scholarly journals Cell Electropermeabilisation Enhancement by Non-Thermal-Plasma-Treated PBS

Cancers ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 219 ◽  
Author(s):  
Thai-Hoa Chung ◽  
Augusto Stancampiano ◽  
Kyriakos Sklias ◽  
Kristaq Gazeli ◽  
Franck André ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Electric Fields ◽  
Combined Treatment ◽  
Chinese Hamster ◽  
Field Amplitude ◽  
Lung Fibroblasts ◽  
Cancer Therapies ◽  
Electric Pulses ◽  
Combined Application

The effectiveness of electrochemotherapy (ECT) in local eradication of tumours in human and veterinary medicine has been proven. ECT consists of increasing the uptake of cytotoxic drugs by means of pulsed electric fields (PEFs) that transiently permeabilise the cell membrane. Still, this tumour treatment includes some drawbacks that are linked to the characteristics of the intense electric pulses (EPs) used. Meanwhile, the emerging field of cancer therapies that are based on the application of non-thermal plasmas (NTP) has recently garnered interest because of their potentialities as rich sources of reactive species. In this work, we investigated the potential capabilities of the combined application of indirect NTP treatment and microsecond PEFs (µsPEFs) to outperform in vitro cell electropermeabilisation, the basis of ECT. Thus, phosphate-buffered saline (PBS) was plasma-treated (pPBS) and used afterwards to explore the effects of its combination with µsPEFs. Analysis of two different cell lines (DC-3F Chinese hamster lung fibroblasts and malignant B16-F10 murine melanoma cells), by flow cytometry, revealed that this combination resulted in significant increases of the level of cell membrane electropermeabilisation, even at very low electric field amplitude. The B16-F10 cells were more sensitive to the combined treatment than DC-3F cells. Importantly, the percentage of permeabilised cells reached values similar to those of cells exposed to classical electroporation field amplitude (1100 V/cm) when the cells were treated with pPBS before and after being exposed only to very low PEF amplitude (600 V/cm). Although the level of permeabilisation of the cells that are treated by the pPBS and the PEFs at 600 V/cm is lower than the level reached after the exposure to µsPEFs alone at 1100 V/cm, the combined treatment opens the possibility to reduce the amplitude of the EPs used in ECT, potentially allowing for a novel ECT with reduced side-effects.

8. Evaluation of Snow Fungus Polysaccharides on benzo[a]pyrene-induced DNA Damage

2018 ◽  
Author(s):  
Daisy Liu
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Free Radical ◽  
Radical Scavenging ◽  
Chinese Hamster ◽  
Negative Control ◽  
Lung Fibroblasts ◽  
Protective Effects ◽  
Tremella Fuciformis

Snow fungus, Tremella fuciformis, has been demonstrated to have numerous health benefits including purported chemopreventive properties due to free radical-scavenging ability. Protective effects derived from snow fungus polysaccharides are evaluated on Chinese hamster lung fibroblasts (CCL-39) exposed to carcinogen benzo[a]pyrene known to cause free radical formation and oxidative stress to cells. In this experiment, it was hypothesized that the naturally occurring polysaccharides in snow fungus are able to protect against or reduce oxidative stress-induced DNA damage. Polysaccharides were isolated through an alkaline extraction and in-vitro digestion. DNA damage was measured using the single-cell gel electrophoresis comet assay after exposure to benzo[a]pyrene and polysaccharide extract to lung fibroblasts. Results were calculated using the mean and standard deviation data of tail length and area, respectively. Each damaged cell was measured and analyzed through ImageJ Editing Software. The results indicate a promising trend which depict snow fungus polysaccharides yielding lower levels of DNA damage compared to cells exposed to benzo[a]pyrene and compared to the negative control (phosphate buffered saline and Dulbecco’s cell medium). This study suggests polysaccharides from Tremella fuciformis could truly prevent cellular DNA damage by protecting against oxidative stress.

The Attachment of V79 and Human Periodontal Ligament Fibroblasts on Periodontally Involved Root Surfaces Following Treatment with EDTA, Citric Acid, or Tetracycline HCL: An SEM in vitro Study

2006 ◽  
Vol 7 (1) ◽  
pp. 35-43 ◽  
Author(s):  
R. Viswa Chandra ◽  
Ganesh Chandra Jagetia ◽  
K. Mahalinga Bhat
Keyword(s):  
Citric Acid ◽  
Periodontal Ligament ◽  
Chinese Hamster ◽  
Lung Fibroblasts ◽  
Control Group ◽  
Periodontal Ligament Fibroblasts ◽  
Human Periodontal Ligament Fibroblasts ◽  
Root Surfaces ◽  
Tetracycline Hcl

Abstract Objective The present in vitro study has been designed to establish and compare the effects of citric acid, EDTA, and tetracycline HCl on human periodontally diseased roots on the structure, attachment, and orientation of V79 (primary Chinese hamster lung fibroblasts) cells and human periodontal ligament fibroblasts (HPDL). Materials and Methods Commercially availableV79 cells and HPDL derived from healthy human third molars were used in this study. These fibroblasts were left in solution for seven days in order to attain confluence. Forty single-rooted teeth were obtained from patients diagnosed with periodontitis. The crown part was removed under constant irrigation and the root was split vertically into two equal halves, thus, yielding 80 specimens. Following scaling and root planing, the specimens were washed with phosphate buffered saline (PBS) and kept in 50 μg/ml gentamycin sulphate solution for 24 hours. The root pieces were then treated as follows: citric acid at pH 1, 24% EDTA, or with a 10% solution of tetracycline HCl and were then placed in V79 fibroblast cultures and HPDL cultures. The specimens were harvested after four weeks and were fixed in 2.5% glutaraldehyde in PBS before preparation for scanning electron microscopy (SEM). Results The behavior of V79 cells was similar to that of human periodontal ligament cells on root conditioned surfaces. V79 and HPDL showed a healthy morphology on root surfaces treated with citric acid and EDTA and a relatively unhealthy appearance on root surfaces treated with tetracycline HCl and distilled water (control group). Conclusion The results suggest the use of citric acid and EDTA as root conditioning agents favorably affects the migration, attachment, and morphology of fibroblasts on human root surfaces, which may play a significant role in periodontal healing and regeneration. Citation Chandra RV, Jagetia GC, Bhat KM. The Attachment of V79 and Human Periodontal Ligament Fibroblasts on Periodontally Involved Root Surfaces Following Treatment with EDTA, Citric Acid, or Tetracycline HCL: An SEM in vitro Study. J Contemp Dent Pract 2006 February;(7)1:044-059.

2-Amino-3-methylimidazo[4, 5-f]quinoline (IQ), a carcinogenic pyrolysate, induces chromosomal aberrations in Chinese hamster lung fibroblasts in vitro

Mutagenesis ◽  
1993 ◽  
Vol 8 (4) ◽  
pp. 349-354 ◽  
Author(s):  
Kunihiko F. Miura ◽  
Midori Hatanaka ◽  
Chino Otsuka ◽  
Takatomo Satoh ◽  
Hiroko Takahashi ◽  
...  
Keyword(s):  
Chromosomal Aberrations ◽  
Chinese Hamster ◽  
Lung Fibroblasts

scholarly journals Electroporation with Cisplatin against Metastatic Pancreatic Cancer:In VitroStudy on Human Primary Cell Culture

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Olga Michel ◽  
Julita Kulbacka ◽  
Jolanta Saczko ◽  
Justyna Mączyńska ◽  
Piotr Błasiak ◽  
...  
Keyword(s):  
Primary Culture ◽  
Cell Lines ◽  
Lung Metastases ◽  
Cellular Membrane ◽  
Ductal Adenocarcinoma ◽  
Rapid Progression ◽  
Cancer Therapies ◽  
Electric Pulses ◽  
Pancreatic Cancer Cells

Despite the rapid progression of cancer pharmacotherapy, the high drug resistance of pancreatic ductal adenocarcinoma (PDA) makes it one of the most lethal malignancies. Therefore, there are high expectations associated with experimental therapies, such as electrochemotherapy (ECT). This technique involves the application of short electric pulses to induce transitional permeabilization of the cellular membrane, thus enhancing drug molecules influx. The aim of the study was to investigate the influence of electroporation with cisplatin (CisEP) on the primary culture of human PDA cells from lung metastases—their survival and stress response. Considering the growing importance of various research models, two established human PDA cell lines, EPP85-181P (sensitive to daunorubicin) and EPP85-181RDB (resistant to daunorubicin), were utilized as a reference control. Cisplatin revealed higher cytotoxicity towards established cell lines. Following CisEP application, we observed a significant decrease of cells viability in the primary culture model. After CisEP therapy, an increased immunoreactivity with SOD-2 and Casp-3 antibodies was noticed. In conclusion, we discovered that electroporation can enhance the cytotoxic effect of cisplatin in pancreatic cancer cellsin vitro. This effect was evident for cells from the primary culture. The obtained results confirm the importance of primary cells models in studies on the efficacy of experimental cancer therapies.

Human NADPH-P450 oxidoreductase modulates the level of cytochrome P450 CYP2D6 holoprotein via haem oxygenase-dependent and -independent pathways

2001 ◽  
Vol 356 (2) ◽  
pp. 613-619 ◽  
Author(s):  
Shaohong DING ◽  
Denggao YAO ◽  
Yusuf Y. DEENI ◽  
Brian BURCHELL ◽  
C. Roland WOLF ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Cell Line ◽  
Combined Treatment ◽  
Chinese Hamster ◽  
Radical Scavenger ◽  
P450 Oxidoreductase ◽  
Haem Oxygenase ◽  
Mechanistic Basis ◽  
Rate Limiting

NADPH-P450 oxidoreductase (CPR) is essential for the activity of cytochrome P450 (P450). Previous studies demonstrated that CPR regulates the levels of various P450 isoforms in vitro. We investigated the mechanistic basis for this regulation. By transfection of Chinese hamster ovary DUKXB11 cells we obtained the cell line DUKX/2D6, which expressed human CYP2D6, a P450 isoform. Subsequently, DUKX/2D6 cells were transfected with human CPR cDNA to generate the cell line DUKX/2D6/CPR-3. Expression of recombinant CPR decreased the level of spectrally detectable CYP2D6 holoprotein in DUKX/2D6/CPR-3 cells by 70%, whereas the level of immunodetectable apoprotein remained unchanged. Addition of the radical scavenger DMSO increased levels of CYP2D6 holoenzyme in DUKX/2D6/CPR-3 cells but not in DUKX/2D6 cells. A similar effect was noted when cells were grown in the presence of hemin. Importantly, combined treatment with DMSO and hemin increased levels of CYP2D6 holoenzyme in DUKX/2D6/CPR-3 but not in DUKX/2D6 cells even further than either treatment alone. None of these treatments affected the level of immunodetectable CYP2D6. This demonstrates that expression of CPR increases production of damaging radicals but also that CPR may alter haem homoeostasis. In agreement with this, the activity of haem oxygenase, a rate-limiting enzyme in haem metabolism, was compared with that in DUKX/DHFR control cells (expressing dihydrofolate reductase), and was 3-fold higher in DUKX/2D6/CPR-3 but similar in DUKX/2D6 cells. Furthermore, treatment of cells with sodium arsenite increased levels of haem oxygenase concomitant with a marked decrease of spectrally detectable CYP2D6 and a rise in levels of ferritin, which sequesters free iron released from the destruction of haem. These data demonstrate that CPR regulates P450 activity by supplying electrons and also by altering P450 levels via radical-and haem oxygenase-mediated pathways.

A Novel In Vitro Model for Irreversible Electroporation Based Cancer Therapies and Treatment Planning

2011 ◽  
Author(s):  
Christopher S. Szot ◽  
Christopher B. Arena ◽  
Paulo A. Garcia ◽  
Rafael V. Davalos ◽  
Joseph W. Freeman ◽  
...  
Keyword(s):  
Structural Changes ◽  
In Vitro Model ◽  
Irreversible Electroporation ◽  
Cancer Therapies ◽  
Electric Pulses ◽  
Therapy Option ◽  
Tissue Region ◽  
Vitro Model ◽  
Effective Cancer Therapy

Irreversible electroporation (IRE) is a minimally invasive, localized, non-thermal tissue ablation technique that has shown tremendous promise as an effective cancer therapy option. This procedure uses electrodes to apply a series of short-duration, high intensity electric pulses to a targeted tissue region. These pulses can produce irreversible structural changes in the cell membranes within the targeted region, generating a controllable range of cell death [1].

scholarly journals Interferon-γ enhances the antifibrotic effects of pirfenidone by attenuating IPF lung fibroblast activation and differentiation

2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Tuong N. Vu ◽  
Xuesong Chen ◽  
Hussein D. Foda ◽  
Gerald C. Smaldone ◽  
Nadia A. Hasaneen
Keyword(s):  
Combined Treatment ◽  
Gelatin Zymography ◽  
Interferon Γ ◽  
Lung Fibroblast ◽  
Lung Fibroblasts ◽  
Matrix Remodeling ◽  
Fibroblast Activation ◽  
Ifn Γ ◽  
Tgf Β1

Abstract Background Idiopathic pulmonary fibrosis (IPF) pathogenesis involves multiple pathways, and combined antifibrotic therapy is needed for future IPF therapy. Inhaled interferon-γ (IFN-γ) was recently shown to be safe and without systemic effects in patients with IPF. Aim To examine the in vitro effects of individual and combined treatment with IFN-γ and pirfenidone (PFD) on normal and IPF fibroblast activation and extracellular matrix remodeling after TGF-β1 and PDGF-BB stimulation. Methods IPF and normal human lung fibroblasts (NHLF) were treated with IFN-γ, PFD or a combination of both drugs in the presence of either TGF-β1 or PDGF-BB. The effects of TGF-β1 and PDGF-BB treatment on cell viability, proliferation, differentiation and migration were examined. The expression of collagen 1, matrix metalloproteinases (MMPs) and tissue inhibitors of MMP (TIMPs) was analyzed using qPCR, Western blotting and gelatin zymography. Total collagen content in conditioned media was also measured using a Sircol assay. Results Compared to that of PFD, the effect of IFN-γ in downregulating normal and IPF lung fibroblast differentiation to myofibroblasts in response to TGF-β1 was more potent. Importantly, the combination of IFN-γ and PFD had a possibly synergistic/additive effect in inhibiting the TGF-β1- and PDGF-BB-induced proliferation, migration and differentiation of normal and IPF lung fibroblasts. Furthermore, both drugs reversed TGF-β1-induced effects on MMP-1, − 2, − 3, − 7, and − 9, while only PFD promoted TIMP-1 and-2 expression and release. Conclusions Our findings demonstrate that the antifibrotic effects of IFN-γ and PFD on normal and IPF lung fibroblasts are different and complementary. Combination therapy with inhaled IFN-γ and PFD in IPF is promising and should be further explored in IPF clinical trials.

scholarly journals Effect of Benzaldehyde on Survivals of Chinese Hamster V-79 Cells in Vitro from The Combined Treatment with Hyperthermia.

1998 ◽  
Vol 14 (2) ◽  
pp. 125-129 ◽  
Author(s):  
EIICHI KANO ◽  
KAZUMI NITTA ◽  
SACHIKO HAYASHI ◽  
MASANORI HATASHITA ◽  
NAHOKO KOKUBO ◽  
...  
Keyword(s):  
Combined Treatment ◽  
Chinese Hamster

Effect of electrical stimulation on biological cells by capacitive coupling – an efficient numerical study considering model uncertainties

2020 ◽  
Author(s):  
Julius Zimmermann ◽  
RIchard Altenkirch ◽  
Ursula van Rienen
Keyword(s):  
Electrical Stimulation ◽  
Cell Membrane ◽  
Electric Fields ◽  
Biological Samples ◽  
Numerical Study ◽  
Cell Activity ◽  
Cellular Level ◽  
Capacitive Coupling ◽  
Fe Method

Electrical stimulation of biological samples such as tissues and cell cultures attracts growing attention due to its capability of enhancing cell activity, proliferation and differentiation. <br>Eventually, profound knowledge of the underlying mechanisms paves the way for innovative therapeutic devices. <br>Capacitive coupling is one option of delivering electric fields to biological samples and has advantages with regard to biocompatibility.<br>However, the mechanism of interaction is not well understood.<br>Experimental findings could be related to voltage-gated channels, which are triggered by changes of the transmembrane potential (TMP).<br>Numerical simulations by the Finite Element method (FEM) provide a possibility to estimate the TMP.<br>For realistic simulations of <i>in vitro</i> electric stimulation experiments, a bridge from the mesoscopic level down to the cellular level has to be found.<br>A special challenge poses the ratio between the cell membrane (a few <i>nm</i>) and the general setup (some <i>cm</i>).<br>Hence, a full discretization of the cell membrane becomes prohibitively expensive for 3D simulations.<br>We suggest using an approximate FE method that makes 3D multi-scale simulations possible.<br>Starting from an established 2D model, the chosen method is characterized and applied to realistic <i>in vitro</i> situations.<br>A to date not investigated parameter dependency is included and tackled by means of Uncertainty Quantification (UQ) techniques.<br>It reveals a strong, frequency-dependent influence of uncertain parameters on the modeling result.<br><br>

The Attachment of V79 and Human Periodontal Ligament Fibroblasts on Periodontally Involved Root Surfaces Following Treatment with EDTA, Citric Acid, or Tetracycline HCL: An SEM in vitro Study

2006 ◽  
Vol 7 (1) ◽  
pp. 44-59 ◽  
Author(s):  
R. Viswa Chandra ◽  
Ganesh Chandra Jagetia ◽  
K. Mahalinga Bhat
Keyword(s):  
Citric Acid ◽  
Periodontal Ligament ◽  
Chinese Hamster ◽  
Lung Fibroblasts ◽  
Control Group ◽  
Periodontal Ligament Fibroblasts ◽  
Human Periodontal Ligament Fibroblasts ◽  
Root Surfaces ◽  
Tetracycline Hcl

Abstract Objective The present in vitro study has been designed to establish and compare the effects of citric acid, EDTA, and tetracycline HCl on human periodontally diseased roots on the structure, attachment, and orientation of V79 (primary Chinese hamster lung fibroblasts) cells and human periodontal ligament fibroblasts (HPDL). Materials and Methods Commercially availableV79 cells and HPDL derived from healthy human third molars were used in this study. These fibroblasts were left in solution for seven days in order to attain confluence. Forty single-rooted teeth were obtained from patients diagnosed with periodontitis. The crown part was removed under constant irrigation and the root was split vertically into two equal halves, thus, yielding 80 specimens. Following scaling and root planing, the specimens were washed with phosphate buffered saline (PBS) and kept in 50 μg/ml gentamycin sulphate solution for 24 hours. The root pieces were then treated as follows: citric acid at pH 1, 24% EDTA, or with a 10% solution of tetracycline HCl and were then placed in V79 fibroblast cultures and HPDL cultures. The specimens were harvested after four weeks and were fixed in 2.5% glutaraldehyde in PBS before preparation for scanning electron microscopy (SEM). Results The behavior of V79 cells was similar to that of human periodontal ligament cells on root conditioned surfaces. V79 and HPDL showed a healthy morphology on root surfaces treated with citric acid and EDTA and a relatively unhealthy appearance on root surfaces treated with tetracycline HCl and distilled water (control group). Conclusion The results suggest the use of citric acid and EDTA as root conditioning agents favorably affects the migration, attachment, and morphology of fibroblasts on human root surfaces, which may play a significant role in periodontal healing and regeneration. Citation Chandra RV, Jagetia GC, Bhat KM. The Attachment of V79 and Human Periodontal Ligament Fibroblasts on Periodontally Involved Root Surfaces Following Treatment with EDTA, Citric Acid, or Tetracycline HCL: An SEM in vitro Study. J Contemp Dent Pract 2006 February;(7)1:044-059.

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