Significant Decrease In Cellular Viability Research Articles

Magnetic Thermosensitive Liposomes Loaded with Doxorubicin.

Liposome-mediated anticancer drug delivery has the advantage of limiting the massive cytotoxicity of chemotherapeutic agents. Doxorubicin (DOX) PEG-liposomal does however have a slow-release rate that hinders its therapeutic efficacy. In this study, an integrated therapeutic system based on magnetic thermosensitive liposomes was designed. The chelated gadolinium acquired magnetic properties in the liposomes. The hyperthermia induced by ultra-high-field magnetic resonance imaging (UHF-MRI) enhances the chemotherapeutic effects of DOX. The DOX release from liposomes was facilitated over a narrow range of temperatures owing to the phase transition temperature of the liposomes. The magnetic properties of the liposomes were evident by the elevation of contrast after the exposure to UHF-MRI. Moreover, triple-negative breast cancer (TNBC) cells showed a significant decrease in cellular viability reaching less than 40% viability after 1h of exposure to UHF-MRI. The liposomes demonstrated a physiological coagulation time and a minimal hemolytic potential in hemocompatibility studies; therefore, they were considered safe for physiological application. As a result, magnetic-thermosensitive liposomal guidance of local delivery of DOX could increase the therapeutic index, thereby reducing the amount of the drug required for systemic administration and the chance of affecting the adjacent tissues.

Synthesis and Identification of New Heterocyclic Compounds from Isatin and Study their Inhibition Ability of Colorectal Adenocarcinoma

Heterocyclic compounds showed a wide range of applications but in medicinal chemistry are of particular interest. Heterocyclic were the active compounds in many natural drugs also some are still used today, for instance, quinine derivatives and morphine. Colorectal carcinoma is the world's third most common neoplasm and the second leading cause of cancer-related death. This research aims to identify a prospective medication with a high affinity for treating colorectal cancer. A Series of heterocyclic compounds 1, 2, 3, and 4 were synthesized and confirmed through the use of different spectroscopic techniques (IR and NMR). The in vitro their anticancer potential by means of cytotoxicity and selectivity determination of the compounds 1a, 2b, 3a, and 4a against HT-29 and MA104 were assessed. The experimental Isatin derivative 2b has a reasonable and strong antineoplastic effect on colorectal cancer HT-29 cell line, as evidenced by its significant decrease in cellular viability and increase affinity, according to our findings. In this study, 2b also showed considerable safety qualities based on the parameters tested on the Vero cell line.

Ecotoxic, genotoxic, and cytotoxic potential of leachate obtained from chromated copper arsenate-treated wood ashes.

Preservative treatments increase the durability of wood, and one of the alternative treatments involves the use of chromated copper arsenate (CCA). Due to the toxicity of CCA, the disposal of CCA-treated wood residues is problematic, and burning is considered to provide a solution. The ecotoxicological potential of ash can be high when these components are toxic and mutagenic. The aim of this study was to evaluate the toxicity and genotoxicity of bottom ash leachates originating from CCA-treated wood burning. Physical-chemical analysis of the leachates revealed that in treated wood ashes leachate (CCA-TWBAL), the contents of arsenic and chromium were 59.45 mg.L-1 and 54.28 mg.L-1, respectively. In untreated wood ashes leachate (UWBAL), these contents were 0.70 mg.L-1 and 0.30 mg.L-1, respectively. CCA-TWBAL caused significant toxicity in Lactuca sativa, Allium cepa, and microcrustacean Artemia spp. (LC50 = 12.12 mg.mL-1). Comet assay analyses using NIH3T3 cells revealed that concentrations ranging from 1.0 and 2.5 mg.mL-1 increase the damage frequency (DF) and damage index (DI). According to MTT assay results, CCA-TWBAL at concentrations as low as 1 mg.mL-1 caused a significant decrease in cellular viability. Hemolysis assay analyses suggest that the arsenic and chromium leachate contents are important for the ecotoxic, cytotoxic, and genotoxic effects of CCA-TWBAL.

Mechanical characterization of bio composite films as a novel drug carrier platform for sustained release of 5-fluorouracil for colon cancer: Methodological investigation

In this study, we employed Pectin (PC) as a matrix that is hybridized with three different nucleobase (NB) units (cytosine, thymine, uracil) to generate pectin–nucleobase(PC-NB) biocomposite films stabilized through bio-multiple hydrogen bonds (BMHBs) as drug carrier for anticancer 5-Fluorouracil (5-FU). Prepared biocomposite films were characterized by Fourier Transform Infra-red Spectroscopy (FTIR), X-ray Diffraction (XRD), Thermogravimmetry Analysis (TGA) and Scanning Electron Microscope (SEM). Mechanical and sorption properties were also evaluated. In vitro drug release performed in both acidic pH 1.2 (stomach pH) and alkaline pH 7.4 (intestinal pH) showed that incorporation of nucleobases into pectin significantly restricted release rate of 5-FU particularly under acidic condition (pH 1.2). Hemolysis assays demonstrated that PC-NB-5-FU biocomposite film drug carriers were hemocompatible. Confocal microscope analysis indicates facilitated cellular uptake of PC-NB-5-FU film in HT-29 colon cancer cell line, which in turn result in a higher potential of apoptosis. Confocal imaging of fluorescent live/dead cell indicators and MTT assay outcomes, both demonstrated significant decreases in cellular viability of PC-NB-5-FU biocomposite films. Collectively, our findings indicate that this PC-NB-5-FU biocomposite films can be conferred as a proficient formulation for targeted delivery of colon cancer drugs.

Molybdenum cluster loaded PLGA nanoparticles as efficient tools against epithelial ovarian cancer

In this study, poly (lactic-co-glycolic) acid nanoparticles loading inorganic molybdenum octahedral cluster were used for photodynamic therapy (PDT) of ovarian cancer.Three cluster compounds, ((C4H9)4N)2[{Mo6Br8}Br6], Cs2[{Mo6Br8}Br6] and Cs2[{Mo6I8}(OOC2F5)6] denoted TMB, CMB and CMIF were studied following their incorporation in nanoparticles by a nanoprecipitation method. All resulting nanoparticles exhibited physico-chemical characteristics such as size and zeta potential compatible with cellular uptake. All cluster compounds tested were shown to produce singlet oxygen in vitro once released from their nanoparticulate system. Confocal images showed an internalisation of cluster loaded nanoparticles (CNPs) in A2780 ovarian cancer cell line, more efficient with CMIF compared to CMB or TMB loaded nanoparticles. In vitro cellular viability studies conducted on A2780 cell line treated with non activated CNPs did not show any sign of toxicity for concentrations up to 15 µM. Following photo-activation, CNPs were able to generate singlet oxygen resulting in a decrease of the cellular viability, compared to non-activated conditions. Nevertheless, no significant differences between IC50 with or without photo-activation were observed with TMB and CMB CNPs while for CMIF loaded nanoparticles, the photo-activation led to a significant decrease of cellular viability compared to the non activated condition and this decrease was independant of the P/C ratio. The strong photo-toxicity obtained for CMIF loaded nanoparticles with a P/C ratio of 2.5, as shown with half maximal inhibitory concentration (IC50) value near 1.8 µM suggests that PLGA nanoparticles seem to be efficient delivery systems intended for tumor management and that CMIF can be further investigated as photosensitizer for PDT of ovarian cancer.

Effect of Combined K. pinnata and Metformin Preparation on Inflammatory Cytokines in Normal and Diabetic Skeletal Muscle Cells.

Diabetes Mellitus is associated with systemic inflammation and oxidative stress, which may play a central role in the development of diabetic complications. In this study, combined preparations of Kalanchoe pinnata and metformin were investigated to determine the effects on inflammatory activity in human skeletal muscle myoblasts (HSMMs) and human diabetic skeletal muscle myoblasts (DHSMMs). Results showed that combinatorial preparations sustained cell viability for 3 days in both HSMM and DHSMM cells. However, a significant decrease in cellular viability occurred for both cell lines on day 5. Results also indicate that combinatorial preparations of K. pinnata may modulate immune responses by significantly upregulating proinflammatory markers, interleukin (IL) 2, and tumor necrosis factor-alpha, and upregulating the anti-inflammatory marker, IL-10, in HSMM and DHSMM cells. The combined preparations significantly downregulated the anti-inflammatory glycoprotein IL-6 in both diabetic and nondiabetic human skeletal muscle cells. The findings suggest that combined preparations of K. pinnata and metformin might be a potential immune-modulating agent that may promote inflammation and adversely affect the outcome of diabetic patients.

Selenium and L-carnitine protects from valproic acid-Induced oxidative stress and mitochondrial damages in rat cortical neurons

Oxidative stress and mitochondrial dysfunction have been associated with valproic acid (VPA) induced neurotoxicity. Mitochondria are vulnerable to oxidative damage and are also a major source of superoxide free radicals. Therefore, the need for mitochondrial protective and antioxidant agents for reducing valporic acid toxicity in central nerve system (CNS) is essential. In the present study, we investigated the potential beneficial effects of sodium selenite (SS) and L-carnitine (LC) against valproic acid -induced oxidative stress and mitochondrial dysfunction in isolated rat cortical neurons. Valproic acid (50, 100 and 200 µM) treatment caused a significant decrease in cellular viability, which was accompanied by increases in reactive oxygen species (ROS) generation, GSSG and GSH content, lipid peroxidation and lysosomal and mitochondrial damages. Sodium selenite (1 µM) and L-carnitine (1 mM) pretreatment attenuated valproic acid-induced decrease in cell viability. In addition, sodium selenite (1 µM) and L-carnitine (1 mM) pretreatment significantly protected against valproic acid-induced raise in oxidative stress, mitochondrial and lysosomal dysfunction, lipid peroxidation levels and depletion of GSH content. Our results in the current study provided insights into the protective mechanism by L-carnitine and sodium selenite, which is liked, to neuronal ROS generation and mitochondrial and lysosomal damages.

Effects of photobiomodulation on cellular viability and cancer stem cell phenotype in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is the most common head and neck malignancy; it has been shown that cancer stem cells (CSC) are present in OSCC and associated with tumor growth, invasion, metastasis, and therapeutic resistance. Photobiomodulation (PBM) is an alternative tool for oncologic treatment adverse effects such as oral mucositis (OM); however, controversy exists regarding the undesirable effects of PBM on tumor or CSC. This study aimed to evaluate in vitro, the effects of PBM, with the same dosimetric parameters as those used in the clinic for OM prevention and treatment, on OSCC cellular viability, as well as PBM's effect on CSC properties and its phenotype. OSCC cell lines were submitted to single or daily PBM with 3 J/cm2 and 6 J/cm2 and then the cellular viability was evaluated by MTT, NRU (neutral red uptake), and CVS (crystal violet staining). The CSC populations were evaluated by clonogenic formation assay, flow cytometry, and RT-qPCR. The single PBM with the 3 J/cm2 group was associated with increased cellular viability. Daily PBM with 3 J/cm2 and 6 J/cm2 was associated with a significant decrease in cellular viability. Additionally, daily PBM was not able to promote CSC self-renewal or the CD44high/ESAlow and CD44high/ESAhigh cellular phenotypes. Moreover, a decrease in the number of spheres and in the expression of the CSC related gene BMI1 was observed after daily PBM with 6 J/cm2. Daily PBM with 3 J/cm2 and 6 J/cm2 showed an inhibitory effect on cellular viability and was not able to promote the CSC self-renewal or phenotype.

Trifluoperazine an Antipsychotic Drug and Inhibitor of Mitochondrial Permeability Transition Protects Cytarabine and Ifosfamide-Induced Neurotoxicity.

The link between Ca2+ dysregulation, mitochondria damages, oxidative stress and cellular derangement is particularly evident in neurotoxicity induced by chemotherapeutic agents. In the current study, we investigated effects of trifluoperazine (TFP) as an inhibitor of calmodulin against the cytotoxicity induced by cytarabine (Ara-C) and Ifosfamide (IFOS) on isolated rat neurons and also the mechanisms involved in this toxicity. Isolated rat neurons were pretreated with TFP (100 µM) for 5 min at 37°C, then Ara-C (226 µM) and IFOS (290 µM) were added in separate experiments. After 3 h, the cytotoxicity, reactive oxygen species (ROS), lysosomal membrane destabilization, mitochondrial membrane potential (MMP), lipid peroxidation (LP), glutathione (GSH) and glutathione disulfide (GSSG) levels were measured. Ara-C and IFOS treatments caused a significant decrease in cellular viability, which was accompanied by ROS generation, GSSG/GSH ratio, lipid peroxidation and lysosomal and mitochondrial damages. On the other hand, TFP (100 µM) pre-treatment attenuated Ara-C and IFOS -induced decrease in cell viability. In addition, TFP (100 µM) pre-treatment significantly protected against Ara-C and IFOS -induced increase in ROS generation, lysosomal and mitochondrial damages, lipid peroxidation levels and decrease in GSH/GSSG ratio. Our data provided insights into the mechanism of protection by TFP against Ara-C and IFOS neurotoxicity, which is related, to neuronal ROS formation and mitochondrial damages.

Cytotoxic Activity of Epigallocatechin and Trans-Cinnamaldehyde in Gastric Cancer Cell Line

Introduction: Herbal medication is currently being utilized for treatment of numerous diseases, such as cancer which showed successful therapeutic efficacy in numerous studies. Epigallocatechin gallate (EGCG) is a compound of green tea that its role in tumor cell death has been reported. Likewise, trans-cinnamaldehyde (TC), the active ingredient in cinnamon oil showed anti-cancer properties in some previous studies. The aim of this study was to evaluate the cytotoxic effects of EGCC and TC on proliferation of gastric cancer cell line (AGS).Methods: AGS Cells were seeded and treated with various concentrations of EGCG and TC for 72 h and assessed for cell viability. To study the cytotoxic effect of drug in combination cases the lower doses than IC50 of EGCG and TC was utilized. The one-way analysis of variance (ANOVA) were used for statistical analysis.Results: various concentrations of EGCG and TC significantly inhibited the proliferation of AGS cells in dose dependent pattern. We found that in double combined cases cellular viability decreased in compared to IC50 of each single agents. Also, there were significant decrease in cellular viability in all single and double treated cases toward untreated cells (p<0.05).Conclusion: The results of this study indicated that EGCG and TC effects on AGS cell line were significantly high and dose-dependent and might be cooperative. Double combinations of these two agents may be considered as a potential therapeutic option for gastriccancer. Further investigation should be conducted to validate these combination in gastric cancer therapy.

Sodium selenite protects from 3-nitropropionic acid-induced oxidative stress in cultured primary cortical neurons.

Selenium (Se) is an essential trace element for humans; its intake is needed to allow the proper synthesis of 25 different selenoproteins that are necessary to the normal functioning of several organs, including the brain. Accordingly, decreased Se levels have been associated with neurological disorders. In the present study, we investigated the potential beneficial effects of Se, as sodium selenite, against 3-nitropropionic acid (3-NP)-induced oxidative stress in primary cultures of mouse cortical neurons. 3-NP treatment caused a significant decrease in cellular viability, which was accompanied by decreases in mitochondrial complex II activity and reduced glutathione (GSH) content, as well as increases in reactive oxygen species (ROS) generation and oxidized glutathione (GSSG) levels. Sodium selenite pretreatment (6days) attenuated 3-NP-induced decrease in cell viability. In addition, sodium selenite pretreatment significantly protected against 3-NP-induced increase in ROS generation and decrease in GSH/GSSG ratio. Of note, sodium selenite pretreatment did not change 3-NP-induced decrease of mitochondrial complex II activity, suggesting that Se modulates secondary events resultant from 3-NP-induced mitochondrial dyshomeostasis. In addition, sodium selenite pretreatment significantly increased glutathione peroxidase (GPx) activity. Our data provide insights into the mechanism of protection by sodium selenite, which is related, at least in part, to GPx induction.

The effects of doxorubicin-loaded liposomes on viability, stem cell surface marker expression and secretion of vascular endothelial growth factor of three-dimensional stem cell spheroids.

The aim of the present study was to evaluate the effects of anionic, cationic and neutral liposomes containing doxorubicin on the cellular viability and osteogenic differentiation of three-dimensional stem cell spheroids. Doxorubicin-loaded liposomes were prepared using the traditional thin-lipid-film-hydration method and were characterized using transmission electron microscopy and a zeta potential analyzer. The doxorubicin release profile from these liposomes was also analyzed in vitro. Three-dimensional cell spheroids were fabricated using silicon elastomer-based concave microwells. Qualitative results of cellular viability were observed using a confocal microscope and quantitative cellular viability was evaluated using a Cell-Counting Kit-8 (CCK-8) assay. Furthermore, the secretion of vascular endothelial growth factor was evaluated. Western blot analysis was performed to assess the expression of collagen I and glyceraldehyde 3-phosphate. Results indicated that the spheroids were well formed in silicon elastomer-based concave microwells on day 1. In general, the shapes of the cells in the in the doxorubicin-loaded anionic, cationic and neutral liposome groups were similar to the control group except for the 10 µg/ml groups on days 3, 5, and 7. No significant changes in cellular viability were noted with the addition of doxorubicin at day 1 but significant decreases in cellular viability were noted with application of doxorubicin at day 5. Notably, higher concentrations of doxorubicin reduced the secretion of vascular endothelial growth factor and stem cell marker expression. To conclude, the present study indicated that doxorubicin-loaded anionic liposomes produced the most sustained release profile and cationic liposomes produced the highest uptake of the stem cell spheroids. These findings suggested that higher concentrations of doxorubicin-loaded liposomes affected cellular viability, the secretion of vascular endothelial growth factor and stem cell marker expression.

Abstract 512: Knockdown of the spindle assembly checkpoint gene Bub1b results in increased cell death and cell cycle impairment in breast cancer

Abstract Genomic instability is an enabling hallmark of cancer that provides cancer cells a replicative advantage. Accumulation of genomic aberrations can also compromise the genomic integrity and put cells under mitotic stress. Maintaining balance between the instability that gives cancer cells a replicative advantage and the instability that could lead them to mitotic catastrophe is crucial for survival of cancer cells. Mitotic catastrophe can be caused by rapid progression through mitosis before crucial checkpoints are met, resulting in cell death. To overcome this challenge, cancer cells may acquire overexpression of spindle assembly checkpoint (SAC) genes, which can prevent mitotic catastrophe that would occur if they undergo mitosis prematurely. Bub1b is an important part of SAC and inhibits the onset of anaphase until all chromosomes are aligned at the metaphase plate. Our analysis of clinical datasets show Bub1b expression is elevated in breast cancer, compared to normal breast, and is exemplified by a pattern of increasing overexpression in more aggressive variants, such as triple negative breast cancer (TNBC). Bub1b overexpression also correlates with decreased overall survival in patients. Expression analysis of breast cancer cell lines corroborates this clinical data. We hypothesize that the requirement for Bub1b expression indicates a vulnerability of rapidly proliferating breast cancers cells, and the inhibition of Bub1b will result in cell death through mitotic catastrophe. Using RNA interference with siRNAs we knocked down Bub1b expression in a variety of breast cancer cells. This resulted in significant decrease in cellular viability and clonogenicity in soft agar. Furthermore, analyses of apoptosis using Annexin V and PI costaining showed that Bub1b knockdown results in significantly increased apoptosis and cell death, especially in the TNBC cell line MDA-MB-468. However, the viability and apoptosis levels of the normal breast epithelial cell line, MCF12A, were not affected. Bub1b knockdown resulted in silencing of SAC activity, as measured by Cyclin B levels, and also significantly impaired cell cycle progression in breast cancer cell lines. Additionally, knockdown of Bub1b in breast cancer cell lines demonstrated an additive effect when combined with DNA-damaging agents. Our data support that Bub1b is a critical player in breast cancer cell survival, by enabling them to overcome mitotic stress. We demonstrate that impairment of Bub1b has detrimental effects on cell cycle and progression of mitosis. Further investigation of the role of Bub1b in promoting successful chromosome alignment and proliferation of breast cancer cells with genomic instability could provide a novel pathway on how cancer overcomes the mitotic stress caused by genomic instability. These studies may also lead to novel therapeutic strategies that combine SAC inhibitors with standard genotoxic treatments. Citation Format: Dilara Koyuncu, Erik T. Goka, Philip C. Miller, Marc E. Lippman. Knockdown of the spindle assembly checkpoint gene Bub1b results in increased cell death and cell cycle impairment in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 512. doi:10.1158/1538-7445.AM2017-512

The Effect of Silica Nanoparticles on Human Corneal Epithelial Cells.

Ocular drug delivery is an interesting field in current research. Silica nanoparticles (SiNPs) are promising drug carriers for ophthalmic drug delivery. However, little is known about the toxicity of SiNPs on ocular surface cells such as human corneal epithelial cells (HCECs). In this study, we evaluated the cytotoxicity induced by 50, 100 and 150 nm sizes of SiNPs on cultured HCECs for up to 48 hours. SiNPs were up-taken by HCECs inside cytoplasmic vacuoles. Cellular reactive oxygen species generation was mildly elevated, dose dependently, with SiNPs, but no significant decrease of cellular viability was observed up to concentrations of 100 μg/ml for three different sized SiNPs. Western blot assays revealed that both cellular autophagy and mammalian target of rapamycin (mTOR) pathways were activated with the addition of SiNPs. Our findings suggested that 50, 100 and 150 nm sized SiNPs did not induce significant cytotoxicity in cultured HCECs.

Graphene oxide induces apoptotic cell death in endothelial cells by activating autophagy via calcium-dependent phosphorylation of c-Jun N-terminal kinases.

Graphene oxide (GO) have received considerable attention with respect to their utilization in biomedical applications. However, GO-related safety issues concerning human vasculature are very limited. In this manuscript, we report for the first time the differential size-related biological effects of GOs on endothelial cells (ECs). Notably, Subnanometer- and nanometersized GOs induce apoptotic death in ECs via autophagy activation. We propose a molecular mechanism for the GO-induced autophagic cell death through the PLCβ3/IP3/Ca2+/JNK signaling axis. Our findings could be provide a better understanding of the GO sizedependent cytotoxicity in vasculature and facilitate the future development of safer biomedical applications of GOs.

Gold Nanoantenna-Mediated Photothermal Drug Delivery from Thermosensitive Liposomes in Breast Cancer.

In this work, we demonstrate controlled drug delivery from low-temperature-sensitive liposomes (LTSLs) mediated by photothermal heating from multibranched gold nanoantennas (MGNs) in triple-negative breast cancer (TNBC) cells in vitro. The unique geometry of MGNs enables the generation of mild hyperthermia (∼42 °C) by converting near-infrared light to heat and effectively delivering doxorubicin (DOX) from the LTSLs in breast cancer cells. We confirmed the cellular uptake of MGNs by using both fluorescence confocal Z-stack imaging and transmission electron microscopy (TEM) imaging. We performed a cellular viability assay and live/dead cell fluorescence imaging of the combined therapeutic effects of MGNs with DOX-loaded LTSLs (DOX-LTSLs) and compared them with free DOX and DOX-loaded non-temperature-sensitive liposomes (DOX-NTSLs). Imaging of fluorescent live/dead cell indicators and MTT assay outcomes both demonstrated significant decreases in cellular viability when cells were treated with the combination therapy. Because of the high phase-transition temperature of NTSLs, no drug delivery was observed from the DOX-NTSLs. Notably, even at a low DOX concentration of 0.5 μg/mL, the combination treatment resulted in a higher (33%) cell death relative to free DOX (17% cell death). The results of our work demonstrate that the synergistic therapeutic effect of photothermal hyperthermia of MGNs with drug delivery from the LTSLs can successfully eradicate aggressive breast cancer cells with higher efficacy than free DOX by providing a controlled light-activated approach and minimizing off-target toxicity.

Effect of topical neuromodulatory medications on oral and skin keratinocytes.

Neuromodulatory medications (NMs), such as amitriptyline, carbamazepine and gabapentin, are used as topical preparations for the management of neuropathic orofacial pain (NOP) and have produced promising preliminary results. The aim of this study was to investigate the effects of three aforementioned NMs on cell lines relevant to the orofacial tissues in vitro as no published studies have examined the effect of these topical NMs. Cellular viability was measured using alamarBlue® , testing cumulative and specific time point effects of NMs on human skin keratinocytes and oral keratinocytes. Effects of the NMs on cell counts were investigated by CCK-8 assay. Drug concentrations released from NM orabase pastes after 30-min incubation were measured by high-performance liquid chromatography. Using these clinical concentrations, morphological changes and cytokine expression were investigated using scanning electron microscopy (SEM) and human inflammatory antibody array (AAH), respectively. Cumulative and specific time point viability and cell count methods revealed that amitriptyline caused a significant decrease in cellular viability and counts in both cell lines. Carbamazepine also had significant effects after long-term exposure and at higher concentrations, whilst gabapentin had little demonstrable effect. SEM confirmed the cytotoxicity of amitriptyline, whilst AAH revealed no significant changes in cytokine expression following amitriptyline, carbamazepine or gabapentin exposure compared with control. The results raise concerns about the safety of topical amitriptyline as it was cytotoxic to skin and oral keratinocytes in both exposure times and concentrations, whilst carbamazepine was cytotoxic only at high concentrations and after longer exposure times and gabapentin had no demonstrable effects.

Evaluation of cytotoxic and apoptotic effects of several homeopathic dilutions of Echinacea angustifolia on human breast, cervical and prostate cancer cells and genotoxic study of E. angustifolia MT

Introduction : Cancer research needs a multi-disciplinary approach from several areas of medicine. The use of homeopathic drugs in cancer treatment has generated wide interest and controversy. Objective : The aim of this research was to study the efficacy of homeopathic dilutions of Echinacea angustifolia on different cancer cell lines. Methods : Cytotoxic activity of E. angustifolia mother tincture (MT) and homeopathic dilutions (6C, 30C, 200C, and 1M) was assessed on human cell lines: MDA-MB-231 (Breast cancer), HeLa (Cervical cancer) and PC-3 (Prostate cancer). Cytotoxicity was measured using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) method. Apoptosis was determined by Annexin-V with flow cytometry. To demonstrate the safety of homeopathic dilutions, cytotoxicity was measured on peripheral blood mononucleated cells (PBMC). Genotoxicity was evaluated by Ames test (in vitro) and Micronucleus assay (in vivo). Results : E. angustifolia homeopathic dilutions had a statistically significant decrease of cellular viability (p Conclusion : This study provides scientific evidence of the ability of E. angustifolia homeopathic dilutions to induce apoptosis in the breast cancer cell line MDA-MB-231, which encourages a possible use as supportive medicines in cancer therapy. Further in vivo studies of these homeopathic remedies must be performed.

Nanosecond pulsed electric fields (nsPEFs) impact and enhanced Photofrin II® delivery in photodynamic reaction in cancer and normal cells

BackgroundNanosecond pulsed electric field (nsPEF) treatment is a new anti-cancer strategy with ultrashort pulse duration and high intensity of electric fields. The application of nsPEFs affects all intra- and extracellular membranes and independently initiates the process of apoptosis within cancer cells, leading the tumor to slowly auto-destruct without the use of toxic drugs. MethodsThis study involves cells of gastric adenocarcinoma (EPG85-257P and EPG85-257RDB), metastatic melanoma (Me45), epidermal cancer (A431), normal keratinocytes (HaCaT), and macrophages (P388/D1). The influence of nanosecond pulses on the cellular structure and cellular proliferation was evaluated. The effect of nsPEF was determined by MTT and clonogenic assays and the efficiency was monitored by following the propidium iodide and Photofrin II® uptake using FACS analysis. The cell membranes state was visualized with DHCC marker. ResultsnsPEFs (up to 60kV/cm) induced significant decrease of cellular viability in all cancer cells except the A431 cell line. Photodynamic reactions combined with nsPEFs induced the highest decrease of cellular viability in both gastric cell lines and skin derived cancer cells. Normal (HaCaT and P388/D1) cells were in contrary not significantly affected. Propidium iodide and Photofrin II® uptake, used as markers of membrane permeabilization, were the most efficient in gastric cells. Finally, the most disturbed morphology was observed in the latter. ConclusionsThis is the first attempt of combining nsPEF with photodynamic reaction using Ph II® for selective destruction of cancer cells. The results indicate the potential of nsPEF for inducing cytotoxicity mainly in adenocarcinoma cells, while combined with Photofrin II® and irradiation.

4-hydroxynonenal regulates mitochondrial function in human small airway epithelial cells.

Prolonged exposure to oxidative stress causes Acute Lung Injury (ALI) and significantly impairs pulmonary function. Previously we have demonstrated that mitochondrial dysfunction is a key pathological factor in hyperoxic ALI. While it is known that hyperoxia induces the production of stable, but toxic 4-hydroxynonenal (4-HNE) molecule, it is unknown how the reactive aldehyde disrupts mitochondrial function. Our previous in vivo study indicated that exposure to hyperoxia significantly increases 4-HNE-Protein adducts, as well as levels of MDA in total lung homogenates. Based on the in vivo studies, we explored the effects of 4-HNE in human small airway epithelial cells (SAECs). Human SAECs treated with 25 μM of 4-HNE showed a significant decrease in cellular viability and increased caspase-3 activity. Moreover, 4-HNE treated SAECs showed impaired mitochondrial function and energy production indicated by reduced ATP levels, mitochondrial membrane potential, and aconitase activity. This was followed by a significant decrease in mitochondrial oxygen consumption and depletion of the reserve capacity. The direct effect of 4-HNE on the mitochondrial respiratory chain was confirmed using Rotenone. Furthermore, SAECs treated with 25 μM 4-HNE showed a time-dependent depletion of total Thioredoxin (Trx) proteins and Trx activity. Taken together, our results indicate that 4-HNE induces cellular and mitochondrial dysfunction in human SAECs, leading to an impaired endogenous antioxidant response.