Curcumin and quercetin co-encapsulated in nanoemulsions for nasal administration: A promising therapeutic and prophylactic treatment for viral respiratory infections.

One of the most frequent causes of respiratory infections are viruses. Viruses reaching the airways can be absorbed by the human body through the respiratory mucosa and mainly infect lung cells. Several viral infections are not yet curable, such as coronavirus-2 (SARS-CoV-2). Furthermore, the side effect of synthetic antiviral drugs and reduced efficacy against resistant variants have reinforced the search for alternative and effective treatment options, such as plant-derived antiviral molecules. Curcumin (CUR) and quercetin (QUE) are two natural compounds that have been widely studied for their health benefits, such as antiviral and anti-inflammatory activity. However, poor oral bioavailability limits the clinical applications of these natural compounds. In this work, nanoemulsions (NE) co-encapsulating CUR and QUE designed for nasal administration were developed as promising prophylactic and therapeutic treatments for viral respiratory infections. The NEs were prepared by high-pressure homogenization combined with the phase inversion temperature technique and evaluated for their physical and chemical characteristics. In vitro assays were performed to evaluate the nanoemulsion retention into the porcine nasal mucosa. In addition, the CUR and QUE-loaded NE antiviral activity was tested against a murine ß-COV, namely MHV-3. The results evidenced that CUR and QUE loaded NE had a particle size of 400 nm and retention in the porcine nasal mucosa. The antiviral activity of the NEs showed a percentage of inhibition of around 99 %, indicating that the developed NEs has interesting properties as a therapeutic and prophylactic treatment against viral respiratory infections.

MiRNAs that target amyloid precursor protein processing machinery in extracellular vesicles and particles derived from oral squamous cells carcinoma.

BACKGROUND: Considering that microRNAs (miRNAs), extracellular vesicles and particles (EVPs) and the amyloid precursor protein (APP) processing have been shown to be altered in oral squamous cells carcinoma (OSCC), it is possible that miRNAs that target APP processing pathways in EVPs are impacted in tumor cells. Our aim was to evaluate miRNAs that target APP itself or disintegrin and metalloproteinase domain 10 (ADAM10), which generate a trophic compound, sAPPα, in EVPs derived from OSCC cell lines, an aggressive and non-invasive, compared to normal keratinocytes. METHODS: We used two OSCC cell lines, an aggressive human oral squamous cell carcinoma cell line (SCC09) and a less aggressive cell line (CAL27) compared with a keratinocyte lineage (HaCaT). Cells were maintained in cell media, from which we isolated EVPs. EVPs were evaluated regarding their size and concentration using Nanotracking Analysis. We measured the levels of miRNAs which had as potential downstream target APP or ADAM10, specifically miR-20a-5p, miR-103a-3p, miR-424-5p, miR-92b-3p, miR-31-5p, and miR-93-5. RESULTS: There were no differences on size distributions and concentration of isolated EVPs. OSCC cell lines-derived EVPs miR-20a-5p, miR-92b-3p, and miR-93-5p were upregulated in comparison to HaCaT-derived EVPs; while miR-31-5p was reduced in EVPs obtained from CAL27 cells. CONCLUSION: Our results indicate changes in miRNAs that target APP machinery processing in EVPs derived from OSCC cell lines of different aggressiveness, which may be involved with abnormal miRNA expression in OSCC tissue and/or releasing tumor suppressor miRNA.

Cell contractility drives mechanical memory of oral squamous cell carcinoma.

Matrix stiffening is ubiquitous in solid tumors and can direct epithelial-mesenchymal transition (EMT) and cancer cell migration. Stiffened niche can even cause poorly invasive oral squamous cell carcinoma (OSCC) cell lines to acquire a less adherent, more migratory phenotype, but mechanisms and durability of this acquired "mechanical memory" are unclear. Here, we observed that contractility and its downstream signals could underlie memory acquisition; invasive SSC25 cells overexpress myosin II (vs. noninvasive Cal27 cells) consistent with OSCC. However, prolonged exposure of Cal27 cells to a stiff niche or contractile agonists up-regulated myosin and EMT markers and enabled them to migrate as fast as SCC25 cells, which persisted even when the niche softened and indicated "memory" of their prior niche. Stiffness-mediated mesenchymal phenotype acquisition required AKT signaling and was also observed in patient samples, whereas phenotype recall on soft substrates required focal adhesion kinase (FAK) activity. Phenotype durability was further observed in transcriptomic differences between preconditioned Cal27 cells cultured without or with FAK or AKT antagonists, and such transcriptional differences corresponded to discrepant patient outcomes. These data suggest that mechanical memory, mediated by contractility via distinct kinase signaling, may be necessary for OSCC to disseminate.

Cytotoxic, Migration, and Angiogenic Effects of Photodynamic Therapy and Photobiomodulation Associated with a Revascularization Protocol.

INTRODUCTION: This study evaluated photodynamic therapy (PDT) and photobiomodulation therapy (PBM) as adjuncts to pulp revascularization using cultures of apical papilla cells (APCs) and endothelial cells (HUVECs). METHODS: The root canal and apical foramen of 2 mandibular first premolars were enlarged to simulate immature teeth. The canal of 1 tooth was filled with 1 mL 0.005% methylene blue (MB). After that, the canals of both teeth were irrigated with 20 mL 1.5% sodium hypochlorite (NaOCl) and 20 mL 17% EDTA. The resulting solutions were diluted in cell culture media at a concentration of 0.5% (0.5% MB + NaOCl + EDTA and 0.5% NaOCl + EDTA). After PDT (0.5% MB + NaOCl + EDTA + PDT) and PBM (0.5% NaOCl + EDTA + PBM) applications, the effects were evaluated to determine cytotoxicity, polarity index, APC migration, and HUVEC sprouting, and results were compared with those of their controls (solutions without laser application). Cell culture media (CT) was also used as a control. Data were analyzed using 1-way analysis of variance and the Tukey post hoc test (P ≤ .05). RESULTS: PDT and PBM promoted greater APC viability than their controls, and PDT had greater cell viability than CT (P < .05). All protocols reduced APC migration when compared with CT (P < .05). HUVEC sprouts grown out of spheroids in PBM had a greater ratio area than their control (P ≤ .01), and the PDT ratio of the spheroid area was similar to that of its control (P > .05). CONCLUSIONS: PBM and PDT seem to be potentially effective adjuncts to revascularization in nonvital immature teeth.

Cytotoxic, migration, and angiogenic effects of intracanal irrigants on cells involved in revascularization of immature teeth.

OBJECTIVE: To evaluate protocols of root canal irrigation and dentin pretreatment in a cell culture model simulating immature teeth. Cytotoxic, migration, and angiogenic effects of Sodium hypochlorite associated with EDTA (NaOCl/EDTA), NaOCl associated with Smear Clear (NaOCl/SC), and QMix were compared. DESIGN: Three roots of mandibular first premolars had their length and root canal diameter standardized. Root canals were irrigated, and the resulting solutions were diluted in culture medium. Sulforhodamine B (SRB) assay was performed with apical papilla cells and with endothelial cells (HUVECs) to assess cytotoxicity. Polarity index and migration assays of apical papilla cells and sprouting of HUVECs were evaluated. Data were analyzed by ANOVA and Tukey post-hoc tests (p < .05). RESULTS: In apical papilla cells, NaOCl/SC and QMix promoted higher cytotoxicity, decreased fraction of elongated cells, and had lower migration speed and shorter migration distance of cells compared to NaOCl/EDTA. Also, HUVECs treated with NaOCl/SC and QMix showed decreased tubule formation in comparison with NaOCl/EDTA. CONCLUSIONS: NaOCl/SC and QMix showed unfavorable biological responses of cells involved in revascularization in comparison to NaOCl/EDTA. Further studies with other intracanal irrigants should be performed to improve the balance of root canal disinfection with biological responses.

Correction to: Periodontal Disease Impairs Muscle Recovery by Modulating the Recruitment of Leukocytes.

The original version of this article contained mistakes, and the authors would like to correct them.

Periodontal Disease Impairs Muscle Recovery by Modulating the Recruitment of Leukocytes.

The purpose of this study is to analyze the impact of periodontal disease (PD) associated with physical exercise on inflammatory mediators and muscle repair. Twenty-four Wistar rats were divided into four groups: control (SH), healthy trained (TH), sedentary with PD (SP), and trained with PD (TP). PD was induced in groups SP and TP while the trained groups performed treadmill exercises for 8 weeks. For the analysis of IL-6, IL-10, TNF-α, and leukocyte count, we collected blood samples. Cryolesions were induced in the tibialis anterior and gastrocnemius, which were analyzed for morphological changes. The presence of PD modified leukocyte counts, while exercise showed an additive role. PD increased levels of IL-6, IL-10, and TNF-α, and physical exercise changed only values of IL-10. The association between physical exercise and PD was responsible for an increased concentration of leukocytes in the region of the inflammation. Serum levels of inflammatory markers were modified by PD and, when combined with exercise, may negatively modulate inflammation. The association between PD and physical exercise showed the most significant changes in the number of inflammatory cells and may negatively influence the process of muscle repair.

NADPH-oxidase-derived ROS alters cell migration by modulating adhesions dynamics.

BACKGROUND INFORMATION: Cell migration requires the coordinated activation of structural and signalling molecules, such as the RhoGTPase Rac1. It is known that the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex assembly, which generates reactive oxygen species (ROS) at the cell membrane, also relies on Rac1 activation, indicating a possible effect of ROS during cell migration. In this study, we evaluated the effect of NADPH-oxidase-derived ROS on the migration process. RESULTS: Using time-lapse videos of CHO.K1 cells plated on fibronectin (2 µg/ml) or collagen (5 µg/cm2 ), we observed that depletion of ROS by N-acetyl-cysteine (NAC, 10 mM), an unspecific antioxidant, or diphenyliodonium (DPI, 10 µM), a NADPH-oxidase inhibitor, induced a ∼50% decrease in migration speed and severely impacted migration directionality. Then, we analysed the effects of NADPH oxidase on three migratory events: protrusion rate, adhesion process and signalling pathways related to cell migration. DPI induced an increase of ∼3 protrusion/cell, which were 2× faster but had a ∼50% retraction when compared with control. By pull-down assay, we observed no changes on Rac1 activation, indicating that ROS-mediated effects were related to downstream molecules, such as adhesion-related molecules. A reduction of the adhesion marker FAK-Y397 levels in cells treated with NAC and DPI was observed. In order to analyse adhesion dynamics, CHO.K1 cells transfected with paxillin-GFP analysed with total internal reflectance fluorescence (TIRF) indicated that DPI (5 µM) induced larger adhesions when compared with control. CONCLUSION: These results indicate that the local generation of NADPH-oxidase-derived ROS can modulate cell migration due to changes on adhesion dynamics and signalling. SIGNIFICANCE: This study highlights the physiological requirement of ROS for cell migration and the potential use of these molecules as targets to modulate the cell migration process at different diseases.

Low Doses of Curcuma longa Modulates Cell Migration and Cell-Cell Adhesion.

Cell invasion and metastasis are involved in clinical failures in cancer treatment, and both events require the acquisition of a migratory behavior by tumor cells. Curcumin is a promising natural product with anti-proliferative activity, but its effects on cell migration are still unclear. We evaluated the effects of curcumin on the proliferation, apoptosis, migration, and cell-cell adhesion of keratinocyte, oral squamous cell carcinoma (OSCC), and fibroblast cell lines, as well as in a xenograft model of OSCC. Curcumin (2 µM) decreased cell proliferation in cell lines with mesenchymal characteristics, while cell death was detected only at 50 µM. We observed that highly migratory cells showed a decrease on migration speed and directionality when treated with 2 or 5 µM of curcumin (50% and 40%, respectively, p < 0.05). Using spheroids, we observed that curcumin dose dependently decreased cell-cell adhesion, especially on tumor-derived spheroids. Also, in a xenograft model with patient-derived OSCC cells, the administration of curcumin decreased tumor growth and aggressiveness when compared with untreated tumors, indicating the potential antitumor effect in oral cancer. These results suggest that lower doses of curcumin can influence several steps involved in tumorigenesis, including migration properties, suggesting a possible use in cancer therapy. Copyright © 2017 John Wiley & Sons, Ltd.