Normal Mouse Fibroblasts Research Articles

Tin (IV) oxide (SnO2) nanoparticles inhibit the viability of cervical cancer HeLa cells through induction of apoptosis

Introduction: Resistance to chemotherapy and severe side effects have been reported as the main reasons for treatment failure in patients with cervical cancer. Therefore, it is necessary to find new treatment strategies with fewer side effects and more efficacy. This study aimed to investigate the cytotoxic property of tin (IV) oxide (SnO2) nanoparticles (NPs) against human cervical cancer cells (HeLa cells). In addition, the molecular mechanism of anticancer activity of SnO2 NPs was evaluated. Materials and Methods: The cytotoxicity of SnO2 NPs against HeLa cells and normal mouse fibroblast cells (L929) was studied using an MTT assay. To determine the mechanism of action of SnO2 NPs, the cells were treated with the half maximal inhibitory concentration values of SnO2 NPs for 24 h and apoptotic cell percentage was assessed by Annexin-PI and flow cytometry. In addition, real-time quantitative polymerase chain reaction (PCR) was used to evaluate the mRNA expression levels of apoptotic genes (Bax and Bcl-2). Results: SnO2 NPs suppress the viability of HeLa cells in a dose-dependent manner. This compound was more cytotoxic against HeLa cells than L929 cells. Flow-cytometry analysis revealed that SnO2 NPs significantly caused cell growth arrest. Moreover, real-time PCR results showed that SnO2 NPs treatment decreased Bcl-2 and increased Bax expression level. Conclusion: SnO2 NPs treatment significantly inhibit HeLa cells viability through the induction of apoptosis. Interestingly SnO2 NPs were more cytotoxic against HeLa cells than normal fibroblast cells, which may provide promising evidence for their applications as an anticancer drug.

Anticancer Activity of Ethnopharmacological Plants of Golestan Province/Iran against AGS, HT-29 and KYSE-30 Cell Lines through Promoting the Apoptosis and Immunomodulatory Effects.

The anticancer and immunomodulatory effects of medicinal plants from Golestan province, as a promising source of cancer therapy against gastrointestinal cancer cell lines, were investigated in this study. The ethanolic root/aerial part extracts of 9 medicinal plants were screened for their cytotoxicity against normal mouse fibroblast cells (L-929) and three human cancer cell lines including gastric adenocarcinoma (AGS), colorectal adenocarcinoma (HT-29), and esophagus adenocarcinoma (KYSE-30) by performing MTT assay to determine the IC50 of the extracts. The in-vitro antioxidant activity, total phenolic (TPC), and total flavonoid content (TFC) of extracts was evaluated. Flow cytometry and Real-Time PCR were used for apoptosis assay and evaluation of expression of some genes involved in cell signaling; TLR-4, AKT, ERK1/2, and NFκB. Out of the 9 plant extracts screened, Arctiumlappa root (ALR), showed the most potent cytotoxicity against AGS, KYSE-30, and HT-29 cells with IC50 values of 10, 200, and 2030 µg/mL, respectively. In addition, ALR exerts high TPC (215.8 ± 0.3 mg GAE/g), TFC (69.03 ± 0.7 mg QUE/g) and high radical scavenging activity with IC50 (1250 ‎±‎ 0.1 µg/mL) in DPPH method. Also, ALR stimulates TLR-4 signaling, increased apoptosis, and decreased cancer cell attachment to the surface compared to the untreated cells. This plant, with a strong cytotoxic effect on cancer cells as well as increased apoptosis and its effect on molecules involved in TLR4 signaling as the immunomodulatory effect can be a suitable candidate for in-vivo studies in the future for cancer therapy.

Synthesis, crystal structure and Hirshfeld Surface analysis of benzamide derivatives of thiourea as potent inhibitors of α-glucosidase in-vitro

Benzamide based structural analogues 1–15 were synthesized, and evaluated for α-glucosidase inhibition activity in vitro for the first time. Compounds 1–9 were found to be known, while compounds 10–15 were found to be new. However, to the best of our knowledge we are reporting α-glucosidase inhibitory activity of these bezamide derivatives of thiourea for the first time. Compounds 1, 3, 6–8, 10–14 were found to be potent inhibitors of α-glucosidase within IC50 range of 20.44–333.41 µM, in comparison to the standard inhibitor, acarbose (IC50 = 875.75 ± 2.08 µM). Mode of the enzyme inhibition was determined on the basis of kinetic studies which demonstrated that compounds 8, and 10 were non-competitive and competitive inhibitors of α-glucosidase enzyme, respectively. These compounds were also evaluated for their DPPH radical scavenging activity, and cytotoxicity against 3T3 mouse fibroblast cell lines. All synthesized compounds showed a significant to moderate DPPH radical scavenging activity and appeared to be non-cytotoxic except compound 9 which showed cytotoxicity against 3T3 normal mouse fibroblast cell lines. A single crystal X-ray and Hirshfeld Surface analysis of a representative compound is also presented.

Cytotoxic Activity and Initiation of Apoptosis via Intrinsic Pathway in Jurkat Cells by Leaf Extract of Zanthoxylum rhetsa DC

Background Newer drugs are in demand for leukemia treatment that specifically targets tumor cells without affecting normal cells. Potent cytotoxic activities have been reported from various parts of Zanthoxylum rhetsa. Thus, the present study was conducted to evaluate antileukemic potential of leaf extract of Z. rhetsa along with probable mechanism of cytotoxicity. Materials and Methods: The antiproliferative activity of the extract on leukemic cell lines was evaluated using sulforhodamine B assay. The changes in cell death profile, cell cycle, and expression levels of pro-apoptotic markers (p53, Bax, cytochrome C, caspase 3, and MMP) and antiapoptic marker (Bcl2) on Jurkat cell lines were studied using flow cytometer. Comparison of oxidative stress induced by extract on Jurkat cells and normal mouse fibroblast cells was done. DNA fragmentation was studied using gel electrophoresis. Results: The leaf extract showed concentration-dependent cytotoxicity against Jurkat cell lines majorly via apoptotic mechanism. It arrested cells at G0/G1 and S phase of cell cycle. Apoptosis was associated with increase in the expression of pro-apoptotic markers and decrease of anti-apoptotic markers. The treatment with extract selectively increased the oxidative stress in Jurkat cells and showed DNA fragmentation. Conclusion: The methanol extract of leaves of Z. rhetsa show selective cytotoxic activity on Jurkat cell lines and induced apoptosis via intrinsic pathway.

Induction of mitochondria-mediated apoptosis and suppression of tumor growth in zebrafish xenograft model by cyclic dipeptides identified from Exiguobacterium acetylicum

Colorectal cancer is the most common type of gastrointestinal cancers with poor survival and limited therapeutic options. In this study, four structurally different cyclic dipeptides (or diketopiperazine) were isolated and identified as cyclo (l-Pro-l-Leu), cyclo (l-Pro-l-Val), cyclo (l-Pro-l-Phe) and cyclo (l-Pro-l-Tyr) from the ethyl acetate extract in the cell-free filtrate of Exiguobacterium acetylicum S01. The anticancer potential of identified DKPs on colorectal cancer HT-29 cells in vitro and in vivo zebrafish xenograft model was evaluated. The MTT (3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide)) assay showed that four DKPs exhibited significant inhibition of HT-29 cells viability in a dose-dependent manner whereas there were no cytotoxic effects on normal mouse fibroblast 3T3 cells. Also, we observed that all DKPs induce early and late apoptotic cell death in HT-29 cells. Moreover, the expression levels of apoptotic (cytochrome-c, caspase-3 and Bid) and anti-apoptotic (Bcl-2) markers were up- and down-regulated in HT-29 cells in response to DKPs treatments. Furthermore, these four DKPs remarkably inhibited the tumor progression in a zebrafish xenograft model within a nonlethal dose range. Overall, our findings suggest that cyclic dipeptides derived from E. acetylicum S01 could be promising chemopreventive/ therapeutic candidates against cancer.

Preparation, characterization and anti-cancer activity of graphene oxide‐‑silver nanocomposite

This work reported the preparation, characterization, cytotoxicity of green synthesized Lespedeza cuneate mediated silver nanoparticles (Lc-AgNPs) and graphene oxide‑silver nanocomposite (GO-AgNComp) using Lc-AgNPs. The UV absorption spectrum at 419 nm indicated the successful formation of GO-AgNComp. The TEM analysis displayed the thin sheet of graphene decorated Lc-AgNPs in GO-AgNComp. Zeta potential was −13.2 mV for Lc-AgNPs and −30.5 mV for GO-AgNComp. The photothermal conversion efficiency was calculated as 31.09% for GO-AgNComp. The negatively charged zeta potential of GO-AgNComp enhanced its cellular penetration through enhanced permeability and retention (EPR) effect. The near-infrared laser (NIR) induced the anticancer activity of Lc-AgNPs and GO-AgNComp in human lung cancer cells (A549) and brain tumour (LN229). The results indicated that about 50% of A549 cells and LN229 cells were ablated by treatment of 24.73 ± 2.98 μg/mL and 27.34 ± 1.62 μg/mL of Lc-AgNPs, as well by 15.46 ± 2.31 μg/mL and 20.95 ± 1.35 μg/mL of GO-AgNComp respectively. Moreover, GO-AgNComp was not cytotoxic to normal mouse fibroblast cells (NIH3T3), but it caused the cancer cell death in A549 and LN229 through ROS generation, nuclear damage, and mitochondrial membrane potential (∆ψm) loss. This work reported the anticancer potential of GO-AgNComp, which deserves further study on the molecular elucidation of GO-AgNComp mediated human lung and tumour therapy.

Type I Collagen Suspension Induces Neocollagenesis and Myodifferentiation in Fibroblasts In Vitro.

The ability of a collagen-based matrix to support cell proliferation, migration, and infiltration has been reported; however, the direct effect of an aqueous collagen suspension on cell cultures has not been studied yet. In this work, the effects of a high-concentration aqueous suspension of a micronized type I equine collagen (EC-I) have been evaluated on a normal mouse fibroblast cell line. Immunofluorescence analysis showed the ability of EC-I to induce a significant increase of type I and III collagen levels, parallel with overexpression of crucial proteins in collagen biosynthesis, maturation, and secretion, prolyl 4-hydroxylase (P4H) and heat shock protein 47 (HSP47), as demonstrated by western blot experiments. The treatment led, also, to an increase of α-smooth muscle actin (α-SMA) expression, evaluated through western blot analysis, and cytoskeletal reorganization, as assessed by phalloidin staining. Moreover, scanning electron microscopy analysis highlighted the appearance of plasma membrane extensions and blebbing of extracellular vesicles. Altogether, these results strongly suggest that an aqueous collagen type I suspension is able to induce fibroblast myodifferentiation. Moreover, our findings also support in vitro models as a useful tool to evaluate the effects of a collagen suspension and understand the molecular signaling pathways possibly involved in the effects observed following collagen treatment in vivo.

AB0175 INNOVATIVE PREPARATION OF CURCUMIN NANOPARTICLES TO IMPROVE ANTI-INFLAMMATORY EFFECT IN RHEUMATIC DISEASE

Background:Curcumin (Cur) as a natural compound can be used in the wide spectrum of healthy functions and pharmacological activities [1-4]. It shows great promise for medication of various pro-inflammatory chronic illnesses [5]. In this study, we evaluate the ability of poly(lactide-co-glycolide)(PLGA) and different grads of PVA (polyvinyl alcohol) and lecithin as a drug delivery system for poorly soluble CurObjectives:The goal of this study was to prepare and characterize Cur encapsulated PLGA and different grads of PVA and lecithin as an efficient nanocarrier for improve anti-inflammatory effect in rheumatic diseaseMethods:The PLGA nanospheres were formulated and then characterized for percent yield, encapsulation efficiency, surface morphology, and in vitro drug release profiles. At first, 6 mg of Cur was added to the organic phase including 24 mg of polymer dissolved in 5 mL of dichloromethane to constitute 1:4 (drug-to-polymer) ratios. Then, a mixture of PVA-lecithin (at about 5 cc) was added to maintain the stability of double emulsion droplets. The emulsion was continuously stirred at 300 rpm for 24 hours (at temperature of 37.5 ˚C) to evaporate the solvent, leaving behind the colloidal suspension of the drug-encapsulated nanoparticle in aqueous phase. The encapsulation of Cur into PLGA was characterized by Fourier transform infrared spectroscopy (FT-IR) and Transmission electron microscopy (TEM).Results:Our studies achieved the successful formation of smooth surface and spherical shape Cur encapsulated into PLGA nanoparticles by the TEM image confirmed. The particle size distribution demonstrated a range of 30 nm to 100 nm, with the mean particle size being 45 nm. FTIR study implies successful loading of Cur into the nanoparticles. We show high drug-loading efficiency about 98 ± 0.5% for 6% of Cur weight in total ingredients weight of PLGA (w/w). It was also seen that a slower sustained release of 10% CUR in 48 hours is observed with biocompatible PLGA in phosphate buffered saline (pH = 7.4). The MTT assay of the Cur-PLGA exhibited no cytotoxic effect on Normal mouse fibroblast cells (L-929) cell line. IC50 of Cur -PLGA increased 99.5% against Cur nanoparticles (33.57 ±0.62 µM) (P < 0.05).Conclusion:In this study, we constructed a novel preparation of curcumin nanoparticles with PLGA and different grads of PVA (polyvinyl alcohol) and lecithin to improve the bioavailability of CUR and PLGA exhibited no cytotoxic effect on L-929 cell line

Assessment of in vitro biological activities of Terminalia arjuna Roxb. bark extract and Arjunarishta in inflammatory bowel disease and colorectal cancer

Alternative or complementary therapies for several inflammatory disorders have gained considerable acceptability and popularity in recent years. The Arjuna tree, Terminalia arjuna Roxb. (Combretaceae) holds antidiarrheal and antioxidant potential useful in management of inflammatory gastro intestinal ailments. Here, we evaluated the possible effect of T . arjuna hydroalcoholic extract (TAHA) and traditional Ayurvedic formulation Arjunarishta (AA) for the treatment of inflammatory bowel disease (IBD) and colorectal cancer. The phytochemical profile of test materials was confirmed via investigation of total phenolic and flavanoid content and standardized by HPLC-PDA method. In vitro antioxidant activity was carried out using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing ability of plasma (FRAP) assay. Antimicrobial potential was tested against clinical isolates of IBD patients (HM95, HM233, HM251, HM615). Cytotoxicity was determined against human colorectal adenocarcinoma cells (Caco2, COLO.205), whereas, cytocompatibility against normal rat intestinal epithelial (IEC-6) and mouse fibroblast cells (L929). Additionally, in vitro oxidative cell damage stress was estimated by lipid peroxidation biomarker. TAHA displayed higher antioxidant capacity as compared to AA formulation. Different sensitivities were observed against different study cell lines in dose dependant manner. Similarly, significant ( p <0.05) enhanced malondialdehyde (MDA) concentrations in test materials and 5-FU treated colorectal adenocarcinoma cells was detected as compared to control cells. TAHA and AA exhibited antimicrobial activity against IBD associated clinical isolates. These findings provide biological evidence for therapeutic application of TAHA and AA in IBD and colorectal cancer treatment

Chrysin-Anchored Silver and Gold Nanoparticle-Reduced Graphene Oxide Composites for Breast Cancer Therapy

Natural polyphenol assisted fabrication of nanohybrids with improved stability and bioactivity have gained considerable interest in material science. In this study, the anticancer flavone chrysin (5,7-dihydroxyflavone ChR) was employed to fabricate silver (AgNPs) and gold nanoparticles (AuNPs) hybridized reduced graphene oxide (rGO) nanocomposites (ChR@Ag-rGONCs and ChR@Au-rGONCs) with long-term stabilities. Prepared nanocomposites (NCs) were characterized using high-throughput analyses. The hydroxyl (O–H) and carbonyl (C═O) functional groups of ChR actively contribute to the reduction of GO/metal ions (Ag+ and Au3+) and uniform decoration of Ag and AuNPs on the rGO active sites. The presence of noble metal NPs on the rGO surface improved its thermal stability and performances in diverse physiological conditions. Moreover, the surface passivation of the fabricated NCs with natural anticancer flavone ChR improved its biocompatibility. Due to the synergistic impact of the plasmonic metal–rGO hybrids, the fabricated NCs exhibited an increased cytotoxic effect over that of free-ChR against two different breast carcinoma (MDA-MD-468 and MDA-MD-231) cell lines. The prepared NCs showed comparatively less toxicity against the normal mouse fibroblast (L929) cells, demonstrating the high safety of biofunctionalized NCs in anticancer therapy. We demonstrated that the generation of reactive oxygen species released by the metal ions coexisting on the rGO induce apoptosis. Hence, this work offers a natural and viable scaling-up procedure to synthesize noble metal NPs decorated hybrid rGO composites for useful antibreast cancer applications.

Organotin (IV) derivative of Piperic acid and Phenylthioacetic acid: Synthesis, Crystal structure, Spectroscopic characterizations and Biological activities

Three new organotin (IV) derivatives have been prepared from piperic and phenylthio acetic acids, the former is obtained by hydrolysis of piperine, which is extracted from black pepper. The three complexes {[n-Bu 2 SnO 2 C-(CH=CH) 2 -C 7 H 5 O 2 ] 2 O} 2 1 , {[n-Bu 2 SnO 2 C-CH 2 -S-C 6 H 4 ] 2 O} 2 2 , and [Ph 3 SnO 2 C-(CH=CH) 2 -C 7 H 5 O 2 ] n 3 , have been characterized by IR, 1 H and 13 C NMR spectroscopic techniques. Single crystal diffraction studies were made to determine the structures of the three compounds 1 , 2 and 3 . Compounds 1 and 2 crystallize in the triclinic system ( P\- 1), and the respective cell parameters (A, °) [ a = 10.4427 (19), b = 12.881 (2), c = 15.991 (3), a= 76.840 (7), b= 85.130 (6), g= 87.278 (6), and [ a = 12.237 (3), b = 12.580 (3), c = 13.507 (3), a= 91.146 (8), b= 104.916 (8), g= 111.307 (8)]. Compound 3 crystallizes in the Monoclinic system ( P 2 1 / c) with the cell parameters (A, °) [a= 12.982 (3), b= 11.282 (2), c= 18.528 (4) and b (°) = 108.572 (7)]. The title compounds were evaluated for their biological activities against a range of cancer cell lines (BT-474, MDA-MB-231, AU565), Chronic myeloid leukemia cell line (K562), Lung cancer cell line (H460) and normal cell line 3T3 mouse fibroblast. Especially complexes 1 and 3 (derivatives of the piperic acid), i.e. Pipericcarboxylate triphenyltin [Ph 3 SnO 2 C-(CH=CH) 2 -C 7 H 5 O 2 ] n and {[n-Bu 2 SnO 2 C-(CH=CH) 2 -C 7 H 5 O 2 ] 2 O} 2 were most active against all cancer cell lines. These compounds were also active against 3T3 normal cell line, but the IC 50 values were high as compared to cancer cell lines.

Bovine HDL and Dual Domain HDL-Mimetic Peptides Inhibit Tumor Development in Mice.

A growing body of literature supports the role of apolipoproteins present in HDL in the treatment of pro-inflammatory diseases including cancer. We examined whether bovine HDL (bHDL) and three dual-domain peptides, namely AEM-28 and its analog AEM-28–2, and HM-10/10, affect tumor growth and development in mouse models of ovarian and colon cancer. We demonstrate that bHDL inhibits mouse colorectal cancer cell line CT26-mediated lung tumor development, and mouse ovarian cancer cell line ID8-mediated tumor burden. We also demonstrate that, although to different degrees, dual-domain peptides inhibit cell viability of mouse and human ovarian and colon cancer cell lines, but not that of normal human colonic epithelial cells or NIH3T3 mouse fibroblasts. Dual-domain peptides administered subcutaneously or in a chow diet decrease CT26 cell-mediated tumor burden, tumor growth, and tumor dissemination in BALB/c mice. Plasma levels of lysophosphatidic acid (LPA) are significantly reduced in mice that received bHDL and the dual-domain peptides, suggesting that reduction by effecting accumulation and/or synthesis of pro-inflammatory lipids may be one of the mechanisms for the inhibition of tumor development by bHDL and the dual-domain peptides. Our studies suggest that therapeutics based on apolipoproteins present in HDL may be novel agents for the treatment of epithelial adenocarcinomas of the ovary and colon.

Antileukemic Effect of Palladium Nanoparticles Mediated by White Tea (Camellia sinensis) Extract In Vitro and in WEHI‐3B‐Induced Leukemia In Vivo

This study investigated the in vivo antileukemic activity of palladium nanoparticles (Pd@W.tea-NPs) mediated by white tea extract in a murine model. The cell viability effect of Pd@W.tea-NPs, “blank” Pd nanoparticles, and white tea extract alone was determined in murine leukemia WEHI-3B cells and normal mouse fibroblasts (3T3 cells). Apoptotic and cell cycle arrest effects of Pd@W.tea-NPs in WEHI-3B cells were evaluated. The effects of Pd@W.tea-NPs administered orally to leukemic mice at 50 and 100 mg/kg daily over 28 days were evaluated. Pd@W.tea-NPs reduced the viability of WHEI-3B cells with IC50 7.55 μg/ml at 72 h. Blank Pd nanoparticles and white tea extract alone had smaller effects on WHEI-3B viability and on normal fibroblasts. Pd@W.tea-NPs increased the proportion of Annexin V-positive WHEI-3B cells and induced G2/M cell cycle arrest. Leukemic cells in the spleen were reduced by Pd@W.tea-NPs with an increase in Bax/Bcl-2 and cytochrome-C protein and mRNA levels indicating the activation of the mitochondrial apoptotic pathway. These effects replicated the effects of ATRA and were not observed using blank Pd nanoparticles. Pd@W.tea-NPs afford therapeutic efficacy against leukemia likely to pivot on activation of the mitochondrial pathway of apoptotic signaling and hence appear attractive potential candidates for development as a novel anticancer agent.

Therapeutic potential of xanthones from Swertia chirata in breast cancer cells.

Background & objectives:Medicinal plants like Swertia chirata are rich sources of different xanthones. This study was aimed to assess the cytotoxic potential of four most abundant xanthones present in S. chirata both in vivo and in vitro in Ehrlich ascites carcinoma (EAC), a mouse transplantable breast carcinoma cell line and two human breast carcinoma cell lines (MCF-7 and MDA-MB-231).Methods:Four xanthones derived from S. chirata namely 1-hydroxy-3,7,8-trimethoxyxanthone (XA), 1,8-dihydroxy-3,5-dimethoxyxanthone (XB), 1-hydroxy-3,5,8-trimethoxyxanthone (XC) and 1,5,8-trihydroxy-3-methoxyxanthone (XD) were used for determination of sub-lethal dose on the cell lines EAC, MCF-7, MDA-MB-231 and verified toxicity of sub-lethal dose on normal murine fibroblast cells. Cytotoxicity was measured in vivo and survivability of mice was plotted accordingly. Therapeutic efficacy of XD was evaluated both in vivo and in vitro by determination of lipid peroxidation (LPO), reactive oxygen species (ROS) generation and by quantitating the enzyme status (GSH, catalase, superoxide dismutase) in treated and untreated samples. DNA damage was evaluated using comet and DNA fragmentation assays. Furthermore, apoptotic effect was analyzed by flow cytometry and validated by TUNEL assay and Western blotting.Results:Among all the xanthones tested XD showed IC50 at the lowest dose, and normal cells were unaffected at this dose. Survivability of mice increased significantly when treated with XD compared to other xanthones and cisplatin. Significantly increased ROS and LPO were found in cancer cells as a result of XD treatment which was unaltered in normal cell line. XD induced DNA damage and apoptosis in cancer cell lines.Interpretation & conclusions:Our experimental data indicate that XD may potentially act as a chemotherapeutic agent by enhancing ROS in breast cancer cells thereby leading to apoptosis.

The terminal sialic acid of stage-specific embryonic antigen-4 has a crucial role in binding to a cancer-targeting antibody

The terminal sialic acid of stage-specific embryonic antigen-4 has a crucial role in binding to a cancer-targeting antibody

Comparison of cytotoxicity and smear layer removal efficacy of triphala (an Indian ayurvedic herbal formulation) and 5.25% sodium hypochlorite as root canal irrigants: An in vitro study.

Healing potential of plants is an age-old idea that has recently attained renewed interest. Considering the ineffectiveness, potentially harmful effects, and safety concerns of commonly used synthetic irrigants, the herbal alternatives for endodontic usage might prove to be advantageous. The aim of this study was to evaluate the adequacy of smear layer removal and cytotoxicity potential of triphala in comparison to sodium hypochlorite. The study was conducted in two parts: the first part of the study was cytotoxicity assessment studied using Alamar blue assay. L929 mouse fibroblasts were seeded in 96-well plates at a density of 5000 cells/well and treated with different concentrations of triphala and NaOCl for a period of 24 and 48 h. The percentage of cell viability was then quantified using an Alamar blue assay. The optical density was measured at 570 nm and compared with 620 nm, which was considered as a reference wavelength. The second part of the study was smear layer assessment at the coronal, middle, and apical third of twenty human premolar teeth using scanning electron microscope. The Alamar blue reagent cytotoxicity study suggested that triphala showed no cytotoxic properties against the normal mouse fibroblast cells whereas sodium hypochlorite showed a significant cytotoxic effect against the L929 cell lines with the IC50 concentration at 1.8%, respectively, after the treatment of 24 h of incubation at 37°C temperature. Triphala was as effective as sodium hypochlorite in smear layer removal in the coronal and middle third of the root, but sodium hypochlorite showed better smear layer removal in the apical third. Triphala can be considered as a superior irrigant with good antibacterial efficacy and least cytotoxicity potential compared to conventional hypochlorite irrigating agent and provide adequate clearing of smear layer in the coronal and middle third, and further studies are warranted to alter the properties of liquid to make it more cleansable in the apical third of the root.

Comparative Study of Cytotoxic Activity of Nano Silver Against A549 and L929 Cell Lines

Studies in recent years are focussed on anticancer drugs which can selectively induce cell death with less toxicity to normal cells. The present work therefore aims at exploring the potential of nano silver as selective anticancer drug by comparing its cytotoxic activity against human lung carcinoma cell line (A549) and mouse normal fibroblast cell line (L929) in vitro. Nano silver was synthesized by both chemogenic (AgNP-C) and biogenic (AgNP-B) method and characterized by using PXRD, SEM and TEM. In order to assess the molecular mechanism involved in cytotoxicity, apoptosis inducing effect of nano silver was assessed by Annexin V/PI staining, cell cycle analysis and caspase-3 expression study. From the results, it was confirmed that A549 cells treated with nano silver showed decreased cell viability (AgNP-C: 173.5 ± 2.51 μg/mL, AgNP-B: 29.2 ± 0.22 μg/mL) compared to L929 cells (AgNP-C: 317.2 ± 3.43 μg/mL, AgNP-B: 622.3 ± 1.6 μg/mL), indicating lower toxicity of nano silver towards normal cells. Apoptotic study, cell cycle analysis and caspase-3 studies showed decreased expression of Bcl-2 and increased expression of Bax mitochondrial genes facilitating release of cytochrome c (cyt c) into cytosol by disrupting mitochondrial membrane potential indicating induction of cell death in A549 cells through mitochondrial mediated intrinsic apoptosis pathway. Present investigation provides conclusive evidence for application of biogenic nano silver as a potential candidate for anticancer drug development.

Sivas İlinde Doğal Olarak Yetişen ve Yenilebilir Bir Bitki Olan Evelik (Rumex crispus) Bitkisinin Biyoaktivitesi ve Terapotik Nitelikleri

Bitkilerden elde edilen ekstrelerin birçoğunun biyolojik etkileri ve etki mekanizmaları hakkındaki bilimsel veriler hala yetersiz olmakla birlikte bu konuya olan ilgi her geçen gün artmaktadır. Bu çalışmada, Rumex crispus (Evelik) bitkisi mevsiminde Sivas yöresinden toplanarak kurutulmuş ve sonrasında yapraklı kısımlarından etanol, metanol ve su ekstraksiyonları elde edilmiştir. Çalışmanın amacı, elde edilen her bir ekstrenin bazı mikroorganizmalar üzerindeki antimikrobiyal aktivitelerinin değerlendirilmesi, toplam antioksidan seviyesi (TAS), toplam oksidan seviyesi (TOS), oksidatif stres indeksi (OSI) değerlerinin ve çeşitli hücre hatları üzerindeki anti-kanserojen niteliklerinin ortaya konulmasıdır. TAS, TOS ve OSI değerlerini tespit etmek için Rel Assay Diagnostics kitler kullanılmıştır. Bitki ekstrelerinin mikroorganizmalara karşı minimum inhibisyon konsantrasyonunu (MIC) belirlemek amacıyla mikrodilüsyon broth yöntemi kullanılmıştır. Çalışmada antimikrobiyal aktivite analizleri, Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), Bacillus cereus (ATCC 11778), Klebsiella pneumonia (ATCC 13883), Candida albicans (ATCC 10231) ve Candida tropicalis (DSM 11953) mikroorganizmaları kullanılarak yapılmıştır. Normal fare fibroblast hücre hattı ve 3 farklı insan kanser hücre hattında MTT yöntemi ile bitki özütlerinin sitotoksik aktivitesi belirlenmiştir. Sonuç olarak, evelik bitkisinin etanol, metanol ve su ekstraklarının denendiği mikroorganizmalar üzerinde antimikrobiyal aktivitesinin bulunmadığı, yüksek düzeyde antioksidan aktiviteye sahip olduğu ve bazı hücre kültürlerinde sitotoksik aktivitesinin olduğu tespit edilmiştir.

Sivas İlinde Kültürü Yapılan Kenger (Gundellia tournefortii L.) Bitkisi Tohumlarının Besin Elementlerinin ve Biyolojik Aktivitelerinin Değerlendirilmesi

Bu çalışmada, Gundellia tournefortii bitkisinin tohumlarından elde edilen su ekstraktının GC/MS (Gaz kromatografisi -Kütle spektrometresi) analizi ile genel olarak içeriğinin değerlendirilmesi, makro ve mikro besin elementi konsantrasyonları, antimikrobiyal aktivitesi, toplam antioksidan seviyesi (TAS), toplam oksidan seviyesi (TOS), oksidatif stres indeksi (OSI) değerlerinin belirlenmesi ve çeşitli hücre hatları üzerindeki anti-kanserojen niteliklerinin ortaya konulması amaçlanmıştır. TAS, TOS ve OSI değerlerini tespit etmek için Rel Assay Diagnostics kitler kullanılmıştır. Antimikrobiyal aktivite analizleri, Mikrodilüsyon Broth yöntemi ile Minimum inhibisyon konsantrasyon (MIC) değeri belirlenerek, Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), Bacillus cereus (ATCC11778), Klebsiella pneumonia (ATCC 13883), Candida albicans (ATCC 10231) ve Candida tropicalis (DSM11953) mikroorganizmaları üzerinde yapılmıştır. Sitotoksik aktivite ise, normal fare fibroblast hücre hattı, HUVEC ve 3 farklı insan kanser hücre hattında MTT yöntemi ile belirlenmiştir. Sonuç olarak, Gundelia tournefortii bitki ekstraktının S. aureus bakterisi üzerinde orta düzeyde, diğer mikroorganizmalar üzerinde ise zayıf düzeyde antimikrobiyal aktivite, iyi derecede antioksidan aktivite ve bazı hücrelerde sitotoksik aktivitesinin olduğu tespit edilmiştir. Bitkinin oksidatif stres indeksinin ise, düşük olduğu belirlenmiştir. Ayrıca, makro ve mikro besin elementi içeriği bakımından Gundellia tournefortii bitkisinin %3,64 N, %0,11 P, %3,78 K, %0,22 Ca, %0,57 Mg, 268,4 mg/kg Fe, 16,7 mg/kg Zn, 19,4 mg/kg Mn ve 8,3 mg/kg Cu konsantrasyonlarına sahip olduğu belirlenmiştir.

Formononetin induces apoptotic cell death through the suppression of mitogen‑activated protein kinase and nuclear factor‑κB phosphorylation in FaDu human head and neck squamous cell carcinoma cells.

Formononetin, a phytoestrogen extracted from various herbal plants, has been investigated as an anticancer agent against diverse types of cancer. The aim of the present study was to investigate the induction of apoptotic cell death by formononetin in the FaDu pharyngeal squamous cell carcinoma cell line. Formononetin significantly increased FaDu cell death, with an estimated IC50 value of 50µM; however, it did not affect the viability of normal L929 mouse fibroblasts used as normal control at 5‑25µM. Typical characteristics of apoptosis, such as morphological alterations, chromatin condensation, DNA fragmentation and the size of the apoptotic cell population, were increased in FaDu cells treated with formononetin for 24h. Furthermore, formononetin‑induced FaDu cell death involved the death receptor‑mediated extrinsic and the mitochondria‑dependent intrinsic apoptotic pathways by activating the caspase cascade. The chemotherapeutic effects of formononetin were mediated by the suppression of mitogen‑activated protein kinases, including extracellular signal‑regulated kinase1/2 and p38, and nuclear factor‑κB phosphorylation in FaDu cells. Finally, the oral administration of formononetin decelerated tumor growth through the expression of cleaved caspase‑3 in a FaDu cell xenograft animal model. Taken together, these findings indicate that formononetin holds promise as a chemotherapeutic agent and may be of value in the treatment of human head and neck squamous cell carcinoma.