment during liver injury and healing.

Microleakage Evaluation of an Alkasite Restorative Material: An In Vitro Dye Penetration Study.

AIM: The aim of the present in vitro study was to comparatively evaluate the microleakage in three different esthetic restorative materials in class I cavities using the dye penetration technique. MATERIALS AND METHODS: Class I cavities were prepared on 24 human maxillary premolar teeth. The teeth were randomly divided into four groups of six samples each. Group I: Cention-N without adhesive (Ivoclar Vivadent, India), group II: Cention with adhesive (Ivoclar Vivadent, India), group III: type IX glass ionomer cement (Fuji), group IV: posterior composite (3M ESPE). The specimens were polished, subjected to thermocycling, and suspended in methylene blue dye for 24 hours. The teeth were sectioned longitudinally and the extent of microleakage was evaluated using the stereomicroscope. RESULTS: The results were subjected to statistical analysis using the Pearson's Chi-square test and the interobserver variability was assessed by the Kappa test for interobserver variability. The analysis showed statistically significant results among the groups. Although, Cention N with adhesive showed the least microleakage followed by Cention N without adhesive. CONCLUSION: All the materials tested were unable to completely eliminate microleakage in class I cavities. However, the newer alkasite material Cention N proved to have the least microleakage among all groups. CLINICAL SIGNIFICANCE: According to the present study, Cention N, a newer alkasite restorative material, demonstrated promising results with the least microleakage in comparison with posterior resin composites and glass ionomer cements. How to cite this article: Kini A, Shetty S, Bhat R, et al. Microleakage Evaluation of an Alkasite Restorative Material: An In Vitro Dye Penetration Study. J Contemp Dent Pract 2019;20(11):1315-1318.

Human liver sinusoidal endothelial cells promote intracellular crawling of lymphocytes during recruitment: A new step in migration.

The recruitment of lymphocytes via the hepatic sinusoidal channels and positioning within liver tissue is a critical event in the development and persistence of chronic inflammatory liver diseases. The hepatic sinusoid is a unique vascular bed lined by hepatic sinusoidal endothelial cells (HSECs), a functionally and phenotypically distinct subpopulation of endothelial cells. Using flow-based adhesion assays to study the migration of lymphocytes across primary human HSECs, we found that lymphocytes enter into HSECs, confirmed by electron microscopy demonstrating clear intracellular localization of lymphocytes in vitro and by studies in human liver tissues. Stimulation by interferon-γ increased intracellular localization of lymphocytes within HSECs. Furthermore, using confocal imaging and time-lapse recordings, we demonstrated "intracellular crawling" of lymphocytes entering into one endothelial cell from another. This required the expression of intracellular adhesion molecule-1 and stabilin-1 and was facilitated by the junctional complexes between HSECs. CONCLUSION: Lymphocyte migration is facilitated by the unique structure of HSECs. Intracellular crawling may contribute to optimal lymphocyte positioning in liver tissue during chronic hepatitis. (Hepatology 2017;65:294-309).

CD151 supports VCAM-1-mediated lymphocyte adhesion to liver endothelium and is upregulated in chronic liver disease and hepatocellular carcinoma.

CD151, a member of the tetraspanin family of receptors, is a lateral organizer and modulator of activity of several families of transmembrane proteins. It has been implicated in the development and progression of several cancers, but its role in chronic inflammatory disease is less well understood. Here we show that CD151 is upregulated by distinct microenvironmental signals in a range of chronic inflammatory liver diseases and in primary liver cancer, in which it supports lymphocyte recruitment. CD151 was highly expressed in endothelial cells of the hepatic sinusoids and neovessels developing in fibrotic septa and tumor margins. Primary cultures of human hepatic sinusoidal endothelial cells (HSECs) expressed CD151 at the cell membrane and in intracellular vesicles. CD151 was upregulated by VEGF and HepG2 conditioned media but not by proinflammatory cytokines. Confocal microscopy confirmed that CD151 colocalized with the endothelial adhesion molecule/immunoglobulin superfamily member, VCAM-1. Functional flow-based adhesion assays with primary human lymphocytes and HSECs demonstrated a 40% reduction of lymphocyte adhesion with CD151 blockade. Inhibition of lymphocyte adhesion was similar between VCAM-1 blockade and a combination of CD151/VCAM-1 blockade, suggesting a collaborative role between the two receptors. These studies demonstrate that CD151 is upregulated within the liver during chronic inflammation, where it supports lymphocyte recruitment via liver endothelium. We propose that CD151 regulates the activity of VCAM-1 during lymphocyte recruitment to the human liver and could be a novel anti-inflammatory target in chronic liver disease and hepatocellular cancer prevention.NEW & NOTEWORTHY Chronic hepatitis is characterized by lymphocyte accumulation in liver tissue, which drives fibrosis and carcinogenesis. Here, we demonstrate for the first time that the tetraspanin CD151 supports lymphocyte adhesion to liver endothelium. We show that CD151 is upregulated in chronic liver disease and hepatocellular carcinoma (HCC) and is regulated on endothelium by tissue remodeling and procarcinogenic factors. These regulatory and functional studies identify CD151 as a potential therapeutic target to treat liver fibrosis and HCC.