Inverted Light Microscope Research Articles

Protective effect of Schistosoma mansoni infection and/or soluble egg antigen on allergic reaction in male mice

BackgroundInnovative treatments are being examined to develop more effective and innocuous protective medications for allergic conditions. In recent times, helminth-based immunotherapy is gaining attention as a potential therapeutic approach that could establish a pathway for controlling anaphylaxis. To the extent of our knowledge, there are no previous studies that examine the protective effect of both Schistosoma mansoni (S. mansoni) and its soluble egg antigen (SEA) together against anaphylaxis. Therefore, the purpose of the current study was to examine and compare the impact of SEA immunization and/or S. mansoni infection on Ovalbumin (OVA)-induced systemic anaphylaxes in mice model.ResultsThe outcome results revealed that S. mansoni infection and SEA immunization were able to improve body weight, reduce the mortality rate, increase plasma IgE and IgG4 levels and decrease histamine levels in broncho-alveolar lavage fluid (BALF). Additionally, they elevated interleukin-(IL)-4, IL-10, interferon-gamma (IFN-γ) and transforming growth factor-beta (ΤGF-β) levels in BALF. They also restored the stabilization of peritoneal mast cells (MCs) membrane in inverted light microscopy results accompanied by amelioration of the lung and liver histology.ConclusionsThe present study provided indication for the prophylactic effects of S. mansoni infection and SEA immunization against OVA-induced systemic anaphylaxes in mice model. Also, it focuses on the possible therapeutic mechanisms of helminth-derived products administration that might be related to upregulation of immune regulatory mechanisms. As a result, S. mansoni-derived products may be used as preventative supplemented treatments to inhibit the development of anaphylaxis which provides us with a new vision for developing pioneering therapy.

Biocompatibility of 3D-printed vs. thermoformed and heat-cured intraoral appliances

ObjectivesThe development of additive manufacturing has the potential to revolutionize the fabrication of medical devices. This technology, also known as 3D printing, offers precise, cost-effective, and personalized approaches, which could be particularly beneficial in the production of intraoral appliances. Despite its promise, research on the biocompatibility of 3D-printed intraoral devices is still limited. Our study aims to address this gap.MethodsWe examined the cytotoxicity of materials processed via three techniques commonly used for the fabrication of different intraoral appliances: 3D printing (Dental LT Clear), thermoforming (Duran adjusted with Durasplint LC), and conventional heat-curing (Villacryl H Plus). We also investigated the impact of chemical or UVC disinfection on the biocompatibility of these materials. We assessed the biological effects induced in human gingival fibroblasts (HGFs) through both direct contact tests (MTT and LDH assays) and extract tests (PrestoBlue, DCF, and cell death type assays). Additionally, we observed changes in cellular morphology and migration rate under an inverted light microscope. The surface roughness of materials was evaluated using contact profilometry. Statistical analysis was conducted using two-way analysis of variance.ResultsOur findings suggest that all three fabrication techniques induced a slight cytotoxic effect in HGFs, as evidenced by both direct contact and extract tests. However, these materials could be considered nontoxic according to the ISO 10993-5:2009 norm, as the decrease in metabolic activity observed was always less than 30% compared to the untreated control.ConclusionThis novel study confirms that 3D printing may be a safe alternative to conventional methods for fabricating intraoral appliances. However, further tests assessing the long-term intraoral usage are still needed.

Camel Whey Protein and Baicalein Suppressed Mast Cell Degranulation in Mice Models of IgE- and Non-IgE-Mediated Anaphylaxes: Potential Mechanisms on Downstream Cell Signaling of Mast Cells

ABSTRACT Introduction Novel treatments are being researched to develop more safe and effective protective medications for anaphylaxis. Camel whey protein (CWP) and baicalein (BAC, one of the major flavones) have multiple beneficial properties including anti-inflammatory and antioxidant activities. Methods The current study investigated/compared the therapeutic protection of repeated intragastric administration of CWP (100 mg/kg body weight, as an animal extract) and BAC (10 mg/kg body weight, as a plant extract), before the challenge with ovalbumin (OVA) or receiving the compound 48/80 (C48/80), against mice models for IgE-independent and dependent anaphylaxes. Besides, their effects on mast cells (MCs) downstream cell signaling were explored. Results The results revealed that CWP and BAC reduced the mortality rate, as compared with a MCs stabilizer “sulfasalazine (SSZ, 100 mg/kg body weight, intraperitoneally),” in both mice models. Furthermore, they prevented the MCs degranulation and significantly reduced (p < .05) lung tissue levels of cell signaling (p-AKT, p-ERK, and p-IκBα). Additionally, they decreased histamine, tryptase, leukotriene C4, prostaglandin D2, interleukin (IL)-4, and IL-10 levels in broncho-alveolar and peritoneal lavages in systemic anaphylaxis mice models. They also restored the stabilization of peritoneal MCs membrane in inverted light microscopy results accompanied by amelioration of the lung histology. Discussion The present study provided evidence for the protective therapeutic effect of CWP and BAC against anaphylaxis. As a result, CWP and BAC may be used as preventative supplemented regimens for both non-vegetarian and vegetarian consumers to treat allergy through downregulation of MCs signal transduction pathways, and hence controlling the production of inflammatory mediators.

Evaluation of Cardiospermum halicacabum on Bone Morphogenetic Protein-2 (BMP2) mRNA Expression in Osteoblast Cells.

Introduction Maintaining bone health is crucial for overall well-being, with osteoblasts playing a vital role in bone formation. Bone morphogenetic protein-2 (BMP2) is a key regulator, stimulating bone matrix synthesis and osteoblast differentiation. Recognizing BMP2's significance, there's growing interest in natural compounds, such as Cardiospermum halicacabum. This study explores Cardiospermum halicacabum's potential influence on BMP2 mRNA expression in osteoblast cells for insights into bone health modulation. Materials and methods This research utilized Cardiospermum halicacabum to explore its impact on MG-63 cells, a human osteoblast cell line. Osteoblast cells were cultured in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% heat-inactivated fetal bovine serum, and maintained at 37°C in a 5% CO2 and 95% air environment. Cell viability was evaluated by seeding osteoblast cells into 96-well plates and exposing them to different concentrations of Cardiospermum halicacabum (2.0 μg/ml and 20 μg/ml). The study observed both the promotion of osteoblast cell growth in MG-63 and morphological changes in thecells under an inverted light microscope at 10x magnification. Results were presented using one-way analysis of variance (ANOVA) conducted with IBM SPSS Statistics for Windows, Version 23 (Released 2015; IBM Corp., Armonk, New York, United States). Result The reverse transcription-polymerase chain (RT-PCR) results revealed an increased expression of BMP-2 mRNA fold change in comparison to the control group. A clear positive correlation was observed between the BMP-2 mRNA fold change and the notable increase in the concentration of Cardiospermum halicacabum. This investigation revealed a direct association of BMP-2 mRNA expressionwith the proliferation of osteoblast cells. Specifically, the BMP-2 mRNA fold change was recorded at 2.26±1.05 in Cardiospermum halicacabum at 2.0 μg/ml and 2.0 ± 0.84 at 20 μg/ml, with corresponding significances of 0.00, respectively. Conclusion Potential effects of Cardiospermum halicacabum on BMP-2 mRNA expression in osteoblast cellsand its role in bone health modulation revealed that Cardiospermum halicacabum may upregulate BMP-2 mRNA expression, suggesting its potential as a natural compound for enhancing bone formation. The observed positive correlation between Cardiospermum halicacabum concentration and BMP-2 mRNA fold change showed the significance of this botanical agent in promoting osteoblast cell proliferation. These results highlight the importance of further research to explore the applications of Cardiospermum halicacabum in managing bone disorders and improving overall bone health.

Biological Effects of PMMA and Composite Resins on Human Gingival Fibroblasts: An In Vitro Comparative Study.

The use of temporary resin for provisional restorations is a fundamental step to maintain the position of prepared teeth, to protect the pulpal vitality and the periodontal health as well as the occlusion. The present study aimed at evaluating the biological effects of two resins used in dentistry for temporary restorations, Coldpac (Yates Motloid) and ProTemp 4™ (3M ESPE ™), and their eluates, in an in vitro model of human gingival fibroblasts (hGFs). The activation of the inflammatory pathway NFκB p65/NLRP3/IL-1β induced by the self-curing resin disks was evaluated by real-time PCR, Western blotting and immunofluorescence analysis. The hGFs adhesion on resin disks was investigated by means of inverted light microscopy and scanning electron microscopy (SEM). Our results suggest that hGF cells cultured in adhesion and with eluate derived from ProTemp 4™ (3M ESPE ™) resin evidenced a downregulation in the expression of the inflammatory mediators such as NFκB p65, NLRP3 and IL-1β compared to the cells cultured with Coldpac (Yates Motloid) after 24 h and 1 week of culture. Furthermore, the cells cultured with ProTemp 4™ (3M ESPE ™) after 24 h and 1 week of culture reported a higher cell viability compared to the cells cultured with Coldpac (Yates Motloid), established by MTS cell analysis. Similar results were obtained when hGFs were placed in culture with the eluate derived from ProTemp 4™ (3M ESPE ™) resin which showed a higher cell viability compared to the cells cultured with eluate derived from Coldpac (Yates Motloid). These results highlighted the lower pro-inflammatory action and improved cell biocompatibility of ProTemp 4™ (3M ESPE ™), suggesting a better performance in terms of cells-material interaction.

Effectiveness of Chitosan versus Natural Aloe Vera on Candida Adherence in Denture Soft Lining Material.

Soft denture lining materials act as a cushion between the denture base and tissues. Alongside having many advantages, its main problem is candida growth due to its rubbery and porous texture. Many interventions have been performed to halt the growth of candida within soft lining materials such as the use of antifungal therapy and strict oral and denture hygiene but there are consequences such as recurrence, drug resistance, and toxicity related to these interventions. Since natural agents such as aloe vera and chitosan have been proven to have antibacterial and antifungal properties with minimum adverse effects, this study aimed to study the effectiveness of chitosan and aloe vera powders incorporated within denture soft lining materials against candida adherence. Methodology. A total of 60 soft-lining material samples were prepared that were equally divided into three groups, viz., group 1 (chitosan incorporation), group 2 (aloe vera incorporation), and group 3 (control). Candida was obtained from the microbiology lab to form a candidal suspension, diluted in 0.9% NaCl to match the McFarland standard bacteriologic solution. Samples were incubated at 37°C for 24 hours in test tubes containing 100 mL of the candidal suspension and 9.9 mL of the previously prepared Sabouraud dextrose agar. Crystal violet stain was used to stain the adhering cells by fixing them with methanol 80%. For each sample, the adhering candida cells were counted on three standard fields by using an inverted light microscope, and the mean of those fields was recorded. The mean value for samples containing aloe vera was 41.15, while the mean values for samples containing chitosan and the control group were 16.05 and 79.1, respectively. Of all the three groups, aloe vera powder had a significant efficacy against candida growth as compared to the chitosan and control groups (P value = 0.001). Both herbal agents were effective against candida growth. In comparison, aloe vera was more effective against candida growth compared to chitosan.

The influence of the environment on the diversity of Euglena species and their abundance in the lagoon of Nador-Morocco

The main objective of this work is to study the spatial and temporal evolution and distribution of Euglena species in the Nador lagoon. The study is based on four sampling stations and covers two specific seasons, namely spring and summer 2018. Euglenes belonging to the genus Euglena are particularly interesting and well known in the field of research, due to their great diversity in terms of pigmentation, size and morphological characteristics. Samples taken at each of the four sampling stations were carefully identified morphologically using an inverted light microscope. In total, five species belonging to the genus Euglena were identified, namely: Euglena viridis O.F. Müller 1786, Euglena Caudata Hübner 1886, Euglena Proxima P.A. Dangeard 1902, Euglena tuberculata Svirenko 1915, Euglena sp. Quantitative analysis of the species collected reveals some interesting results. The maximum cell density was recorded at station 4, located near the wastewater treatment plant, with a value of 55 cells per litre during the summer of 2019. In contrast, the minimum cell density was recorded at the same station 4, corresponding to Kariat Arekmen, with a value of 5 cells per litre during the spring period of 2019. These observations highlight significant variations in Euglenes cell density depending on geographical location and season.

Inhibition of lipopolysaccharide-induced NF-κB maintains osteogenesis of dental pulp and periodontal ligament stem cells.

Dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) can differentiate into osteoblasts, indicating that both are potential candidates for bone tissue engineering. Osteogenesis is influenced by many environmental factors, one of which is lipopolysaccharide (LPS). LPS-induced NF-κB activity affects the osteogenic potencies of different types of MSCs differently. This study evaluated the effect of LPS-induced NF-κB activity and its inhibition in DPSCs and PDLSCs. DPSCs and PDLSCs were cultured in an osteogenic medium, pretreated with/without NF-κB inhibitor Bay 11-7082, and treated with/without LPS. Alizarin red staining was performed to assess bone nodule formation, which was observed under an inverted light microscope. NF-κB and alkaline phosphatase (ALP) activities were measured to examine the effect of Bay 11-7082 pretreatment and LPS supplementation on osteogenic differentiation of DPSCs and PDLSCs. LPS significantly induced NF-κB activity (p = 0.000) and reduced ALP activity (p = 0.000), which inhibited bone nodule formation in DPSCs and PDLSCs. Bay 11-7082 inhibited LPS-induced NF-κB activity, and partially maintained ALP activity and osteogenic potency of LPS-supplemented DPSCs and PDLSCs. Thus, inhibition of LPS-induced NF-κB activity can maintain the osteogenic potency of DPSCs and PDLSCs.

SiRNA-Mediated B7H7 Knockdown in Gastric Cancer Lysate-Loaded Dendritic Cells Amplifies Expansion and Cytokine Secretion of Autologous T Cells.

Gastric cancer, ranked as the fifth most common cancer worldwide, presents multiple treatment challenges. These obstacles often arise due to cancer stem cells, which are associated with recurrence, metastasis, and drug resistance. While dendritic cell (DC)-based immunotherapy has shown promise as a therapeutic strategy, its efficacy can be limited by the tumor microenvironment and certain inhibitory immune checkpoint molecules, such as B7H7. SiRNA-medicated knockdown of B7H7 in tumor cell lysate-pulsed DCs can increase cytokine secretion and autologous T lymphocyte expansion. This study aimed to evaluate the impact of B7H7 suppression in gastric cancer cell lysate-pulsed DCs on the stimulatory potential of autologous CD3+ T lymphocytes. Peripheral blood mononuclear cells (PBMCs) were isolated and monocytes were obtained; then, they were differentiated to immature DCs (iDCs) by GM-CSF and IL-4. Tumor cell lysates from human gastric cancer cell lines were harvested, and iDCs were transformed into mature DCs (mDCs) by stimulating iDCs with tumor cell lysate and lipopolysaccharide. B7H7-siRNA was delivered into mDCs using electroporation, and gene silencing efficiency was assessed. The phenotypic characteristics of iDCs, mDCs, and B7H7-silenced mDCs were evaluated using specific surface markers, an inverted light microscope, and flow cytometry. CD3+ T cells were isolated via magnetically activated cell sorting. They were labeled with CFSE dye and co-cultured with mDCs and B7H7-silenced mDCs to evaluate their ability to induce T-cell proliferation. T-cell proliferation was assessed using flow cytometry. The concentration of TGF-β, IL-4, and IFN-γ secreted from CD3+ T cells in the co-cultured supernatant was evaluated to investigate the cytokine secretory activity of the cells. Transfection of B7H7 siRNA into mDCs was performed in optimal conditions, and the siRNA transfection effectively reduced B7H7 mRNA expression in a dose-dependent manner. SiRNA-mediated B7H7 knockdown in mDCs enhanced maturation and activation of the DCs, as demonstrated by an increased surface expression of CD11c, CD86, and CD40. Co-culture experiments revealed that B7H7-silenced mDCs had more capacity to induce T cell proliferation compared to non-transfected mDCs. The cytokine production patterns of T cells were also altered. Upon examining the levels of TGF-β, IL-4, and IFN-γ released by CD3+ T cells in the co-culture supernatant, we found that silencing B7H7 in mDCs resulted in a rise in IL-4 secretion and a reduction in TGF-β levels compared to mDCs that were not transfected. The study found that suppressing B7H7 expression in DCs significantly enhances their maturation and stimulatory activity when exposed to gastric cancer cell lysate. These B7H7-silenced DCs can substantially increase cytokine production and promote co-cultured T-cell expansion. Consequently, inhibiting B7H7 in DCs may offer a practical strategy to enhance the ability of DCs to initiate T lymphocyte responses and improve the effectiveness of DC-based cell therapy for cancer patients.

Evaluation the toxic effect of copper ions on the condition indices of benthic diatom Actinocyclus subtilis (W.Gregory) Ralfs 1861 in the experiment

Introduction. Pollution of marine coastal areas lead to the relevance of environmental monitoring including application of biotesting methods based on- the cultures of unicellular algae. Microalgae have different species-specific resistance to pollutants that expands application of different species as bioindicators of marine pollution.&#x0D; The aim of the study was to determine the threshold concentration of copper ions (Cu2+) for the survival and increase in the cells number of benthic diatom Actinocyclus subtilis (W.Gregory) Ralfs 1861 (Bacillariophyta) under the wide range of toxicant concentrations during 10-day toxicological experiments.&#x0D; Material and methods. The response of strain culture of the benthic diatom A. subtilis to various concentrations of copper sulfate (ranged from 16 to 1024 μg/l in terms of Cu2+ ions) was studied. In accordance with the previously developed protocol, the following indices were evaluated: alterations in the absolute number and proportion (%) of alive cells in the test-culture, as well as the specific growth rate in the number of A. subtilis cells at different concentrations of toxicant. Counting of alive and dead cells was carried out by micrographs taken for 12–15 random viewing fields under Nikon Eclipse inverted light microscope.&#x0D; Results. It was found that in the control and at concentration of copper ions 16 μg/l, the increase in the absolute number of cells in culture is described by sigmoid response curve. At the control еhe exponential growth phase occurs on days 5–7 and at concentration of 16 µg/l on days 3–5 of the experiment. The threshold concentration of copper ions (32 μg/l) which is critical for the survival of A. subtilis was determined, which is 3–7 times lower than threshold level for other benthic diatom species. At concentration of 32 µg/l, the phases of acceleration and exponential growth on the abundance curve are absent. The proportion of living cells in the culture decreases to 80% of the control level on day 3 and to 39% by day 10. At Cu2+ concentrations of 64 µg/l and above, sharp inhibition and death of culture is observed as early as 1–3 days. A positive specific growth rate of A. subtilis culture was revealed in the period of 1–5 days at copper concentration of 16 and &#x0D; 32 µg/l, and at concentration of 64 µg/l and higher the culture dies off. Negative values of the specific growth rate for all concentrations of the toxicant within the period of 5–10 days were obtained.&#x0D; Limitations. By the results of 10-day experiments the effect of 8 concentrations of copper sulfate on the culture of marine benthic diatom A. subtilis was studied. Three replicates in each concentration and exposure time were measured (1350 measurements in total), which is sufficient sampling for statistically reliable determination of the threshold values of copper ion toxicity for given test object.&#x0D; Conclusion. Considering the results obtained, the benthic diatom A. subtilis is highly sensitive to copper ions impact and can be recommended as new test-object for toxicology, as well as for application in monitoring of marine water areas subject to technogenic pollution.

Tangeretin's Anti-apoptotic Signaling Mechanisms in Oral Cancer Cells: In Vitro Anti-cancer Activity.

Introduction Citrus fruit peels contain Tangeretin, a natural chemical flavonoid that reinforces plant cell walls and serves as a defense mechanism. Apoptosis, growth inhibition, anti-oxidant, anti-diabetic, and anti-cancer activities are only a few of its many qualities. Tangeretin's principal function is to shield healthy cells or tissues from the harmful effects of chemotherapy. The purpose of this study was to investigate the apoptotic activity of Tangeretin's impact on KB (oral cancer cells) cell lines. Materials and method This study employed Tangeritin, in investigating its effects on oral cancer cells. Oral cancer cells were cultured in Dulbecco's modified Eagle's medium (DMEM), with 10% fetal bovine serum at 37°C in a 5% CO2 environment. Cell viability was assessed by seeding oral cancer cells in 96-well plates, exposing them to varying Tangeritin concentrations (50 µM, 100 µM, and 200 µM) with growth inhibition of KB cell viability in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assayand morphological changes in cells were observed under an inverted light microscope at 10x magnification. The results were reported as mean±standard error mean (SEM) using one-way analysis of variancethrough IBM SPSS Statistics for Windows, Version 23 (Released 2015; IBM Corp., Armonk, New York, United States). Result MTT assay showed a significant reduction in KB cell viability when treated with Tangeretin. With asignificant decrease in mRNA levels of the anti-apoptotic genes Bcl-2 and Bcl-xL. At 50 µM, 100 µM, and 200 µM, the levels of Bcl-2 were 0.85 ± 0.09, 0.62 ± 0.05, and 0.67 ± 0.05, respectively. Similarly, the mRNA expression of Bcl-xL was 0.82 ± 0.07 for 50 µM, 0.7 ± 0.06 for 100 µM, and 0.77 ± 0.06for 200 µM. The mRNA expression levels of Bax were 1.1 ± 0.09 for 50 µM, 1.4 ± 0.12for 100 µM, and 1.3 ± 0.11 for 200 µM, respectively. Conclusion Tangeretin showed a promising apoptotic activity in KB cells suggesting its utility as an anti-cancer compound. It prevented the growth and proliferation of cancer cells by acting on pro-apoptotic and anti-apoptotic genes. However, this conclusion is mostly based on the in vitro study. Therefore in vivo animal studies were needed to confirm the findings.

The influence of lemongrass essential oil addition into heat cured acrylic resin against Candida albicans adhesion

Background: For decades, the use of naturally accessible materials in treating human disease has been widespread. The goal of this study was to determine the anti-fungal effectiveness /of the lemongrass essential oil (LGEO) versus Candida albicans (C. albicans) adhesion to polymethylmethacrylate (PMMA) materials. Material and methods: LGEO's anti-fungal activity was tested against C. albicans adhesion using the following concentration of LGEO in PMMA monomer (2.5 vol. %, 5 vol. % LGEO) selected from the pilot study as the best two effective concentrations. A total of 40 specimens were fabricated for the candida adherence test and were subdivided into four equal groups: negative control 0 vol. % addition, experimental with 2.5 vol. % and 5 vol. % of LGEO addition and positive control with 1.4 wt. % nystatin addition. The sterile PMMA specimens were incubated at room temperature for 1 hr in sterile tubes with a sabouraud dextrose broth (SDA) in which a small amount of the yeast was isolated and suspended; under the inverted light microscope, the examination was done. The data were evaluated using a one-way ANOVA test, which showed a significant result at p&lt; 0.05. Results: The findings of the C. albicans adherence test exposed a considerable reduction in the number of C. albicans cells adhering to PMMA after adding 2.5 vol. % and 5 vol. % LGEO compared to specimens from the negative control and positive control groups at p&lt; 0.05. Conclusion: Adding LGEO into a heat-cure acrylic material can result in a denture base material with anti-fungal properties versus C. albicans microorganisms. The experimental group 5 vol. % LGEO additive showed the best anti-fungal activity

Green tea extract and hydroxyl-chloroquine combination enhances apoptosis in A549 non-small cell lung cancer cells.

Polyphenols, including catechins from green tea extract, have long been known for their potential anti-tumour activities. However, the precise mechanisms underlying their actions remain unclear. This study aimed to investigate the effects of green tea extract on A549 cells, a type of non-small lung cancer cells. A549 cells treated with green tea extract (GTE) were examined using an inverted light microscope and a fluorescence microscope. Cell sensitivity was evaluated using the MTT assay, while cell death was assessed using the Tali image-based cytometer. Ultra structural changes were observed using a transmission electron microscope. The findings suggested that even at the highest dose tested (150 µM), GTE did not exhibit toxic effects on A549 cells. Likewise, treatment with GTE resulted in a minimal, dose-dependent increase in the population of apoptotic cells. However, the analysis of cell structures using light and electron microscopy revealed an enhanced accumulation of vacuole-like structures in response to GTE. Moreover, under the fluorescence microscope, an increase in acidic vesicular organelles and the formation of LC3-II puncta were observed following GTE treatment. Assessment of autophagy function indicated that GTE-induced autophagy may serve as a self-protective mechanism against cytotoxicity, as blocking autophagy with bafilomycin A1 reduced cell viability and enhanced necrotic cell death in response to GTE treatment. In summary, our results demonstrate that A549 cells are insensitive to both low and high concentrations of green tea extract, likely due to the induction of cytoprotective autophagy. These findings suggest that the potential utility of GTE in lung cancer therapy may lie in its synergistic combinations with drugs or small molecules that target autophagy, rather than as a standalone therapy.

Polyhydroxylated Fullerene C60(OH)40 Nanofilms Promote the Mesenchymal-Epithelial Transition of Human Liver Cancer Cells via the TGF-β1/Smad Pathway.

The various growth factors change the phenotype of neoplastic cells from sedentary (epithelial) to invasive (mesenchymal), which weaken intercellular connections and promote chemotaxis. It can be assumed that the use of anti-inflammatory polyhydroxyfull nanofilms will restore the sedentary phenotype of neoplastic cells in the primary site of the tumor and, consequently, increase the effectiveness of the therapy. The studies were carried out on liver cancer cells HepG2, C3A and SNU-449, and non-cancer hepatic cell line THLE-3. Transforming growth factor (TGF), epidermal growth factor and tumor necrosis factor were used to induce the epithelial-mesenchymal transition. C60(OH)40 nanofilm was used to induce the mesenchymal-epithelial transition. Obtaining an invasive phenotype was confirmed on the basis of changes in the morphology using inverted light microscopy. RT-PCR was used to confirm mesenchymal or epithelial phenotype based on e-cadherin, snail, vimentin expression or others. Water colloids at a concentration of 100 mg/L were used to create nanofilms of fullerene, fullerenol, diamond and graphene oxide. The ELISA test for the determination of TGF expression and growth factor antibody array were used to select the most anti-inflammatory carbon nanofilm. Mitochondrial activity and proliferation of cells were measured by XTT and BrdU tests. Cells lost their natural morphology of cells growing in clusters and resembled fibroblast cells after adding a cocktail of factors. Among the four allotropic forms of carbon tested, only the C60(OH)40 nanofilm inhibited the secretion of TGF in all the cell lines used and inhibited the secretion of other factors, including insulin-like growth factor system. Nanofilm C60(OH)40 was non-toxic to liver cells and inhibited the TGF-β1/Smad pathway of invasive cells treated with the growth factor cocktail. The introduction of an anti-inflammatory, nontoxic component that can induce the mesenchymal-epithelial transition of cancer cells may represent a future adjuvant therapy after tumor resection.

Enriched Graphene Oxide-Polypropylene Suture Threads Buttons Modulate the Inflammatory Pathway Induced by Escherichia coli Lipopolysaccharide.

Graphene oxide (GO), derived from graphene, has remarkable chemical-physical properties such as stability, strength, and thermal or electric conductivity and additionally shows antibacterial and anti-inflammatory properties. The present study aimed to evaluate the anti-inflammatory effects of polypropylene suture threads buttons (PPSTBs), enriched with two different concentrations of GO, in the modulation of the inflammatory pathway TLR4/MyD 88/NFκB p65/NLRP3 induced by the Escherichia coli (E. coli) lipopolysaccharide (LPS-E). The gene and the protein expression of inflammatory markers were evaluated in an in vitro model of primary human gingival fibroblasts (hGFs) by real-time PCR, western blotting, and immunofluorescence analysis. Both GO concentrations used in the polypropylene suture threads buttons-GO constructs (PPSTBs-GO) decreased the expression of inflammatory markers in hGFs treated with LPS-E. The hGFs morphology and adhesion on the PPSTBs-GO constructs were also visualized by inverted light microscopy, scanning electron microscopy (SEM), and real-time PCR. Together, these results suggest that enriched PPSTBs-GO modulates the inflammatory process through TLR4/MyD 88/NFκB p65/NLRP3 pathway.

Synergistic effect of polymer functionalized graphene oxide system for breast cancer treatment

The multifaceted drug carrier system is an emerging trend in delivering chemotherapeutic drugs and photosensitizers for the synergistic effect. In this work, we have designed a functionalized graphene oxide (GO) based carrier system for combined chemo-photodynamic therapeutic effects. Doxorubicin (DOX) and rose bengal (RB) were entrapped on the surface of GO via hydrophobic and π-π stacking interactions. The functional group determination, crystalline properties, surface morphology, and hydrodynamic size were evaluated using FT-IR, XRD, SEM, TEM, AFM, and DLS analysis. At 24 h, the entrapment efficiency was 65 % DOX and 40.92 % RB, and the loading capacities were 16.9 % DOX and 5.68 % RB observed at 30 min. The drug release percentage was higher in pH-2.6 rather than in pH-5.5, 6.8, and 7.4 pH environments. The in-vitro toxicity analysis using the LDH assay reveals that the DOX and RB co-loaded carriers had a significant cytotoxic effect on MCF-7 cells, indicating that the carrier could improve the therapeutic efficacy of DOX. Morphological changes were studied using inverted light microscopy; the cells were irradiated with a laser 525 nm 10 J/cm2 for 2 min 51 sec, and it was observed that the DOX and RB co-loaded carrier with laser-irradiated cells exposed the high-level morphological changes with the occurrence of apoptotic cell death. Compared to free DOX, the DOX/RB co-loaded carrier + laser had an efficient anticancer activity, as confirmed by DAPI staining cell uptake, flow cytometry, and intracellular ROS generation analysis. The DOX and RB co-loaded carrier clearly exhibits the RB-mediated photodynamic action on MCF-7 cells in response to external laser light irradiation. It permits an on-demand dual-payload release to trigger an instantaneous photodynamic and chemo treatment for cancer cell eradication. Finally, the ensuing dual-agent release is probable to successfully fight cancer via a synergistic effect.

FAM83G-based Peptide Induces Apoptosis on Cultured Liver Cancer Cell.

Previously, AF-956, which contains S356 of FAM83G and an N-terminal antenna peptide for entry into colon cancer cells, is markedly antiproliferative compared to a control peptide (AF-859), which lacks the N-terminal antenna peptide, by inducing apoptosis via the inhibition of HSP27 phosphorylation at residues S15 and S82. Because FAM83G-derived peptides are promising lead compounds for colon cancer treatment, we reanalyzed the effect of AG-066, which contains S356 of FAM83G and an N-terminal antenna peptide for entry into the liver cancer cells. HepG2 liver cancer cells were incubated with either AF-859 or AG-066 at a concentration of 54 μM at 37 °C for 24, 48, and 72 h. The effects of AF-859 and AG-066 on the cultured HepG2 cells were estimated using an inverted light microscope. Furthermore, the DNA ladder method and the dead cell assay were performed by applying Live/Dead Cell Staining Kit II. Erk phosphorylation was estimated by western blotting. Treatment with AG-066 markedly reduced HepG2 viable cell counts compared to the AF- 859-treated HepG2 cells, as evident from the significantly increased number of dead cells in the culture medium. Additionally, AG-066 treatment increased cellular DNA laddering. We found no difference in Erk phosphorylation status between the AG-066- and AF-859-treated groups. This study illustrated that the peptide with a structure based on FAM83G functions as a spontaneous apoptosis inducer for liver cancer cells. Hence, it is a promising lead compound for the treatment of liver cancer.

Photodynamic Therapy Efficacy of Novel Zinc Phthalocyanine Tetra Sodium 2-Mercaptoacetate Combined with Cannabidiol on Metastatic Melanoma.

This work reports for the first time on the synthesis, characterization, and photodynamic therapy effect of a novel water-soluble zinc (II) 2(3), 9(10), 16(17), 23(24)-tetrakis-(sodium 2-mercaptoacetate) phthalocyanine (ZnPcTS41), on metastatic melanoma cells (A375) combined with cannabidiol (CBD). The ZnPcTS41 structure was confirmed using FTIR, NMR, MS, and elemental analysis while the electronic absorption spectrum was studied using UV-VIS. The study reports further on the dose-dependent effects of ZnPcTS41 (1-8 µM) and CBD alone (0.3-1.1 µM) at 636 nm with 10 J/cm2 on cellular morphology and viability. The IC50 concentrations of ZnPcTS41 and CBD were found to be 5.3 µM and 0.63 µM, respectively. The cytotoxicity effects of the ZnPcTS41 enhanced with CBD on A375 cells were assessed using MTT cell viability assay, ATP cellular proliferation and inverted light microscopy. Cell death induction was also determined via Annexin V-FITC-PI. The combination of CBD- and ZnPcTS41-mediated PDT resulted in a significant reduction in cell viability (15%***) and an increase in the late apoptotic cell population (25%*). These findings suggest that enhancing PDT with anticancer agents such as CBD could possibly obliterate cancer cells and inhibit tumor recurrence.

Photodynamic Effects of Thuja occidentalis on Lung Cancer Cells.

The global incidence and mortality rates resulting from lung cancer encapsulate a need to identify more effective treatment protocols. Photodynamic therapy (PDT) and homeopathy offer possible anticancer therapies as part of a multi-disciplinary approach. Studies have identified the anticancer effects of Thuja occidentalis L. plant extracts. The aim of this study was to investigate the effects of Thuja occidentalis (TO) homeopathic mother tincture and TO mediated PDT (TO-PDT) on A549 lung cancer cells. Commercially available A549 cells were pre-treated with TO, or laser irradiation at 660 nm, or the combined treatment (TO-PDT). Cells were analyzed morphologically by inverted light microscopy and Hoechst stain; and biochemically by lactate dehydrogenase (LDH), adenosine triphosphate (ATP), and trypan blue assays. Cells treated with TO and TO-PDT demonstrated morphological changes in the cell and cell nuclei indicative of cell death. These groups exhibited a dose dependent increase in LDH release and a decrease in ATP levels and cell viability indicating its cytotoxic and antiproliferative potential. Furthermore, at the same doses, TO when photoactivated in PDT induced enhanced anticancer responses thereby surpassing the effects of treatment with the tincture alone. Results demonstrate how the direct cytotoxic effects of TO can be improved when administered as a photosensitizer in PDT to promote cancer cell death.

Protective Effect of Salvianolic Acid A against N-Methyl-N-Nitrosourea-Induced Retinal Degeneration.

Objective Retinal degeneration (RD) is a serious, irreversible, and blinding eye disease, which seriously affects the visual function and quality of life of patients. At present, there is no effective method to treat RD. The final outcome of its development is photoreceptor cell oxidation and apoptosis. Therefore, looking for safe, convenient, and effective antioxidant therapy is still the key research field of Rd. In this study, the mice model of RD was induced by N-methyl-N-nitrosourea (MNU) in vivo to explore the therapeutic effect and mechanism of salvianolic acids (Sal A) on RD. In vitro, the protective effect of Sal A on MNU injured 661 W cell line of mouse retina photoreceptor cone cells was investigated preliminarily. Methods Male C57BL/6 mice (7–8 weeks old) received a single intraperitoneal injection (ip) of 60 mg/kg MNU or vehicle control. Treatment groups then received Sal-A 0.5 mg/kg and 1.0 mg/kg via daily intravenous injections. On day 7, functional and morphological examinations were performed, including photopic and scotopic electroretinography (ERG) and hematological analyses to observe functional changes and damage to the outer nuclear layer (ONL). On the 3rd and 7th days, the levels of superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were determined. The expression of retinal Bax, Bcl-2, and caspase-3 was quantified by Western blot and RT-PCR assays. 661 W strain of mice retinal photoreceptor cone cells were cultured in vitro and treated with 1 µm MNU. The cells in the treatment group were given 50 μM Sal A as an intervention. The growth of 661 W cells was observed and recorded under an inverted light microscope, and the activity of cells was detected by the MTT method. Results Sal A treatment was effective against MNU-induced RD in mice at both 0.5 mg/kg/d and 1.0 mg/kg/d doses, and the protective effect was dose-dependent. Sal A can alleviate MNU-mediated alterations to retinal ERG activity and can support maintenance of the thickness of the ONL layer. Sal A treatment increases the expression of retinal SOD and reduces the lipid peroxidation product MDA, suggesting that its protective effect is related to the oxidation resistance. It can offset changes to the expression of apoptotic factors in the retina caused by MNU treatment. Sal A mitigates MNU-mediated damage to cultured mice photoreceptor cone cells 661 W in vitro. Conclusion Sal A alleviates the damage caused by MNU to retinal photoreceptor cells in vivo and in vivo, and its protective effect is related to its antioxidant and antiapoptotic activities.