Healthy Fibroblast Cell Line Research Articles

Effective Radiosensitization of HNSCC Cell Lines by DNA-PKcs Inhibitor AZD7648 and PARP Inhibitors Talazoparib and Niraparib.

(1) Head and neck squamous cell carcinoma (HNSCC) is common, while treatment is difficult, and mortality is high. Kinase inhibitors are promising to enhance the effects of radiotherapy. We compared the effects of the PARP inhibitors talazoparib and niraparib and that of the DNA-PKcs inhibitor AZD7648, combined with ionizing radiation. (2) Seven HNSCC cell lines, including Cal33, CLS-354, Detroit 562, HSC4, RPMI2650 (HPV-negative), UD-SCC-2 and UM-SCC-47 (HPV-positive), and two healthy fibroblast cell lines, SBLF8 and SBLF9, were studied. Flow cytometry was used to analyze apoptosis and necrosis induction (AnnexinV/7AAD) and cell cycle distribution (Hoechst). Cell inactivation was studied by the colony-forming assay. (3) AZD7648 had the strongest effects, radiosensitizing all HNSCC cell lines, almost always in a supra-additive manner. Talazoparib and niraparib were effective in both HPV-positive cell lines but only consistently in one and two HPV-negative cell lines, respectively. Healthy fibroblasts were not affected by any combined treatment in apoptosis and necrosis induction or G2/M-phase arrest. AZD7648 alone was not toxic to healthy fibroblasts, while the combination with ionizing radiation reduced clonogenicity. (4) In conclusion, talazoparib, niraparib and, most potently, AZD7648 could improve radiation therapy in HNSCC. Healthy fibroblasts tolerated AZD7648 alone extremely well, but irradiation-induced effects might occur. Our results justify in vivo studies.

Biological Evaluation of Azide Derivative as Antibacterial and Anticancer Agents

Azides are significant compounds because of their biological effects on bacteria, viruses, fungi, and cancer. The antibacterial and anticancer properties of azide derivatives have been demonstrated in several microbial strains and cancer cell lines. The most significant microorganisms in command of hospital-acquired nosocomial infections include Acinetobacter baumannii (A. baumannii), Methicillin-resistant Staphylococcus aureus (MRSA), Candida albicans (C. albicans), and Candida dubliniensis (C. dubliniensis). These infections have become resistant to a variety of antibiotics. These resistant infections are frequently seen in cancer patients. Novel drug-active compounds are thus required that have both antibacterial and anticancer activity. This study examined previously synthesized 1-azido-5,6,7-trimethoxy-2,3-dihydro-1H-indene (4) for its antibacterial and anticancer activities. Antimicrobial activity was assessed using the disc diffusion technique, and minimum inhibitory concentration values were computed for zone formation in the examined pathogens. The study's findings indicate that while azide derivatives did not exhibit any effect against other pathogens, they suppressed bacterial growth with a zone diameter of 10 mm against A. baumannii. The MIC values of azide derivative against A. baumannii were 3.90 µg/ml. WST-8 analysis in Caco-2 cancer and healthy fibroblast cell line was used to determine the anticancer study. As a result of this analysis, the IC50 value was calculated to be 2.99 µM. There is little anticancer activity. A very low toxic effect of the azide compound on fibroblast cells was also observed. These findings suggest that this azide derivative may be tested as a potential antibacterial and anticancer agent.

ARCTİUM MİNUS METANOL EKSTRESİNİN ÇEŞİTLİ KANSER HÜCRE HATLARI ÜZERİNDE SİTOTOKSİK ETKİLERİ

Objective: This study aimed to evaluate the cytotoxic effects of Arctium minus (Hill) Bernh. ssp. minus methanol extract derived from aerial parts on cancer cell lines.Material and Method: For cytotoxicity assays, two different human breast cancer cell lines (MCF-7 and MDA-MB-231) and healthy human fibroblast cell line (MRC-5)as a control were used. Cell viability determination was performed using the CellTiter-Blue method. One-Way ANOVA and Tukey post test were used for statistical analysis.Result and Discussion: Cell viability has been detected between ratios of 27.8-38.7% for MCF-7 cancer cell line, and a significant cytotoxic activity was observed via the analysis (1 mg/mL extract treatment p< 0.022). However, 47.8-59.7% cell viability was observed for MDA-MB-231 cancer cell line, and MRC-5 healthy fibroblast cell line did not demonstrate any cell viability (92.4-105.4% cell viability). Depending on these data, MCF-7 cancer cell line and MRC-5 fibroblast healthy cell line were treated with Arcitum minus extract, then cell viability was detected by flow cytometry technique. The ratio of the cell death was higher in MCF-7 cancer cell line (98%) compared with the MRC-5 fibroblast healthy cell line (25%) after the Arctium minus extract treatment. In conclusion, Arctium minus ssp. minus extract has significantly decreased the cell viability in MCF-7 cancer cell line when compared with the MCR-5 fibroblast normal cell line.

Effect of Wheat Germ Oil on Wound Healing: An In Vitro Study in Fibroblast Cells

Purpose: Wound healing is a set of mechanisms that are activated to restore structurally damaged tissue. There are many studies aimed at accelerating wound healing. In this context, products obtained from plants come to the fore. In this article, the effects of wheat germ oil (WGO), which is known to have antioxidant and anti-inflammatory properties, on wound healing were investigated by in vitro method.
 Material and Methods: Wheat germ oil and α-tocopherol were applied to L929, a healthy fibroblast cell line, at different doses for 24, 48 and 72 hours. Cell viability was measured by XTT colorimetric method. In vitro wound healing model was applied at the dose where the oil was effective. Obtained results were analyzed statistically.
 Results: As a result of the application of α-tocopherol to L929 cells at different doses, it was observed that there was no significant contribution to cell proliferation compared to the control group. However, WGO was observed to significantly increase proliferation at the 100 ng/ml concentration. In the wound healing model, cells treated with WGO at 48 hours were observed to proliferate faster and invade the wound site more rapidly. (p

Molecular docking and in vitro anticancer studies of silver(I)-N-heterocyclic carbene complexes

A series of symmetric and unsymmetrical benzimidazolium-based N-heterocyclic carbene (NHC) precursors (1a-i) and their silver complexes (2a-i) have been synthesized. The Ag(I)–NHC complexes were characterized by 1H, 13C{1H} NMR, FTIR, LC/MS-QTOF, and elemental analysis. Anticancer and cytotoxic activity of all Ag(I)–NHC complexes were tested against healthy fibroblast cell line (L929), breast cancer cell line (MCF-7), and neuroblastoma cell line (SH-SY5Y) by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4sulfophenyl)-2H-tetrazolium] assay. The 2b, 2c, 2e, 2g, 2h, and 2i complexes showed higher cytotoxicity than cisplatin against SH-SY5Y and MCF-7 and lower cytotoxic activity against L929 cell lines. Because of their high cytotoxic activity against cancer cells and low cytotoxicity against healthy fibroblast cell lines, the 2b, 2c, 2e, 2g, 2h, and 2i are expected to be new lead compounds. In addition, molecular docking studies were performed to explore the binding interactions of silver complexes with the enzyme to explore new anticancer compounds. Furthermore, ADME properties of all complexes were predicted to explore lead-like characteristics and may be a potential drug candidate for cancer treatment.

Synthesis of diaryl urea derivatives and evaluation of their antiproliferative activities in colon adenocarcinoma

Colon cancer is one of the leading causes of cancer-related deaths today. Serious research for ideal chemotherapy continues today. In this context, newly synthesized molecules have an essential role in cancer treatment research. The effects of 5 diaryl urea derivatives synthesized within the scope of this study on the HT-29 colon cancer cell line were investigated for the first time in the literature by in-silico and in-vitro methods. Among the five compounds produced in the first stage of the study, DAU5 was found to have a cytotoxic effect on HT-29. However, it was determined that it did not show a serious cytotoxic effect on L929 (healthy fibroblast cell line) at the same doses. The efficacy of DAU5 was evaluated for expression of LC3B, an indicator of autophagy, and 8-OHdG, an indicator of oxidative stress-DNA damage. LC3B and 8-OHdG expression were lower in the DAU5 treatment group than in the control group. As a result, it was seen that DAU5, a diaryl urea derivative compound we synthesized in this study, has the potential to be a chemotherapeutic agent against colon cancer. However, our study needs to be supported by in vivo and clinical studies.

MitoCellPhe reveals mitochondrial morphologies in single fibroblasts and clustered stem cells.

Mitochondria are dynamic organelles that differ significantly in their morphologies across cell types, reflecting specific cellular needs and stages in development. Despite the wide biological significance in disease and in health, delineating mitochondrial morphologies in complex systems remains challenging. Here, we present the Mitochondrial Cellular Phenotype (MitoCellPhe) tool developed for quantifying mitochondrial morphologies and demonstrate its utility in delineating differences in mitochondrial morphologies in a human fibroblast and human induced pluripotent stem cell (hiPSC) line. MitoCellPhe generates 24 parameters, allowing for a comprehensive analysis of mitochondrial structures and importantly allows for quantification to be performed on mitochondria in images containing single cells or clusters of cells. With this tool, we were able to validate previous findings that show networks of mitochondria in healthy fibroblast cell lines and a more fragmented morphology in hiPSCs. Using images generated from control and diseased fibroblasts and hiPSCs, we also demonstrate the efficacy of the toolset in delineating differences in morphologies between healthy and the diseased state in both stem cell (hiPSC) and differentiated fibroblast cells. Our results demonstrate that MitoCellPhe enables high-throughput, sensitive, detailed, and quantitative mitochondrial morphological assessment and thus enables better biological insights into mitochondrial dynamics in health and disease.

Theranostic Mesoporous Silica Nanoparticles Loaded With a Curcumin-Naphthoquinone Conjugate for Potential Cancer Intervention.

A novel theranostic molecule, derived from curcumin (Cur) and naphthoquinone (NQ), allowing for cancer targeting, detection and treatment was previously described and termed CurNQ. To allow for enhanced theranostic capabilities, advanced drug delivery techniques are required. To this end, mesoporous silica nanoparticles (MSN) were synthesized and CurNQ was loaded into its pores to form the novel nanosystem MSN_CurNQ. The formation of the nanosystem aimed to augment the drug delivery of CurNQ through the EPR effect and sustained release. Moreover, the loading of CurNQ into its pores, formed a fluorescent nanoparticle that can be tracked, detected and visualized. Herein, the synthesis of a novel nanosystem is described and its theranostic potential are explored in vitro. MSN with an average size of 108 d.nm, a zeta potential of −42 mV and a PDI of 0.150 were synthesized and were impregnated with CurNQ to form the novel nanosystem MSN_CurNQ. MSN_CurNQ was demonstrated to have pH-responsivity whereby after 96 h, at pH 7.4, 31.5% of CurNQ was released from the MSN compared to 57% release at pH 6.8, corresponding to an increase of 25.5% in release with a 0.6 pH drop. The innate fluorescence was then characterized through confocal and fluorescence microscopy. Microscopy images illustrated the distinct, high intensity innate fluorescence with a high background to target ratio, thus confirming detection capabilities and potentially extending MSN_CurNQ’s application to molecular imaging purposes. Moreover, the chemotherapeutic potential of MSN_CurNQ was demonstrated as cell viability was reduced to below 50% in OVCAR-5, CACO-2, CHLA, and MCF-7 cell lines. Furthermore, MSN_CurNQ displayed tumor specific toxicity whereby the cell viability was reduced to a far greater extent in the cancer cell lines compared to a healthy fibroblast cell line (p = 0.000). Indeed, the novel MSN_CurNQ nanosystem has potential for applications in cancer targeting, detection and treatment.

Synthesis and Biological Screening of New 4-Hydroxycoumarin Derivatives and Their Palladium(II) Complexes.

Two newly synthesized 4-hydroxycoumarin bidentate ligands (L1 and L2) and their palladium(II) complexes (C1 and C2) were screened for their biological activities, in vitro and in vivo. Structures of new compounds were established based on elemental analysis, 1H NMR, 13C NMR, and IR spectroscopic techniques. The obtained compounds were tested for their antioxidative and cytotoxic activities and results pointed to selective antiradical activity of palladium(II) complexes towards •OH and -•OOH radicals and anti-ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical) activity comparable to that of ascorbate. Results indicated the effect of C1 and C2 on the enzymatic activity of the antioxidative defense system. In vitro cytotoxicity assay performed on different carcinoma cell lines (HCT166, A375, and MIA PaCa-2), and one healthy fibroblast cell line (MRC-5) showed a cytotoxic effect of both C1 and C2, expressed as a decrease in carcinoma cells' viability, mostly by induction of apoptosis. In vivo toxicity tests performed on zebrafish embryos indicated different effects of C1 and C2, ranging from adverse developmental effect to no toxicity, depending on tested concentration. According to docking studies, both complexes (C1 and C2) showed better inhibitory activity in comparison to other palladium(II) complexes.

Improved Antiproliferative Activity and Fluorescence of a Dinuclear Gold(I) Bisimidazolylidene Complex via Anthracene-Modification.

A straightforward modification route to obtain mono‐ and di‐substituted anthroyl ester bridge functionalized dinuclear Au(I) bis‐N‐heterocyclic carbene complexes is presented. The functionalization can be achieved starting from a hydroxyl‐functionalized ligand precursor followed by transmetallation of the corresponding Ag complex or via esterification of the hydroxyl‐functionalized gold complex. The compounds are characterized by NMR‐spectroscopy, ESI‐MS, elemental analysis and SC‐XRD. The mono‐ester Au complex shows quantum yields around 18%. In contrast, the corresponding syn‐di‐ester Au complex, exhibits significantly lower quantum yields of around 8%. Due to insufficient water solubility of the di‐ester, only the mono‐ester complex has been tested regarding its antiproliferative activity against HeLa‐ (cervix) and MCF‐7‐ (breast) cancer cell lines and a healthy fibroblast cell line (V79). IC50 values of 7.26 μM in the HeLa cell line and 7.92 μM in the MCF‐7 cell line along with selectivity indices of 8.8 (HeLa) and 8.0 (MCF‐7) are obtained. These selectivity indices are significantly higher than those obtained for the reference drugs cisplatin or auranofin.

Investigation of the Anticancer and Proliferative Effect of Broccoli Extract on Du145 Prostate Cancer and MEF Healthy Fibroblast Cell Lines

In recent years, many researchers have focused on the health effects of plant-derived foods. In this context, foods with high content of flavonoids and phenolic substances have received a great deal of attention as potential agents for cancer prevention and treatment. Studies on the broccoli plant have revealed that broccoli has antioxidant and anti-carcinogenic properties. However, effective dose of broccoli needs to be determined for the cancer treatment. In this study, the effects of broccoli extracts on du145 prostate cancer and Mef fibroblast healthy cells were investigated at different doses for 48 hours. Du145 and Mef cells were grown with Dulbecco's Minimum Essential Medium (DMEM) and HAMS F 12 (1: 1) supplemented with 2% FBS. Broccoli extracts at ten different doses (0,19% - 100%) were added into cultures and incubated at 37°C for 24 and 48 h in 5% CO2. The viability of the cells was determined by the MTT method (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide). Probit analysis by SPSS software revealed that the broccoli extract at a dose of 4.070 % dose killed du145 cancer cells at 48 h. At the same time, MTT results showed that the viability of Mef cells was increased during 48 h of incubation. As a result, broccoli extract showed a significant level of anticancer activity in Du145 cells, while increasing the viability of Mef healthy cells. This result suggests that broccoli extract is a potential candidate for cancer treatment.

Potential Therapeutic Activity of Bio-Synthesized Silver Nanoparticles as Anticancer and Antimicrobial Agent

Aloevera is one of the oldest medicinal plants and it possesses different types of pharmacological properties including anti-microbial, anti-inflammatory, anti-cancer and antioxidant activity. More over Aloevera contains 75 bioactive compounds such as polyphenol, flavonoids, alkaloids, anthraquinone etc. Such bioactive compounds can be used to reduce silver ions to produce silver nanoparticles. In this study, we are trying to formulate bio-synthesised silver nanoparticle (b-AgNps) by green chemistry approach. It is a very simple, efficient and eco-friendly approach for silver nanoparticle synthesis that is formed by reduction of silver nitrate (AgNO3) solution using Aloe vera leaf extract as a reducing agent.Hydrothermal method was used to prepare b-AgNps using aloe vera leaf extract as both reducing and stabilizing agent. Then b-AgNps were characterized by FESEM, XRD, DLS and UV-VIS spectroscopy. Biological activity of b-AgNps were evaluated by performing cytotoxic test (MTT assay) against breast cancer cell lines and Fibroblast normal cell lines as well as screening antimicrobial activity using agar well diffusion method. The results revealed that there was significant amount of cell growth inhibition against breast cancer cell lines as well as microbes compare to normal healthy fibroblast cell lines.

Real-time Analysis of Impedance Alterations by the Effects of Vanadium Pentoxide on Several Carcinoma Cell Lines.

Vanadium compounds have various pharmacologic effects and all available evidence reveals that the effects of vanadium compounds depend on many factors, mainly on the type of cells and dose. The proapoptotic or antiapoptotic effect of vanadium compounds depends strongly on the cell type. In this study, the effects of vanadium pentoxide (V2O5) were investigated using several tumor cell lines: a colorectal cancer cell line (Colo-205), a human breast adenocarcinoma cell line (MCF-7), and a normal human fibroblast cell line. Five different concentrations of V2O5 between 25-200 µM were applied on the cells and xCELLigence real-time cell analysis was conducted to evaluate the impedance alterations. This study is the first to show V2O5's effects on Colo-205 and MCF-7 and human fibroblast cell lines in a real-time manner. In the Colo-205 cell line, cell index (CI) alterations decreased slightly at 25 µM and 50 µM, and increased at 100 µM, 150 µM and 200 µM concentrations. In the MCF-7 cell line, CI alterations increased at all concentrations compared with the untreated control. However, in the healthy fibroblast cell line, the CI alterations decreased at all concentrations compared with the untreated control, which limits the use of V2O5 for its cytotoxic effect in vivo. The combination of conventional anticancer drugs can be used to increase the effectiveness and reduce the adverse effects of these drugs considering stages of cancer and cancer type. Our results suggest that V2O5 has disparate effects on several cancer cells at different concentrations.

Cold storage of biopsies from wild endangered native Chilean species in field conditions and subsequent isolation of primary culture cell lines

We evaluated the possibility of deriving primary cell cultures from tissue biopsies taken in field conditions from six threaten endemic Chilean species of free-ranging mammals. Biopsies were taken either by ear punching or darts fired to animals and hold in hypothermic conditions (4 degrees C) in defined salt solution for time periods ranging from 0 to 7 d before biopsy samples reached the cell culture laboratory. Previously, holding times were evaluated in experimental cows in controlled conditions. Two enzymatic treatments, collagenase alone or collagenase followed by tripsin, were used to disaggregate tissues for cell culture. We found that ear notches and dart-derived biopsies can be storaged at 4 degrees C for 1 wk and still yield primary cultures. For dart-derived biopsies, there was an invert correlation between length of cold storage and cellular viability in culture. Healthy fibroblast cell lines were obtained in 92% of the biopsies taken despite the origin (punch or biopsy). We are not aware of similar study for free-ranging animals, especially for the use of darting system to biopsy wild terrestrial mammals, we believe that our results could help for a more widespread implementation of these procedures in the practice of ex situ conservation.