The adult pineal gland is composed of pinealocytes, astrocytes, microglia, and other interstitial cells that have been described in detail. However, factors that contribute to pineal development have ...not been fully elucidated, nor have pineal cell lineages been well characterized. We applied systematic double, triple and quadruple labeling of cell-specific markers on prenatal, postnatal and mature rat pineal gland tissue combined with confocal microscopy to provide a comprehensive view of the cellular dynamics and cell lineages that contribute to pineal gland development. The pineal gland begins as an evagination of neuroepithelium in the roof of the third ventricle. The pineal primordium initially consists of radially aligned Pax6+ precursor cells that express vimentin and divide at the ventricular lumen. After the tubular neuroepithelium fuses, the distribution of Pax6+ cells transitions to include rosette-like structures and later, dispersed cells. In the developing gland all dividing cells express Pax6, indicating that Pax6+ precursor cells generate pinealocytes and some interstitial cells. The density of Pax6+ cells decreases across pineal development as a result of cellular differentiation and microglial phagocytosis, but Pax6+ cells remain in the adult gland as a distinct population. Microglial colonization begins after pineal recess formation. Microglial phagocytosis of Pax6+ cells is not common at early stages but increases as microglia colonize the gland. In the postnatal gland microglia affiliate with Tuj1+ nerve fibers, IB4+ blood vessels, and Pax6+ cells. We demonstrate that microglia engulf Pax6+ cells, nerve fibers, and blood vessel-related elements, but not pinealocytes. We conclude that microglia play a role in pineal gland formation and homeostasis by regulating the precursor cell population, remodeling blood vessels and pruning sympathetic nerve fibers.
SARS-CoV-2 virus infects children but, contrary to other respiratory viruses, children tend to be asymptomatic or to have less symptoms than adults and are rarely the index case in household ...transmission chains. The aim of this study was to compare the relative viral loads in nasopharyngeal samples from children aged 0 to 17 years with those of an adult population. We selected 126 positive nasopharyngeal samples from children. Relative viral loads were measured by RT-PCR and the comparative ∆Ct method using primers and probes targeting the viral gene E and the human RNAseP. Relative viral loads were significantly lower in the children population than in the adult one.
Are Denture Adhesives Safe for Oral Cells? López‐García, Sergio; Pecci‐Lloret, María P.; García‐Bernal, David ...
Journal of prosthodontics,
January 2021, 2021-Jan, 2021-01-00, 20210101, Letnik:
30, Številka:
1
Journal Article
Recenzirano
Purpose
To compare the cytotoxicity of six commercially available denture adhesives on human gingival cells: Poligrip Flavour Free Fixative Cream, Fixodent Pro Duo Protection, Novafix cream, ...FittyDent, Polident Total Action, and Fixodent Pro Plus Duo Protection.
Material and Methods
Eluates of denture adhesives were brought into contact with human gingival cells and compared to untreated cells (w/o any dental adhesive elute). Cell toxicity was assessed by measuring cell viability (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide) (MTT) assays), cell morphology (immunofluorescence assays), induction of apoptosis/necrosis and production of reactive oxygen species (ROS) (flow cytometry assays). In addition, the pH of each sample was determined. Data were analyzed using one‐way analysis of variance (ANOVA) followed by Dunnett's multiple comparisons test.
Results
All denture adhesives tested led to a reduction in pH, especially Fixodent Pro Duo Protection and Fixodent Pro Plus Duo Protection. The cell viability assays showed that Fixodent Pro Duo Protection (1:1 72 hours, p = 3.04 × 10−6; 1:2 72 hours, p = 2.07 × 10−6; 1:4 72 hours, p = 2.04 × 10−6) and Fixodent Pro Plus Duo Protection (1:1 72 hours, p = 2.01 × 10−6; 1:2 72 hours, p = 3.03 × 10−6; 1:4 72 hours, p = 2.02 × 10−6) significantly decreased cell viability at all dilutions. Compared to the control group and the rest of the adhesives, Poligrip Flavour Free Fixative Cream (PFF 1:1 72 hours, p = 2.24 × 10−6; 1:2 72 hours, p = 2.44 × 10−6; 1:4 72 hours, p = 2.04 × 10−6) showed a significantly higher cell viability score at all dilutions. Fixodent Pro Duo Protection and Fixodent Pro Plus Duo Protection, both adhesives containing zinc salts in their composition, were responsible for necrosis, and the number of cells was much reduced, with aberrant morphology and pyknotic nucleus. Finally, Fixodent (1:2, p = 2.04 × 10−6, 1:4, p = 0.00036; 1:2, p = 8.82 × 10−6, 1:4, p = 2.30 × 10−6) products significantly promoted ROS production in gingival cells.
Conclusions
The results suggest that denture adhesives containing zinc in their composition could be responsible of the decrease of cell viability, ROS production, aberrant cell morphology, and induction of apoptosis and cell death. However, other possible additional cytotoxic factors must be considered. Thus, more studies are necessary to confirm this hypothesis.
The aim of this study was to investigate the cytocompatibility and mineralization potential of two premixed hydraulic endodontic sealers compared with an epoxy resin-based root canal sealer. The ...cellular responses and mineralization capacity were studied in human periodontal ligament stem cells (hPDLSCs) that were exposed to premixed hydraulic sealers, Bio-C Sealer (Angelus, Londrína, PR, Brazil), TotalFill BC Sealer (FKG Dentaire SA, La-Chaux-de-fonds, Switzerland) and an epoxy resin-based material, AH Plus (Dentsply De Trey, Konstanz, Germany). Non-exposed cultures served as the control. The endodontic sealers were assessed using scanning electron microscopy (SEM) and energy dispersive X-ray microanalysis (EDX). Statistical analyses were done using Analisis of Variance (ANOVA), with Bonferroni adjusted pairwise comparison (p = 0.05). AH Plus reduced cell viability and cell migration, whereas increased cell viability and cell migration were observed in the Bio-C Sealer and the TotalFill BC Sealer (p < 0.05). The lowest cell attachment and spreading were observed for all concentrations of AH Plus, whereas the highest were observed for TotalFill BC Sealer. At the end of 21 days, only the Bio-C Sealer and the TotalFill BC Sealer supported matrix mineralization (p < 0.05). Additionally, SEM-EDX revealed high content of calcium, oxygen, and silicon in the Bio-C Sealer and the TotalFill BC Sealer. Based on the results from this study, Bio-C Sealer and TotalFill BC Sealer demonstrated better cytocompatibility in terms of cell viability, migration, cell morphology, cell attachment, and mineralization capacity than AH Plus.
To describe short-term and 5-year rates of mortality and poor outcome in patients with spontaneous aneurysmal subarachnoid hemorrhage (aSAH) who received repair treatment.
In this prospective ...observational study, mortality and poor outcome (modified Rankin Scale score 3-6) were analyzed in 311 patients with aSAH at 3 months, 1 year, and 5 years follow-up. Sensitivity analysis was performed according to treatment modality. In-hospital and 5-year complications were analyzed.
Of 476 consecutive patients with spontaneous subarachnoid hemorrhage, 347 patients (72.9%) had aSAH. Of these, 311 (89.6%) were treated (242 endovascular, 69 neurosurgical), with a mean follow-up of 43.4 months (range, 1 to 145). Three-month, 1-year, and 5-year mortality was 18.4%, 22.9%, and 29.0%, and poor outcome was observed in 42.3%, 36.0%, and 36.0%, respectively. Adjusted poor outcome was lower in endovascular than in neurosurgical treatment at 3 months (odds ratio OR 0.36 95% confidence interval CI 0.18-0.74), with an absolute difference of 15.8% (number needed to treat = 6.3), and at 1 year (OR = 0.40 95% CI 0.20-0.81), with an absolute difference of 15.9% (number needed to treat = 6.3). Complications did not differ between the 2 procedures. However, mechanical ventilation was less frequent with the endovascular technique (OR 0.67 95% CI 0.54-0.84).
Patients with aSAH treated according to current guidelines had a short-term mortality of 18.4% and 5-year mortality of 29%. The majority (64.0%) of patients remained alive without disabilities at 5-year follow-up. Patients prioritized to endovascular treatment had better outcomes than those referred to neurosurgery because endovascular coiling was not feasible.
We have determined the effects of the IGF-1R tyrosine kinase inhibitors BMS-754807 (BMS) and OSI-906 (OSI) on cell proliferation and cell-cycle phase distribution in human colon, pancreatic ...carcinoma, and glioblastoma cell lines and primary cultures. IGF-1R signaling was blocked by BMS and OSI at equivalent doses, although both inhibitors exhibited differential antiproliferative effects. In all pancreatic carcinoma cell lines tested, BMS exerted a strong antiproliferative effect, whereas OSI had a minimal effect. Similar results were obtained on glioblastoma primary cultures, where HGUE-GB-15, -16 and -17 displayed resistance to OSI effects, whereas they were inhibited in their proliferation by BMS. Differential effects of BMS and OSI were also observed in colon carcinoma cell lines. Both inhibitors also showed different effects on cell cycle phase distribution, BMS induced G
/M arrest followed by cell death, while OSI induced G
arrest with no cell death. Both inhibitors also showed different effects on other protein kinases activities. Taken together, our results are indicative that BMS mainly acts through off-target effects exerted on other protein kinases. Given that BMS exhibits a potent antiproliferative effect, we believe that this compound could be useful for the treatment of different types of tumors independently of their IGF-1R activation status.
Current recommendations for echocardiographic assessment of diastolic function (2016 guidelines of the American Society of Echocardiography (ASE)/European Association of Cardiovascular Imaging ...(EACVI) in patients with metabolic syndrome and overweight/obesity result in a significant number of patients with indeterminate diastolic dysfunction (LVDD). The aim of this article is to study whether the use of the left atrial strain criterion (LALS) reduces the number of indeterminate patients.
229 patients were studied with a complete echocardiographic study that included left ventricular longitudinal strain (LVLS) analysis, LALS and a maximal ergospirometry test with assessment of oxygen uptake (VO2max).
The mean age was 65 ± 5 years, 153 (67%) males, with a mean EF of 60 ± 5%. The mean LVLS was −19.4 ± 2% and the LALS Reservoir was 23.8 ± 7%. There were 140 patients who did not meet LVDD criteria and 82 who did meet the indeterminate LVDD criterion. When the left atrial volume index (LAVI) >34 ml/m2 criterion was replaced in the 2016 ASE/EACVI algorithm by LALS Reservoir ≤20%, the number of indeterminate patients was reduced from 36% to 23% (p < 0.001) at the expense of increasing normal studies (61% and 74%). Adding the LALS Reservoir criterion ≤23% in the 82 patients of the indeterminate group resulted in two groups with a different VO2max (11.6 ± 3 and 18 ± 5 ml/kg/min, p:0.081).
This study confirms the low prevalence of diastolic dysfunction in overweight/obese patients with metabolic syndrome. Adding left atrial strain criterion to the current recommendations significantly reduces the number of indeterminate patients by reclassifying them as normal.
•Diastolic dysfunction is often associated with metabolic syndrome and overweight/obesity.•Guidelines for echocardiographic diagnosis of diastolic dysfunction show a high proportion of indeterminate studies.•It explores the usefulness of longitudinal left atrial strain for echocardiographic assessment of diastolic function.•The use of this parameter reduces the number of indeterminate patients when applying current recommendations.
•3D printing resins have shown similar biological effects to conventional ones.•Freeprint Splint was the most cytotoxic on human gingival fibroblasts of the four dental resin studied.•The results ...obtained in this study suggest that hGFs showed better in vitro biocompatibility after exposition to Keysplint Soft, being the most eligible dental resin for occlusal devices.
: To assess and compare the in vitro biocompatibility of new resins (Keysplint Soft (Keystone Industries), NextDent Ortho Rigid (3D System), and Freeprint Splint (Detax)) and traditional resins (Orthocryl (Dentaurum)) for dental splint.
: Standardized discs (n=40) and 1:1, 1:2, and 1:4 extracts of the tested materials were prepared. Human gingival fibroblasts (hGFs) were isolated from gingival tissues. Different biological tests were carried out, including MTT assays to assess cell metabolic activity, cell migration assays, cell cytoskeleton staining, cell apoptosis, generation of intracellular reactive oxygen species (ROS), and scanning electron microscopy (SEM). Statistical analyses were performed using one-way ANOVA and Tukey's post hoc test (p<0.05).
: MTT experiments showed that Freeprint Splint significantly reduces the hGF metabolic activity (***p<0.001), whereas SEM analysis showed almost no cells adhered on its surface. Cell migration was significantly lower after exposure to undiluted extracts of Freeprint Splint at 48 and 72h (***p<0.001). Cell cytoskeleton staining assays showed fewer attached cells in 1:1 and 1:2 dilutions of Freeprint Splint. Annexin-V and 7-AAD staining assays showed that only cells exposed to Keysplint Soft extracts displayed similar cell viability to the control group. Finally, ROS levels detected in undiluted extracts of all resins were significantly enhanced compared to the control group (***p<0.001).
: The 3D-printed resins and the conventional dental resin showed a similar biocompatibility, except for Freeprint Splint, which was the most cytotoxic on hGFs.
: 3D printing has been on the rise in recent years and its use in daily clinical practice is expanding over time. Two of the three 3D-printed resins tested in this study performed as well in the cytotoxicity tests as the conventional one, supporting their use, but caution and further testing are required
Microglial cells are one of the interstitial elements of the pineal gland (PG). We recently reported the pattern of microglia colonization and activation, and microglia‐Pax6+ cell interactions during ...normal pineal ontogeny. Here, we describe the dynamics of microglia‐Pax6+ cell associations and interactions after surgical or pharmacological manipulation. In adult rats, the superior cervical ganglia (SCG) were exposed, and either bilaterally excised (SCGx) or decentralized (SCGd). In the SCGx PGs, the density of Iba1+ microglia increased after surgery and returned to sham baseline levels 13 days later. Pineal microglia also responded to SCGd, a more subtle denervation. The number of clustered Iba1+/PCNA+/ED1+ microglia was higher 4 days after both surgeries compared to the sham‐operated group. However, the number of Pax6+/PCNA− cells and the percentage of Pax6+ cells contacted by and/or phagocytosed by microglia increased significantly only after SCGx. Separate groups of rats were treated with either bacterial lipopolysaccharides (LPS) or doxycycline (DOX) to activate or inhibit pineal microglia, respectively. Peripheral LPS administration caused an increase in the number of clustered Iba1+/PCNA+/ED1+ microglial cells, and in the percentage of Pax6+ cells associated with and/or engulfed by microglia. In the LPS‐treated PGs, we also noted an increase in the number of PCNA+ cells that were Iba1− within the microglial cell clusters. The density of Pax6+ cells did not change after LPS treatment. DOX administration did not influence the parameters analyzed. These data suggest that pineal microglia are highly receptive cells capable of rapidly responding in a differential manner to surgical and pharmacological stimuli.
Pineal Iba1+/PCNA+/ED1+ microglia are constantly ‘activated’ cells able to sense and differentially respond to surgical and pharmacological stimuli. Here, we show how bilateral superior cervical ganglionectomy (SCGx) and decentralization (SCGd), and peripheral administration of bacterial lipopolysaccharides (LPS) and doxycycline (DOX) impact in a stimulus‐specific manner on microglia, Pax6+ cells, and associations and interactions between both cell types within the pineal gland. We propose the pineal gland as an attractive model to study phagocyte renewal and the interface between the immune system and the central nervous system.
Biocompatibility is an essential property for any vital pulp material that may interact with the dental pulp tissues. Accordingly, this study aimed to compare the chemical composition and ...ultrastructural morphology of Biodentine (Septodont, Saint Maur-des-Fosses, France), ProRoot MTA (Dentsply Tulsa Dental Specialties, Johnson City, TN, USA), and Bio-C Repair (Angelus, Londrina, PR, Brazil), as well as their biological effects on human dental pulp cells. Chemical element characterization of the materials was undertaken using scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX). The cytotoxicity was assessed by analyzing the cell viability (MTT assay), cell morphology (immunofluorescence assay), and cell attachment (flow cytometry assay). The results were statistically analyzed using ANOVA and Tukey's test (
< 0.05). EDX revealed that ProRoot MTA and Biodentine were mostly composed of calcium, carbon, and oxygen (among others), whereas Bio-C Repair evidenced a low concentration of calcium and the highest concentration of zirconium. SEM showed adequate attachment of human dental pulp cells (hDPCS) to vital pulp materials and cytoskeletal alterations were not observed in the presence of material eluates. Remarkably, the undiluted Biodentine group showed higher viability than the control group cells (without eluates) at 24 h, 48 h, and 72 h (
< 0.001). Based on the evidence derived from an in vitro cellular study, it was concluded that Bio-C Repair showed excellent cytocompatibility that was similar to Biodentine and ProRoot MTA.
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