scholarly journals Cold Atmospheric Plasma Promotes Regeneration-Associated Cell Functions of Murine Cementoblasts In Vitro

2021 ◽  
Vol 22 (10) ◽  
pp. 5280
Author(s):  
Benedikt Eggers ◽  
Jana Marciniak ◽  
James Deschner ◽  
Matthias Bernhard Stope ◽  
Alexander Mustea ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Viability ◽  
Atmospheric Plasma ◽  
Type I ◽  
Mrna Levels ◽  
Xtt Assay ◽  
Ki 67 ◽  
Alizarin Red ◽  
Cold Atmospheric Plasma ◽  
Cell Functions

The aim of the study was to examine the efficacy of cold atmospheric plasma (CAP) on the mineralization and cell proliferation of murine dental cementoblasts. Cells were treated with CAP and enamel matrix derivates (EMD). Gene expression of alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (BGLAP), periostin (POSTN), osteopontin (OPN), osterix (OSX), collagen type I alpha 1 chain (COL1A1), dentin matrix acidic phosphoprotein (DMP)1, RUNX family transcription factor (RUNX)2, and marker of proliferation Ki-67 (KI67) was quantified by real-time PCR. Protein expression was analyzed by immunocytochemistry and ELISA. ALP activity was determined by ALP assay. Von Kossa and alizarin red staining were used to display mineralization. Cell viability was analyzed by XTT assay, and morphological characterization was performed by DAPI/phalloidin staining. Cell migration was quantified with an established scratch assay. CAP and EMD upregulated both mRNA and protein synthesis of ALP, POSTN, and OPN. Additionally, DMP1 and COL1A1 were upregulated at both gene and protein levels. In addition to upregulated RUNX2 mRNA levels, treated cells mineralized more intensively. Moreover, CAP treatment resulted in an upregulation of KI67, higher cell viability, and improved cell migration. Our study shows that CAP appears to have stimulatory effects on regeneration-associated cell functions in cementoblasts.

scholarly journals The synergistic effect of Canady Helios cold atmospheric plasma and a FOLFIRINOX regimen for the treatment of cholangiocarcinoma in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Olivia Jones ◽  
Xiaoqian Cheng ◽  
Saravana R. K. Murthy ◽  
Lawan Ly ◽  
Taisen Zhuang ◽  
...  
Keyword(s):  
Cell Viability ◽  
Chemotherapy Regimen ◽  
Synergetic Effect ◽  
Atmospheric Plasma ◽  
High Recurrence Rate ◽  
Cold Atmospheric Plasma ◽  
Tract Cancer ◽  
Anticancer Properties ◽  
Potential Interactions

AbstractCholangiocarcinoma (CCA) is a rare biliary tract cancer with a low five-year survival rate and high recurrence rate after surgical resection. Currently treatment approaches include systemic chemotherapeutics such as FOLFIRINOX, a chemotherapy regimen is a possible treatment for severe CCA cases. A limitation of this chemotherapy regimen is its toxicity to patients and adverse events. There exists a need for therapies to alleviate the toxicity of a FOLFIRINOX regimen while enhancing or not altering its anticancer properties. Cold atmospheric plasma (CAP) is a technology with a promising future as a selective cancer treatment. It is critical to know the potential interactions between CAP and adjuvant chemotherapeutics. In this study the aim is to characterize the efficacy of FOLFIRINOX and CAP in combination to understand potential synergetic effect on CCA cells. FOLFIRINOX treatment alone at the highest dose tested (53.8 µM fluorouracil, 13.7 µM Leucovorin, 5.1 µM Irinotecan, and 3.7 µM Oxaliplatin) reduced CCA cell viability to below 20% while CAP treatment alone for 7 min reduced viability to 3% (p < 0.05). An analysis of cell viability, proliferation, and cell cycle demonstrated that CAP in combination with FOLFIRINOX is more effective than either treatment alone at a lower FOLFIRINOX dose of 6.7 µM fluorouracil, 1.7 µM leucovorin, 0.6 µM irinotecan, and 0.5 µM oxaliplatin and a shorter CAP treatment of 1, 3, or 5 min. In conclusion, CAP has the potential to reduce the toxicity burden of FOLFIRINOX and warrants further investigation as an adjuvant therapy.

scholarly journals Ticagrelor and clopidogrel suppress NF-κB to treat acute coronary syndrome

2019 ◽  
Author(s):  
Zhuyin Jia ◽  
Yiwei Huang ◽  
Xiaojun Ji ◽  
Jiaju Sun ◽  
Guosheng Fu
Keyword(s):  
Cell Cycle ◽  
Acute Coronary Syndrome ◽  
Cell Migration ◽  
Cell Viability ◽  
Umbilical Vein ◽  
Quantitative Polymerase Chain Reaction ◽  
Statistical Significance ◽  
Inflammatory Factors ◽  
Mrna Levels ◽  
Coronary Syndrome

Abstract Background: Inflammatory cytokines are involved in acute coronary syndrome (ACS),and NF-kB is the central regulator of inflammation. Moreover, ticagrelor and clopidogrelcan prevent thrombotic events and improve the care of patients with ACS. Thus, we speculated that ticagrelor and clopidogrel relieve ACS by regulating NF-kB pathway. Methods: After human umbilical vein endothelial cells (HUVECs) were cultured with ticagrelor or clopidogrel and given lipopolysaccharide (LPS) and CD14, the mRNA levels of related inflammatory factors, the protein level changes of molecules in the NF-kB pathway, and the changes in cell viability, apoptosis and the cell cycle, cell migration, vascular formation and other vital activities were detected using quantitative Polymerase chain reaction (qPCR), Western blotting and immunofluorescence assay, CCK8， flow cytometry, transwell assay, matrigel, respectively. All data was expressed as the mean ± S.D. The statistical significance of data was assessedby an unpaired two-tailed t-test. Results: Ticagrelor and clopidogrel can suppress the NF-kB pathway by inhibiting the phosphorylation and entry into the nucleus of p65, restraining the degradation of IKBa, improving cell viability, restoring the cell cycle, cell migration and angiogenic ability, and inhibiting apoptosis. Conclusions: Ticagrelor and clopidogrel alleviate cellular dysfunction through suppressing NF-kB signaling to treat acute coronary syndrome.

scholarly journals Involvement of BMP-15 in Glucocorticoid Actions on Ovarian Steroidogenesis by Rat Granulosa Cells

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A767-A768
Author(s):  
Chiaki Kashino ◽  
Toru Hasegawa ◽  
Yasuhiro Nakano ◽  
Nahoko Iwata ◽  
Koichiro Yamamoto ◽  
...  
Keyword(s):  
Bone Morphogenetic Proteins ◽  
Granulosa Cells ◽  
Type I ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Ovarian Steroidogenesis ◽  
Cell Functions ◽  
Camp Synthesis ◽  
Bmp Receptors

Abstract Glucocorticoid receptor (GR) are known to be expressed in the ovary and glucocorticoids are shown to exert direct effects on granulosa cell functions. In the clinical setting, menstrual abnormality, amenorrhea and hypermenorrhea can be shown in patients with glucocorticoid excess. On the other hand, glucocorticoids can also be used for the treatment of PCOS with hyperandrogenism. However, the effects of glucocorticoids on the reproductive system have not been fully elucidated. In the present study, we investigated the influence of glucocorticoids on follicular steroidogenesis using primary culture of rat granulosa cells, by focusing on the ovarian bone morphogenetic proteins (BMPs) acting as a luteinizing inhibitor. Granulosa cells isolated from female immature rats were treated with follicle-stimulating hormone (FSH) in the presence of dexamethasone (Dex) in serum-free conditions. After treatment with Dex for 48 h, the changes of estradiol (E2) and progesterone (P4) production and cAMP synthesis induced by FSH treatments were measured by ELISA. Total RNAs of granulosa cells treated with FSH, Dex and BMPs were extracted and mRNA levels of steroidogenetic factors and enzymes, BMP receptors and Id-1 were quantified by real-time RT-PCR. Phosphorylation of Smad1/5/9 induced by BMPs was evaluated by Western blotting using cell lysates in the presence or absence of Dex. As a result, it was revealed that Dex treatment decreased FSH-induced E2 production by granulosa cells. In accordance with the steroid results, Dex suppressed FSH-induced P450arom mRNA expression as well as FSH-induced cAMP synthesis by granulosa cells. By contrast, Dex treatment augmented FSH-induced P4 production by granulosa cells in a concentration-dependent manner. Dex treatment was found to enhance basal and FSH-induced mRNA levels of P4-synthetic enzymes including P450scc and 3βHSD. Of note, Dex treatment activated the BMP target gene Id-1 transcription and Smad1/5/9 phosphorylation, in particular, induced by BMP-15 among various BMP ligands including BMP-2, -4, -6, -7, -9 and -15. It was also revealed that Dex treatment increased mRNA levels of ALK-6, a type-I receptor for BMP-15, and that BMP-15 treatment in turn upregulated GR mRNA levels expressed by granulosa cells. Given that BMP-15 acts as an inhibitor for P4 production by suppressing FSH-receptor actions, it was suggested that glucocorticoid is functionally linked to the enhancement of endogenous BMP-15, leading to the negative feedback toward the P4 overproduction induced by FSH and Dex in granulosa cells. Collectively, it was revealed that glucocorticoids elicit differential effects on the ovarian steroidogenesis of E2 and P4, in which GR and BMP-15 actions are mutually enhanced in granulosa cells.

scholarly journals Sensitivity of tumor versus normal cell migration and morphology to cold atmospheric plasma‐treated media in varying culture conditions

2019 ◽  
Vol 17 (2) ◽  
pp. 1900103 ◽  
Author(s):  
Marina A. Pranda ◽  
Brittney J. Murugesan ◽  
Andrew J. Knoll ◽  
Gottlieb S. Oehrlein ◽  
Kimberly M. Stroka
Keyword(s):  
Cell Migration ◽  
Normal Cell ◽  
Culture Conditions ◽  
Atmospheric Plasma ◽  
Cold Atmospheric Plasma

scholarly journals EXTH-10. THE ACTIVATION AND SENSITIZATION OF GLIOBLASTOMA CELLS VIA COLD ATMOSPHERIC PLASMA TREATMENT

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi84-vi84
Author(s):  
Jonathan Sherman ◽  
Dayun Yan ◽  
Eda Gjika ◽  
Michael Keidar
Keyword(s):  
Cell Viability ◽  
Plasma Treatment ◽  
Cell Activation ◽  
Radiation Treatment ◽  
In Vitro Study ◽  
Atmospheric Plasma ◽  
Discharge Process ◽  
H2o2 Treatment ◽  
Cold Atmospheric Plasma

Abstract BACKGROUND Treatment of glioblastoma multiforme (GBM) continues to remain a challenge using conventional treatment. Through an in vitro study we assessed the efficacy of our novel cold atmospheric plasma technology (CAP) to sensitize GBM cells to temozolomide (TMZ). METHODS The CAP jet is formed through the discharge (Pk-Pk: 5.8 kV) between a ring grounded cathode and a central anode and with He flow through a glass tube. The discharge process is driven by an AC high voltage (3.16 kV) with a frequency of 12.5 kHz. Human glioblastoma (U87MG) cells were cultured in DMEM supplemented by 1% (v/v) penicillin and streptomycin solution and 10% (v/v) FBS. CAP was delivered to U87 cells in a 96-well plate for 1 min in combination with 10 and 15 μM H2O2. The cell viability was measured by using the MTT assay. We then tested TMZ concentrations of 10 and 50 uM. Cell viability was monitored with the Cell Titer Glo 2.0. luminescent assay. All experiments were performed in triplicate and were independently repeated at least 3 times. RESULTS We identified an activation state of U87MG cells after the plasma treatment. This activation state resulted in GBM cells sensitized to reactive species identified by decreased cell viability after treatment with H2O2 as compared to the H2O2 treatment alone (p< 0.005). In addition, the plasma-activated cells were sensitized to TMZ. Cells treated with CAP in combination with TMZ displayed decreased cell viability at TMZ concentrations of (10 uM) (p< 0.05) and (50 uM) (p< 0.005) as compared to TMZ alone. CONCLUSIONS This study demonstrates the activation phenomenon on GBM cells via direct CAP treatment. Due to this activation, the GBM cells were sensitized to both H2O2 and TMZ identified via decreased cell viability. Future work looks to assess this effect of cell activation/sensitization with chemotherapy plus radiation treatment.

scholarly journals Combination therapy of cold atmospheric plasma (CAP) with temozolomide in the treatment of U87MG glioblastoma cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Eda Gjika ◽  
Sonali Pal-Ghosh ◽  
Megan E. Kirschner ◽  
Li Lin ◽  
Jonathan H. Sherman ◽  
...  
Keyword(s):  
Combination Therapy ◽  
Cell Viability ◽  
Cellular Response ◽  
Atmospheric Plasma ◽  
Treatment Modalities ◽  
Glioblastoma Cells ◽  
M Cell ◽  
Cell Viability Assay ◽  
Cold Atmospheric Plasma

Abstract Cold atmospheric plasma (CAP) technology, a relatively novel technique mainly investigated as a stand-alone cancer treatment method in vivo and in vitro, is being proposed for application in conjunction with chemotherapy. In this study, we explore whether CAP, an ionized gas produced in laboratory settings and that operates at near room temperature, can enhance Temozolomide (TMZ) cytotoxicity on a glioblastoma cell line (U87MG). Temozolomide is the first line of treatment for glioblastoma, one of the most aggressive brain tumors that remains incurable despite advancements with treatment modalities. The cellular response to a single CAP treatment followed by three treatments with TMZ was monitored with a cell viability assay. According to the cell viability results, CAP treatment successfully augmented the effect of a cytotoxic TMZ dose (50 μM) and further restored the effect of a non-cytotoxic TMZ dose (10 μM). Application of CAP in conjunction TMZ increased DNA damage measured by the phosphorylation of H2AX and induced G2/M cell cycle arrest. These findings were supported by additional data indicating reduced cell migration and increased αvβ3 and αvβ5 cell surface integrin expression as a result of combined CAP–TMZ treatment. The data presented in this study serve as evidence that CAP technology can be a suitable candidate for combination therapy with existing chemotherapeutic drugs. CAP can also be investigated in future studies for sensitizing glioblastoma cells to TMZ and other drugs available in the market.

scholarly journals Ticagrelor and clopidogrel suppress NF-κB signaling pathway to alleviate LPS-induced dysfunction in vein endothelial cells

2019 ◽  
Author(s):  
Zhuyin Jia ◽  
Yiwei Huang ◽  
Xiaojun Ji ◽  
Jiaju Sun ◽  
Guosheng Fu
Keyword(s):  
Cell Cycle ◽  
Endothelial Cells ◽  
Cell Migration ◽  
Cell Viability ◽  
Signaling Pathway ◽  
Quantitative Polymerase Chain Reaction ◽  
Statistical Significance ◽  
Chronic Obstructive ◽  
Mrna Levels ◽  
Coronary Syndrome

Abstract Background: Ticagrelor and clopidogrel, P2Y12 receptor antagonists, can prevent thrombotic events and are used to treat cardiovascular diseases such as acute coronary syndrome and chronic obstructive pulmonary disease, in which inflammation is involved. Moreover, NF-kB is the central regulator of inflammation. Thus, we suspected that ticagrelor and clopidogrel are involved in the regulation of the NF-kB signaling pathway.Methods: After human umbilical vein endothelial cells (HUVECs) were cultured with ticagrelor or clopidogrel and given lipopolysaccharide (LPS) and CD14, the mRNA levels of related inflammatory factors, the protein level and subcellular localization of molecules in the NF-kB signaling pathway, cell viability, apoptosis and the cell cycle, cell migration, and vascular formation were detected using quantitative polymerase chain reaction (qPCR), western blotting and immunofluorescence assay, CCK8, flow cytometry, transwell assay, and matrigel, respectively. All data was expressed as the mean ± S.D. The statistical significance of data was assessed by an unpaired two-tailed t-test.Results: Ticagrelor and clopidogrel can inhibit the degradation of IKBa and phosphorylation of p65, prevent p65 from entering the nucleus, reduce the production of TNFa, IL-1, IL-8, IL-6 and IL-2, and alleviate the decrease in cell viability, cell migration and angiogenesis, the changes of cell cycle and apoptosis induced by LPS. Conclusions: Ticagrelor and clopidogrel alleviate cellular dysfunction through suppressing NF-kB signaling pathway.

scholarly journals Moringa Oleifera Leaf Extracts Protect BMSC Osteogenic Induction Following Peroxidative Damage by Activating The PI3K/Akt /Foxo1 Pathway

2020 ◽  
Author(s):  
Meiling Liu ◽  
Haifeng Ding ◽  
Hongzhi Wang ◽  
Manfeng Wang ◽  
Xiaowei Wu ◽  
...  
Keyword(s):  
Cell Viability ◽  
Osteogenic Differentiation ◽  
Moringa Oleifera ◽  
Caspase 3 ◽  
Therapeutic Effects ◽  
Mrna Levels ◽  
Leaf Extracts ◽  
Alizarin Red ◽  
Peroxidative Damage ◽  
Osteogenic Induction

Abstract Objective: We aimed to investigate the therapeutic effects of Moringa oleifera leaf extracts on osteogenic induction of rat bone marrow mesenchymal stem cells (BMSCs) following peroxidative damage and to explore the underlying mechanisms. Methods: Conditioned medium was used to induce osteogenic differentiation of BMSCs, which were treated with H2O2, Moringa oleifera leaf extracts-containing serum, or the phosphatidyl inositol-3 kinase (PI3K) inhibitor Wortmannin, alone or in combination. Cell viability was measured using the MTT assay. Cell cycle was assayed using flow cytometry. Expression levels of Akt, phosphorylated (p)Akt, Foxo1, and cleaved caspase-3 were analyzed using Western blot analysis. The mRNA levels of osteogenesis-associated genes, including alkaline phosphatase (ALP), collagen І, osteopontin (OPN), and Runx2, were detected using qRT-PCR. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels, as well as superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and ALP activity were detected using commercially available kits. Osteogenic differentiation capability was determined using alizarin red staining. Results: During osteogenic induction of rat BMSCs, H2O2 reduced cell viability and proliferation, inhibited osteogenesis, increased ROS and MDA levels, and decreased SOD and GSH-PX activity. H2O2 significantly reduced pAkt and Foxo1 expression, and increased cleaved caspase-3 levels in BMSCs. Additional treatments with Moringa oleifera leaf extracts partially reversed the H2O2-induced changes. Wortmannin partially attenuated the effects of Moringa oleifera leaf extracts on protein expression of Foxo1, pAkt, and cleaved caspase-3, as well as mRNA levels of osteogenesis-associated genes.Conclusion: Moringa oleifera leaf extracts ameliorate peroxidative damage and enhance osteogenic induction of rat BMSCs by activating the PI3K/Akt/Foxo1 pathway.

scholarly journals Cold Atmospheric Plasma Jet as a Possible Adjuvant Therapy for Periodontal Disease

Molecules ◽  
2021 ◽  
Vol 26 (18) ◽  
pp. 5590
Author(s):  
Gabriela de Morais Gouvêa Lima ◽  
Aline Chiodi Borges ◽  
Thalita Mayumi Castaldelli Nishime ◽  
Gabriela de Fatima Santana-Melo ◽  
Konstantin Georgiev Kostov ◽  
...  
Keyword(s):  
Adjuvant Therapy ◽  
Treated Group ◽  
Antimicrobial Activities ◽  
Atmospheric Plasma ◽  
Blood Agar ◽  
Xtt Assay ◽  
Significance Level ◽  
Cold Atmospheric Plasma ◽  
Tissue Recovery

Due to the limitations of traditional periodontal therapies, and reported cold atmospheric plasma anti-inflammatory/antimicrobial activities, plasma could be an adjuvant therapy to periodontitis. Porphyromonas gingivalis was grown in blood agar. Standardized suspensions were plated on blood agar and plasma-treated for planktonic growth. For biofilm, dual-species Streptococcus gordonii + P. gingivalis biofilm grew for 48 h and then was plasma-treated. XTT assay and CFU counting were performed. Cytotoxicity was accessed immediately or after 24 h. Plasma was applied for 1, 3, 5 or 7 min. In vivo: Thirty C57BI/6 mice were subject to experimental periodontitis for 11 days. Immediately after ligature removal, animals were plasma-treated for 5 min once—Group P1 (n = 10); twice (Day 11 and 13)—Group P2 (n = 10); or not treated—Group S (n = 10). Mice were euthanized on day 15. Histological and microtomography analyses were performed. Significance level was 5%. Halo diameter increased proportionally to time of exposure contrary to CFU/mL counting. Mean/SD of fibroblasts viability did not vary among the groups. Plasma was able to inhibit P. gingivalis in planktonic culture and biofilm in a cell-safe manner. Moreover, plasma treatment in vivo, for 5 min, tends to improve periodontal tissue recovery, proportionally to the number of plasma applications.

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