Biocompatible graphene-zirconia nanocomposite as a cyto-safe immunosensor for the rapid detection of carcinoembryonic antigen

AbstractGraphene-based materials have gained remarkable attention in numerous disciplines owing to their unique electrochemical properties. Out of various hybridized nanocomposites, graphene-zirconia nanocomposite (GZ) was distinctive due to its biocompatibility. Zirconia nanoparticles serve as spacers that reduce the stacking of graphene and improve the electrochemical performance of the material. Considering that lungs and skin suffer the greatest exposure to nanoparticles, this study aimed to evaluate the cytotoxicity of the as-synthesized GZ nanocomposites on MRC5 (lung cells) and HaCaT (skin cells) via morphological observation and cell viability assay using 3-(4,5 dimethylthiazol-2-yl)-(2,5-diphenyltetrazolium bromide) tetrazolium (MTT). GZ-treated cells showed a comparable proliferation rate and morphology with untreated cells under microscopic evaluation. Based on MTT results, the IC50 values of GZ were > 500 µg/ml for MRC5 and HaCaT cells. The excellent biocompatibility was the supremacy of GZ over other nanocomposites applied as electrode materials in biosensors. GZ was functionalized with biolinker for the detection of carcinoembryonic antigen (CEA). The proposed immunosensor exhibited good responses towards CEA detection, with a 4.25 pg/ml LOD and correlation coefficient of R2 = 0.99 within a linear working range from 0.01 to 10 ng/ml. The performance of the immunosensor to detect CEA present in human serum was also evaluated. Good recovery of CEA was found, suggesting that the proposed immunosensor possess a high affinity to CEA even in a complex biological matrix, rendering it a promising sensing platform for real sample analysis and open a new way for the detection of cancer-associated proteins.

Download Full-text

Modulation of Hydrogen Peroxide-Induced Oxidative Stress in Human Neuronal Cells by Thymoquinone-Rich Fraction and Thymoquinone via Transcriptomic Regulation of Antioxidant and Apoptotic Signaling Genes

Oxidative Medicine and Cellular Longevity ◽

10.1155/2016/2528935 ◽

2016 ◽

Vol 2016 ◽

pp. 1-15 ◽

Cited By ~ 16

Author(s):

Norsharina Ismail ◽

Maznah Ismail ◽

Nur Hanisah Azmi ◽

Muhammad Firdaus Abu Bakar ◽

Hamidon Basri ◽

...

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Nigella Sativa ◽

Morphological Observation ◽

Cell Viability Assay ◽

Viability Assay ◽

Expression Of Genes ◽

Intracellular Ros ◽

Human Neuronal Cells ◽

Better Than

Nigella sativaLinn. (N. sativa) and its bioactive constituent Thymoquinone (TQ) have demonstrated numerous pharmacological attributes. In the present study, the neuroprotective properties of Thymoquinone-rich fraction (TQRF) and TQ against hydrogen peroxide- (H2O2-) induced neurotoxicity in differentiated human SH-SY5Y cells were investigated. TQRF was extracted using supercritical fluid extraction while TQ was acquired commercially, and their effects on H2O2were evaluated using cell viability assay, reactive oxygen species (ROS) assay, morphological observation, and multiplex gene expression. Both TQRF and TQ protected the cells against H2O2by preserving the mitochondrial metabolic enzymes, reducing intracellular ROS levels, preserving morphological architecture, and modulating the expression of genes related to antioxidants (SOD1, SOD2, and catalase) and signaling genes (p53, AKT1, ERK1/2, p38 MAPK, JNK, and NF-κβ). In conclusion, the enhanced efficacy of TQRF over TQ was likely due to the synergism of multiple constituents in TQRF. The efficacy of TQRF was better than that of TQ alone when equal concentrations of TQ in TQRF were compared. In addition, TQRF also showed comparable effects to TQ when the same concentrations were tested. These findings provide further support for the use of TQRF as an alternative to combat oxidative stress insults in neurodegenerative diseases.

Download Full-text

Proliferative Effect of Tilapia Fish (Oreochromis niloticus) Lectin on BALB/c Mice Splenocytes

Protein and Peptide Letters ◽

10.2174/0929866526666190911144057 ◽

2019 ◽

Vol 26 (12) ◽

pp. 887-892

Author(s):

Cynarha Daysy Cardoso da Silva ◽

Cristiane Moutinho Lagos de Melo ◽

Elba Verônica Matoso Maciel Carvalho ◽

Mércia Andréa Lino da Silva ◽

Rosiely Félix Bezerra ◽

...

Keyword(s):

Cell Proliferation ◽

Oreochromis Niloticus ◽

Cytokine Production ◽

Thymidine Incorporation ◽

Con A ◽

Cell Viability Assay ◽

Proliferative Effect ◽

Viability Assay ◽

Apoptosis And Necrosis

Background: Lectins have been studied in recent years due to their immunomodulatory activities. Objective: We purified a lectin named OniL from tilapia fish (Oreochromis niloticus) and here we analyzed the cell proliferation and cytokine production in Balb/c mice splenocytes. Methods: Cells were stimulated in vitro in 24, 48, 72 hours and 6 days with different concentrations of OniL and Con A. Evaluation of cell proliferation was performed through [3H]-thymidine incorporation, cytokines were investigated using ELISA assay and cell viability assay was performed by investigation of damage through signals of apoptosis and necrosis. Results: OniL did not promote significant cell death, induced high mitogenic activity in relation to control and Con A and stimulated the cells to release high IL-2 and IL-6 cytokines. Conclusion: These findings suggest that, like Con A, OniL lectin can be used as a mitogenic agent in immunostimulatory assays.

Download Full-text

Do We have a Satisfactory Cell Viability Assay? Review of the Currently Commercially-Available Assays

Current Drug Discovery Technologies ◽

10.2174/1570163815666180925095433 ◽

2020 ◽

Vol 17 (1) ◽

pp. 2-22 ◽

Cited By ~ 3

Author(s):

Abdel-Baset Halim

Keyword(s):

Cell Viability ◽

Data Interpretation ◽

Positive Control ◽

Live Cells ◽

Proof Of Concept ◽

Cell Viability Assay ◽

Viability Assay ◽

Current Cell ◽

Dying Cells ◽

Differential Permeability

:Cell-based assays are an important part of the drug discovery process and clinical research. One of the main hurdles is to design sufficiently robust assays with adequate signal to noise parameters while maintaining the inherent physiology of the cells and not interfering with the pharmacology of target being investigated.:A plethora of assays that assess cell viability (or cell heath in general) are commercially available and can be classified under different categories according to their concepts and principle of reactions. The assays are valuable tools, however, suffer from a large number of limitations. Some of these limitations can be procedural or operational, but others can be critical as those related to a poor concept or the lack of proof of concept of an assay, e.g. those relying on differential permeability of dyes in-and-out of viable versus compromised cell membranes. While the assays can differentiate between dead and live cells, most, if not all, of them can just assess the relative performance of cells rather than providing a clear distinction between healthy and dying cells. The possible impact of relatively high molecular weight dyes, used in most of the assay, on cell viability has not been addressed. More innovative assays are needed, and until better alternatives are developed, setup of current cell-based studies and data interpretation should be made with the limitations in mind. Negative and positive control should be considered whenever feasible. Also, researchers should use more than one orthogonal method for better assessment of cell health.

Download Full-text

SPARC regulates ferroptosis induced by sorafenib in human hepatocellular carcinoma

Cancer Biomarkers ◽

10.3233/cbm-200101 ◽

2021 ◽

pp. 1-9

Author(s):

Hong-Wei Hua ◽

Hao-Sheng Jiang ◽

Ling Jia ◽

Yi-Ping Jia ◽

Yu-Lan Yao ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Molecular Mechanism ◽

Cancer Progression ◽

Hepg2 Cells ◽

Cytotoxic Effect ◽

Cell Viability Assay ◽

Viability Assay ◽

Ldh Release ◽

Related Proteins ◽

Hcc Cells

BACKGROUND: Secreted protein acidic and rich in cysteine (SPARC) is implicated in cancer progression, but its role and associated molecular mechanism in the sorafenib sensitivity of hepatocellular carcinoma cells (HCC) remains elusive. METHODS: Human HCC cell lines Hep3B and HepG2 were treated with sorafenib alone or combined with activator or inhibitor of ferroptosis. Cell viability assay, reactive oxygen species (ROS) assay, lactate dehydrogenase (LDH) assay and western blot were used to study the regulatory mechanism of SPARC on HCC cells. RESULTS: Overexpression of SPARC enhanced the cytotoxic effect of sorafenib in Hep3B and HepG2 cells compared with parental cells. Depletion of SPARC decreased the cytotoxic effect of sorafenib in Hep3B and HepG2 cells compared with parental cells. Moreover, overexpression of SPARC significantly induced LDH release, whereas depletion of SPARC suppressed the release of LDH in Hep3B and HepG2 cells. Inhibition of ferroptosis exerted a clear inhibitory role against LDH release, whereas activation of ferroptosis promoted the release of LDH in HCC cells, as accompanied with deregulated expression of ferroptosis-related proteins. Furthermore, overexpression of SPARC induced oxidative stress, whereas depletion of SPARC suppressed the production of ROS. Deferoxamine (DFX)-induced inhibition of ferroptosis suppressed the production of ROS, while activation of ferroptosis promoted the contents of ROS in HCC cells exposed to sorafenib. CONCLUSION: Our findings give a better understanding of ferroptosis and its molecular mechanism in HCC cells that is regulated by SPARC in response to sorafenib.

Download Full-text

Differential Effects of Halofuginone Enantiomers on Muscle Fibrosis and Histopathology in Duchenne Muscular Dystrophy

International Journal of Molecular Sciences ◽

10.3390/ijms22137063 ◽

2021 ◽

Vol 22 (13) ◽

pp. 7063

Author(s):

Sharon Mordechay ◽

Shaun Smullen ◽

Paul Evans ◽

Olga Genin ◽

Mark Pines ◽

...

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Motor Coordination ◽

Mdx Mice ◽

Cell Viability Assay ◽

Muscle Fibrosis ◽

Antifibrotic Therapy ◽

Racemic Form ◽

Viability Assay

Progressive loss of muscle and muscle function is associated with significant fibrosis in Duchenne muscular dystrophy (DMD) patients. Halofuginone, an analog of febrifugine, prevents fibrosis in various animal models, including those of muscular dystrophies. Effects of (+)/(−)-halofuginone enantiomers on motor coordination and diaphragm histopathology in mdx mice, the mouse model for DMD, were examined. Four-week-old male mice were treated with racemic halofuginone, or its separate enantiomers, for 10 weeks. Controls were treated with saline. Racemic halofuginone-treated mice demonstrated better motor coordination and balance than controls. However, (+)-halofuginone surpassed the racemic form’s effect. No effect was observed for (−)-halofuginone, which behaved like the control. A significant reduction in collagen content and degenerative areas, and an increase in utrophin levels were observed in diaphragms of mice treated with racemic halofuginone. Again, (+)-halofuginone was more effective than the racemic form, whereas (−)-halofuginone had no effect. Both racemic and (+)-halofuginone increased diaphragm myofiber diameters, with no effect for (−)-halofuginone. No effects were observed for any of the compounds tested in an in-vitro cell viability assay. These results, demonstrating a differential effect of the halofuginone enantiomers and superiority of (+)-halofuginone, are of great importance for future use of (+)-halofuginone as a DMD antifibrotic therapy.

Download Full-text

BDMC protects AD in vitro via AMPK and SIRT1

Translational Neuroscience ◽

10.1515/tnsci-2020-0140 ◽

2020 ◽

Vol 11 (1) ◽

pp. 319-327

Author(s):

Chenlin Xu ◽

Zijian Xiao ◽

Heng Wu ◽

Guijuan Zhou ◽

Duanqun He ◽

...

Keyword(s):

Neurodegenerative Disorder ◽

Neuroprotective Effect ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Activity Measurement ◽

Therapeutic Approaches ◽

Cell Viability Assay ◽

Viability Assay ◽

Compound C

AbstractBackgroundAlzheimer’s disease (AD) is a common neurodegenerative disorder without any satisfactory therapeutic approaches. AD is mainly characterized by the deposition of β-amyloid protein (Aβ) and extensive neuronal cell death. Curcumin, with anti-oxidative stress (OS) and cell apoptosis properties, plays essential roles in AD. However, whether bisdemethoxycurcumin (BDMC), a derivative of curcumin, can exert a neuroprotective effect in AD remains to be elucidated.MethodsIn this study, SK-N-SH cells were used to establish an in vitro model to investigate the effects of BDMC on the Aβ1–42-induced neurotoxicity. SK-N-SH cells were pretreated with BDMC and with or without compound C and EX527 for 30 min after co-incubation with rotenone for 24 h. Subsequently, western blotting, cell viability assay and SOD and GSH activity measurement were performed.ResultsBDMC increased the cell survival, anti-OS ability, AMPK phosphorylation levels and SIRT1 in SK-N-SH cells treated with Aβ1–42. However, after treatment with compound C, an AMPK inhibitor, and EX527, an SIRT1inhibitor, the neuroprotective roles of BDMC on SK-N-SH cells treated with Aβ1–42 were inhibited.ConclusionThese results suggest that BDMC exerts a neuroprotective role on SK-N-SH cells in vitro via AMPK/SIRT1 signaling, laying the foundation for the application of BDMC in the treatment of neurodegenerative diseases related to AMPK/SIRT1 signaling.

Download Full-text

Long-Term Cytotoxicity, pH and Dissolution Rate of AH Plus and MTA Fillapex

Brazilian Dental Journal ◽

10.1590/0103-6440201600735 ◽

2016 ◽

Vol 27 (4) ◽

pp. 419-423 ◽

Cited By ~ 10

Author(s):

Emmanuel João Nogueira Leal da Silva ◽

Thais Accorsi-Mendonça ◽

Ana Carolina Pedrosa ◽

José Mauro Granjeiro ◽

Alexandre A. Zaia

Keyword(s):

Dissolution Rate ◽

Experimental Period ◽

Cell Viability Assay ◽

Time Points ◽

Viability Assay ◽

Significant Difference ◽

High Dissolution Rate ◽

Post Test ◽

Ah Plus

Abstract The aim of the present study was to verify the long-term cytotoxic effects of the MTA Fillapex and to compare them with AH Plus. Dissolution rate and pH were also evaluated. Human osteoblast cells were incubated with elutes of fresh specimens from AH Plus and MTA Fillapex, and with elutes of the same specimens for 4 successive weeks. Elute's pH was evaluated at each time point. A multiparametric cell viability assay was performed. For dissolution rate, ISO methodology was used. The results were analyzed by one-way analysis of variance, complemented with the Tukey post-test (p<0.05). No significant difference was found among the materials when fresh mixed (p>0.05). After 1 week, AH Plus became non-cytotoxic on all three evaluated parameters. Conversely, MTA Fillapex remained severely and mildly cytotoxic over the entire experimental period (p<0.05). The dissolution rate of AH Plus was significantly lower than MTA Fillapex at all time points (p>0.05). The pH of AH Plus was significantly lower than MTA Fillapex at the second and third week (p<0.05). In the other tested time points no statistical difference was observed. In conclusion, MTA Fillapex remained cytotoxic after 4 weeks and its cytotoxicity may be related to the high dissolution rate of this material.

Download Full-text

Human Recombinant Fab Fragment Neutralizes Shiga Toxin Type 2 Cytotoxic Effects in vitro and in vivo

Toxins ◽

10.3390/toxins10120508 ◽

2018 ◽

Vol 10 (12) ◽

pp. 508 ◽

Cited By ~ 4

Author(s):

Daniela Luz ◽

Maria Amaral ◽

Flavia Sacerdoti ◽

Alan Bernal ◽

Wagner Quintilio ◽

...

Keyword(s):

Shiga Toxin ◽

Lethal Dose ◽

Dependent Manner ◽

Fab Fragment ◽

Cell Viability Assay ◽

Cytotoxic Effects ◽

Morphological Alterations ◽

Viability Assay

Shiga toxin (Stx) producing Escherichia coli (STEC) is responsible for causing hemolytic uremic syndrome (HUS), a life-threatening thrombotic microangiopathy characterized by thrombocytopenia, hemolytic anemia, and acute renal failure after bacterially induced hemorrhagic diarrhea. Until now, there has been neither an effective treatment nor method of prevention for the deleterious effects caused by Stx intoxication. Antibodies are well recognized as affinity components of therapeutic drugs; thus, a previously obtained recombinant human FabC11:Stx2 fragment was used to neutralize Stx2 in vitro in a Vero cell viability assay. Herein, we demonstrated that this fragment neutralized, in a dose-dependent manner, the cytotoxic effects of Stx2 on human glomerular endothelial cells, on human proximal tubular epithelial cells, and prevented the morphological alterations induced by Stx2. FabC11:Stx2 protected mice from a lethal dose of Stx2 by toxin-antibody pre-incubation. Altogether, our results show the ability of a new encouraging molecule to prevent Stx-intoxication symptoms during STEC infection.

Download Full-text

Anti-inflammatory effects of Anredera cordifolia and Piper crocatum extracts on lipopolysaccharide-stimulated macrophage cell line

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v12i1.28714 ◽

2017 ◽

Vol 12 (1) ◽

pp. 35 ◽

Cited By ~ 7

Author(s):

Dian Ratih Laksmitawati ◽

Anisa Widyastuti ◽

Nadia Karami ◽

Ervi Afifah ◽

Dwi Davidson Rihibiha ◽

...

Keyword(s):

Cell Line ◽

Tumor Necrosis ◽

Macrophage Cell Line ◽

Macrophage Cell ◽

Cell Viability Assay ◽

Viability Assay ◽

Tnf Α ◽

Anti Inflammatory ◽

Α Level ◽

Necrosis Factor

In this study, the anti-inflammatory potential of Anredera cordifolia and Piper crocatum extracts on lipopolysaccharide-induced murine macrophage cell line (RAW 264.7) was observed. Cell viability assay was performed with MTS assay. Parameters measured to determine the anti-inflammatory activity were interleukin-1β (IL-1β), tumor necrosis factor (TNF)-α, nitric oxide (NO) and IL-6. Both A. cordifolia and P. crocatum at concentration of 50 µg/mL in cell line resulted significant decrease in TNF-α level (250.3 and 242.5 pg/mL respectively). A. cordifolia showed significant decrease in IL-1β level at 50 µg/mL and IL-6 level at 10 µg/mL, whilst P. crocatum showed significant decrease IL-1β level in three concentrations with lowest level at 50 µg/mL. A. cordifolia showed lowest decrease in NO level at 50 µg/mL but not comparable with normal cells, whilst P. crocatum showed significant decrease in NO level at 50 µg/mL. This research revealed that A. cordifolia and P. crocatum possess the anti-inflammatory potential indicated by the inhibitory activity of the inflammatory mediators including, TNF-α, IL-1β, IL-6, and NO.

Download Full-text

Promising biocompatible hydrogels of crosslinked polyelectrolytes for biomedical applications

Current Directions in Biomedical Engineering ◽

10.1515/cdbme-2017-0147 ◽

2017 ◽

Vol 3 (2) ◽

pp. 695-698

Author(s):

Andreas Brietzke ◽

Christian von der Ehe ◽

Sabine Illner ◽

Claudia Matschegewski ◽

Niels Grabow ◽

...

Keyword(s):

Aqueous Solution ◽

Ionic Liquids ◽

Cell Viability ◽

Biomedical Applications ◽

High Potential ◽

Cell Viability Assay ◽

Mouse Fibroblasts ◽

Viability Assay ◽

Intelligent Implant ◽

L929 Mouse

AbstractFor the development of intelligent implant systems hydrogels (HG) from crosslinked ionic liquids feature a high potential to be utilised as a drug depot. Biocompatibility of the HGs is one key prerequisite for biomedical applications. HGs were polymerised from a variety of different ionic monomers based on methacrylate, methacrylamide, styrene or vinyl imidazolium derivatives in aqueous solution. N,N'-methylenebisacrylamide was used as crosslinker. CellQuanti-Blue™ Cell Viability Assay Kit was implemented to proof viability of L929 mouse fibroblasts. The predominant part of the HG eluates generated only a marginal reduction of less than 15% cell viability at 100% eluate concentration. This underlines the excellent suitability of these HGs for biomedical applications and revealed some promising candidates for the development of drug depots for implants.

Download Full-text