Can phenotypic data complement our understanding of antimycobacterial effects for drug combinations?

Abstract Objectives To demonstrate how phenotypic cell viability data can provide insight into antimycobacterial effects for the isoniazid/rifampicin treatment backbone. Methods Data from a Mycobacterium komossense hollow-fibre infection model comprising a growth control group, rifampicin at three different exposures (Cmax = 0.14, 0.4 and 1.47 mg/L with t½ = 1.57 h and τ = 8 h) and rifampicin plus isoniazid (Cmax rifampicin = 0.4 mg/L and Cmax isoniazid = 1.2 mg/L with t½ = 1.57 h and τ = 8 h) were used for this investigation. A non-linear mixed-effects modelling approach was used to fit conventional cfu data, quantified using solid-agar plating. Phenotypic proportions of respiring (alive), respiring but with damaged cell membrane (injured) and ‘not respiring’ (dead) cells data were quantified using flow cytometry and Sytox Green™ (Sigma–Aldrich, UK) and resazurin sodium salt staining and fitted using a multinomial logistic regression model. Results Isoniazid/rifampicin combination therapy displayed a decreasing overall antimicrobial effect with time (θTime1/2 = 438 h) on cfu data, in contrast to rifampicin monotherapy where this trend was absent. In the presence of isoniazid a phenotype associated with cell injury was displayed, whereas with rifampicin monotherapy a pattern of phenotypic cell death was observed. Bacterial killing onset time on cfu data correlated negatively (θTime50 = 28.9 h, θLAGRIF50 = 0.132 mg/L) with rifampicin concentration up to 0.165 mg/L and this coincided with a positive relationship between rifampicin concentration and the probability of phenotypic cell death. Conclusions Cell viability data provide structured information on the pharmacodynamic interaction between isoniazid and rifampicin that complements the understanding of the antibacillary effects of this mycobacterial treatment backbone.

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Lactobacillus casei CRL 431 improves endothelial and platelet functionality in a pneumococcal infection model

Beneficial Microbes ◽

10.3920/bm2018.0099 ◽

2019 ◽

Vol 10 (5) ◽

pp. 533-541

Author(s):

C. Haro ◽

M. Medina

Keyword(s):

Cell Death ◽

Lactobacillus Casei ◽

Pneumococcal Infection ◽

Probiotic Strain ◽

Endothelial Activation ◽

Control Group ◽

Infection Model ◽

Soluble Adhesion Molecules ◽

Induce Platelet Aggregation ◽

Platelet Counts

Streptococcus pneumoniae is able to activate coagulation and induce platelet aggregation, both of which are typical responses to systemic inflammation. The interactions between inflammation and coagulation and between soluble adhesion molecules and endothelial cells are important in the pathogenesis of an unbalanced haemostatic system. Therefore, an exaggerated and/or insufficiently controlled haemostatic activity may appreciably contribute to the severity of the disease. The aim of the present study was to evaluate the effect of the oral administration of Lactobacillus casei CRL 431 on platelet and endothelial activation mechanisms in a respiratory pneumococcal infection model in mice. S. pneumoniae induced an increase in platelet counts and enhanced the expression of P-selectin in control group, with higher endothelial activation in lung shown by the increase in von Willebrand factor (vWF) and vascular cell adhesion molecule 1 (VCAM-1) expression. Also, infection induced a decrease in CXCR-4 leukocytes, increased expression in annexinV and cell death at the pulmonary level and decreased antithrombin levels in bronchoalveolar lavage. In contrast, L. casei mice restored platelet counts, favoured faster P-selectin expression, lower vWF levels and VCAM-1 expression than control group. Also, L. casei induced higher levels of annexinV expression and lower cell death in the lung. Moreover, it was able to modulate antithrombin levels within the normal range, which would indicate lower coagulation activation and a protective effect locally exerted by L. casei. In this work, the ability of L. casei to favourably modulate platelet and endothelial functionality during a pulmonary infection with S. pneumoniae was demonstrated. Our findings offer a promising perspective for the use of this probiotic strain in the prevention of thrombotic complications associated with pneumococcal pneumonia, especially in at-risk patients. In addition, the use of L. casei would provide novel alternatives for the prevention and treatment of thrombosis associated with various diseases.

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Oxytocin Promotes C6 Glial Cell Death and Aggravates Hydrogen Peroxide-induced Oxidative Stress

10.36922/itps.v3i1.939 ◽

2020 ◽

pp. 27-33

Author(s):

Ahmet Sevki Taskiran ◽

Merve Ergul

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Cell Death ◽

Cell Viability ◽

Glial Cell ◽

Glial Cells ◽

Control Group ◽

C6 Cells ◽

Cox 2 ◽

Cox 1

Background. Recent studies have shown that oxytocin plays a vital role in neurons and glial cells. However, its effect on hydrogen peroxide (H2O2)-induced oxidative stress as well as cyclooxygenase-1 (COX-1) and COX-2 in glial cells are still unclear. Objective. This study aims to examine the effect of oxytocin on glial cell viability, oxidative stress, COX-1, and COX-2 in C6 glial cells after exposure to H2O2. Methods. In this study, C6 glioma cell line was used to study the effect of oxytocin on the glial cell in four cell groups. The control group was untreated. Cells in the H2O2 group were treated with 0.5 mM H2O2 for 24 h. Cells in the oxytocin group were treated with various concentrations (0.25, 0.5, 1, and 2 μg/mL) of oxytocin for 24 h. Cells in the oxytocin+H2O2 group were pre-treated with various concentrations (0.25, 0.5, 1, and 2 μg/mL) of oxytocin for 1 h before 24-h exposure to 0.5 mM H2O2. Cell viability was evaluated using XTT assay. Total antioxidant status and total oxidant status (TOS), COX-1, and COX-2 levels in the cells were measured by commercial kits. Results. Oxytocin with various concentrations significantly decreased the viability of C6 cells after H2O2-induced oxidative stress (P < 0.01). It also significantly increased the levels of TOS, COX-1, and COX-2 in C6 cells after H2O2-induced oxidative stress (P < 0.001). Conclusion. Oxytocin increases glial cell death after H2O2-induced oxidative damage in C6 cells, along with upregulation of COX-1 and COX-2 levels.

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FTY720 Induces Apoptosis and Autophagy in Multiple Myeloma Cells

Blood ◽

10.1182/blood.v118.21.5103.5103 ◽

2011 ◽

Vol 118 (21) ◽

pp. 5103-5103

Author(s):

Aijun Liao ◽

Rong Hu ◽

Qihui Zhao ◽

Huihan Wang ◽

Yingchun Li ◽

...

Keyword(s):

Multiple Myeloma ◽

Cell Death ◽

Cell Viability ◽

Cell Line ◽

Control Group ◽

Bafilomycin A1 ◽

Antiapoptotic Proteins ◽

Dependent Cell ◽

Induced Apoptosis ◽

Dose Dependent

Abstract Abstract 5103 Despite recent treatment advances, multiple myeloma(MM) remains incurable. FTY720 has initially been used as an immunosuppressant and is promising in treating relapsing/remitting multiple sclerosis. FTY720 has also been studied in several hematological malignancies including MM, but there are no reports about autophagy induced by FTY720 in MM. Therefore, we assessed the efficacy of FTY720 on MM cell line U266 and investigated the associated mechanisms of cell death. First we examined whether or not FTY720 induce cell death in U266 cells. We treated U266 cells with different dosage of FTY720 and then cell viability was measured at 24 hours by CCK-8 assay. The results showed cell viability started to reduce at 5uM and reach to 15% at 20uM, suggesting FTY720 affects cell survival in MM cell line. We then measured cell viability at 2□ A6□ A24 hours at fix concentration of 20.0 mM, 50% cells were killed even at 2 hours as compared with control group. Cell apoptosis was also measured by flow cytometry(FCM). Apoptosis rate(%) increased in a dose-dependent manner after FTY720 treatment. The percent Annexin+/7AAD- cells in the control group remained constant 5% and 15% to 30% in the drug treated time course experiments. The cell viability and apoptosis induced by FTY720 showed a dose- and time-dependant manner. Z-VAD-fmk, a pancaspase inhibitor, could rescue cell death caused by FTY720 and exposure of U266 cells to different concentration of FTY720 induced cleavage of caspase-3. We conclude FTY720 can induce a caspase-dependent cell death in U266 cells. Mcl-1, survivin, bcl-2 and Bid are antiapoptotic proteins and degradation of these proteins are required for the induction of apoptosis. In our study, the expression of Mcl-1, survivin and bcl-2 were reduced after the treatment which confirmed FTY720 induced apoptosis by down-regulating the expression of antiapoptotic proteins. Consistent with this, cleavage of Bid was also increased after the FTY720 treatment. Autophagy is another cell death pathway characterized by intracellular degradation system that delivers cytoplasmic constitutions to the lysosome. Conversion of microtubule-associated protein 1 light chain 3 (LC3)-I to LC3-II is a marker of autophagosome degradation. We observed increasing amount of conversion from LC3-I to LC3-II after FTY720 treatment, suggesting that FTY720 could induce autophagy in U266 cells. This conversion is dose-dependent. Interestingly, LC3-II accumulated in the presence of Bafilomycin A1, an inhibitor of autophagosome-lysosome fusion and LC3-II degradation, in cells treated with FTY720, confirmed that autophagic flux was induced by FTY720. To understand the role of apoptosis and autophagy played in the FTY720 induced cell death, we examined cell viability and apoptosis after FTY720 treatment in the presence or absence of Bafilomycin A1. Cell viability was higher and apoptosis was lower in the presence of Bafilomycin A1 after FTY720 treatment, indicating that Bafilomycin A1 could rescue cell death and apoptosis caused by FTY720. The results suggested that autophagy and apoptosis synergized to induce MM cell death after FTY720 treatment. We found Bafilomycin A1 rescued FTY720 induced cell death is dependent on the accumulation of anti-apoptotic protein Mcl-1 and survivin, it is possible that autophagy helps to degrade the anti-apoptotic proteins in the lysosome and synergize the cell death induced by FTY720. To investigate the mediators of FTY720-induced apoptosis and autophagy, we examined reactive oxygen species (ROS), which plays an important role in regulating both apoptosis and autophagy. Potential antioxidants N-acetyl-L-cysteine (NAC) and Tiron both could rescue apoptosis induced by FTY720, suggesting that FTY720 induced apoptosis via the activation of ROS. Furthermore, both NAC and Tiron blocked the conversion of LC3-I to LC3-II, indicating that FTY720 leads to ROS production, which results in autophagosome development and autolysosomal degradation. These studies indicate autophagy induced by FTY720 contributes to cell death in U266 cells and suggest that the function of FTY720 maybe can benefit from autophagy enhancement. To this regard, the ability of FTY720 to induce autophagy-dependent cell death in U266 cells, if confirmed in vivo, may represent a relatively selective therapy for the treatment of MM. Disclosures: No relevant conflicts of interest to declare.

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1,8-Cineol Attenuated AΒ25-35-Induced PC12 Cell Injury through Reducing Caspase 3 Expression and NO Production

Medical Research ◽

10.6913/mrhk.201912_1(1).0002 ◽

2019 ◽

Vol 1 (1) ◽

pp. 12-17

Author(s):

Ze-qin Zhang ◽

Hai-jian Li ◽

Wan-zhong Li ◽

Lin Wang ◽

Zhen-zhen Li ◽

...

Keyword(s):

Protein Expression ◽

Cell Viability ◽

Pc12 Cells ◽

Pc12 Cell ◽

Caspase 3 ◽

Cell Injury ◽

Control Group ◽

No Production ◽

Detection Assay

Objective To investigate the effect of 1,8-cineol on caspase 3 expression and NO production induced by Aβ25-35 in PC12 cells. Methods PC12 cells were cultured in vitro, and cell injury was induced by Aβ25-35 with a concentration of 20 μM. 1,8-cineol (1, 3, 10 μM) was pretreated before Aβ25-35 treatment. PC12 cell viability was evaluated by MTT detection assay. Caspase 3 protein expression was detected by Western blotting. The level of NO production in PC12 cells was measured using ELISA detection assay kit. Results In cultured PC12 cells in vitro, MTT results showed that 20 μM of Aβ25-35 reduced cell viability significantly compared with control group. The cell viability was increased by pretreatment with 1,8-cineol with concentrations of 3 and 10 μM compared with Aβ25-35 only group. Western blotting results showed compared with control group, caspase 3 expression was increased significantly in 20 μM Aβ25-35 group. Compared with Aβ25-35 group, 1,8-cineol of 3 and 10 μM group reduced caspase 3 protein expression significantly. The level of NO production in PC12 cells was increased significantly, which was decreased by pretreatment with 3 and 10 μM of 1,8-cineol. Conclusions: Our results revealed a protective effect of 1,8-cineol on Aβ25-35 induced PC12 cell injury through inhibition of caspase 3 expression and NO production.

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MicroRNA-194 Regulates Lipopolysaccharide-Induced Cell Viability by Inactivation of Nuclear Factor-κ B Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000484453 ◽

2017 ◽

Vol 43 (6) ◽

pp. 2470-2478 ◽

Cited By ~ 6

Author(s):

Fei Xie ◽

Lei Yang ◽

Lili Han ◽

Bin Yue

Keyword(s):

Cell Viability ◽

Cell Apoptosis ◽

Cell Injury ◽

Transfection Efficiency ◽

Human Fibroblasts ◽

Control Group ◽

Nuclear Factor ◽

Effective Administration ◽

Protein Expressions ◽

Underlying Mechanisms

The present study explored the functional role of microRNA (miR)-194 in lipopolysaccharide (LPS) induced lung cell injury, along with the underlying mechanisms and to reveal the potential role in infantile pneumonia. Human fibroblasts WI38 cells were transfected with miR-194 mimic or miR-194 inhibitor, and the transfection efficiency was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Thereafter, the cells were treated with or without LPS, and then cell viability, cell apoptosis and mRNA and protein expressions of key proteins of nuclear factor kappa B (NF-κB) pathway including inhibitor of NF-κB (IκB) α, p-65, and B-cell CLL/lymphoma (Bcl) 3 were analyzed. Results showed that overexpression and suppression of miR-194 was effective. Administration of LPS significantly decreased the cell viability and statistically promoted the percentages of apoptotic cells and increased the mRNA and protein expressions of p-65 and Bcl-3 but downregulated IκBα compared to the control group (P < 0.05 or P < 0.01). LPS in combination with miR-194 suppression further enhanced the effects of LPS on cell viability and cell apoptosis compared to the LPS group (P < 0.05). In contrast, LPS in combination with miR-194 overexpression observably reversed the effects of LPS on cell viability, cell apoptosis and mRNA and protein expressions of the key proteins (P < 0.05 or P < 0.01). In conclusion, miR-194 increases the LPS-induced the inhibition of cell viability and increasing of the cell apoptosis by inhibition of NF-κB pathway in WI38 cells. MiR-194 might be a potential targeted therapy for treatment of infantile pneumonia.

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Valspodar-modulated chemotherapy in human ovarian cancer cells SK-OV-3 and MDAH-2774

Bosnian Journal of Basic Medical Sciences ◽

10.17305/bjbms.2019.4073 ◽

2019 ◽

Author(s):

Maciej Zalewski ◽

Julita Kulbacka ◽

Jolanta Saczko ◽

Małgorzata Drag-Zalesinska ◽

Anna Choromanska

Keyword(s):

Oxidative Stress ◽

Ovarian Cancer ◽

Cell Death ◽

Multidrug Resistance ◽

Cell Viability ◽

Cancer Cells ◽

Gynecologic Oncology ◽

Ovarian Cancer Cells ◽

Control Group ◽

Human Ovarian Cancer

Overcoming drug resistance in ovarian cancer is the overarching goal in gynecologic oncology. One way to increase drug cytotoxicity without increasing the drug dose is to simultaneously apply multidrug resistance modulator. Valspodar is the second generation P-glycoprotein 1 modulator capable of reversing multidrug resistance in different cancers. In this study, we evaluated the effect of valspodar and cisplatin co-treatment on cell viability, cell death and oxidative status in ovarian cancer cells. The two cisplatin-resistant human ovarian cancer cell lines SK-OV-3 and MDAH-2774 were treated with cisplatin, valspodar, or cisplatin+valspodar for 24 and 48 hours. Untreated cells were used as control group. Cell viability was evaluated by MTT assay. Cell death was assessed by TUNEL and comet assay. Lipid peroxidation (malondialdehyde) and protein thiol groups were analyzed as oxidative stress markers. The expression of mitochondrial superoxide dismutase (MnSOD) was assessed by immunocytochemistry. Valspodar effectively reduced the resistance of SK-OV-3 cells to cisplatin, as demonstrated by increased oxidative stress, decreased cell viability and increased apoptosis in SK-OV-3 cells co-treated with valspodar and cisplatin compared to other groups. However, valspodar did not affect the resistance of MDAH-2774 cells to cisplatin. Stronger staining for MnSOD in MDAH-2774 vs. SK-OV-3 cells after co-treatment with cisplatin and valspodar may determine the resistance of MDAH-2774 cell line to cisplatin.

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Stew in its Own Juice: Protein Homeostasis Machinery Inhibition Reduces Cell Viability in Multiple Myeloma Cell Lines

Current Molecular Medicine ◽

10.2174/1566524019666190305134441 ◽

2019 ◽

Vol 19 (2) ◽

pp. 112-119 ◽

Cited By ~ 1

Author(s):

Mariana B. de Oliveira ◽

Luiz F.G. Sanson ◽

Angela I.P. Eugenio ◽

Rebecca S.S. Barbosa-Dantas ◽

Gisele W.B. Colleoni

Keyword(s):

Multiple Myeloma ◽

Cell Death ◽

Cell Viability ◽

Cell Lines ◽

Treatment Options ◽

Compensatory Mechanism ◽

Protein Homeostasis ◽

Protein Response ◽

Rpmi 8226

Introduction:Multiple myeloma (MM) cells accumulate in the bone marrow and produce enormous quantities of immunoglobulins, causing endoplasmatic reticulum stress and activation of protein handling machinery, such as heat shock protein response, autophagy and unfolded protein response (UPR).Methods:We evaluated cell lines viability after treatment with bortezomib (B) in combination with HSP70 (VER-15508) and autophagy (SBI-0206965) or UPR (STF- 083010) inhibitors.Results:For RPMI-8226, after 72 hours of treatment with B+VER+STF or B+VER+SBI, we observed 15% of viable cells, but treatment with B alone was better (90% of cell death). For U266, treatment with B+VER+STF or with B+VER+SBI for 72 hours resulted in 20% of cell viability and both treatments were better than treatment with B alone (40% of cell death). After both triplet combinations, RPMI-8226 and U266 presented the overexpression of XBP-1 UPR protein, suggesting that it is acting as a compensatory mechanism, in an attempt of the cell to handle the otherwise lethal large amount of immunoglobulin overload.Conclusion:Our in vitro results provide additional evidence that combinations of protein homeostasis inhibitors might be explored as treatment options for MM.

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CBIO-07. CELL DEATH INDUCED BY AN OSCILLATING MAGNETIC FIELD IN PATIENT DERIVED GLIOBLASTOMA CELLS IS MEDIATED BY REACTIVE OXYGEN SPECIES

Neuro-Oncology ◽

10.1093/neuonc/noaa215.067 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii17-ii17

Author(s):

Shashank Hambarde ◽

Martyn Sharpe ◽

David Baskin ◽

Santosh Helekar

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Cell Viability ◽

Cortical Neurons ◽

Reactive Oxygen ◽

Great Promise ◽

Antioxidant Vitamin ◽

Ros Generation ◽

Oxygen Species ◽

Novel Therapy

Abstract Noninvasive cancer therapy with minimal side effects would be ideal for improving patient outcome in the clinic. We have developed a novel therapy using strong rotating magnets mounted on a helmet. They generate oscillating magnetic fields (OMF) that penetrate through the skull and cover the entire brain. We have demonstrated that OMF can effectively kill patient derived glioblastoma (GBM) cells in cell culture without having cytotoxic effects on cortical neurons and normal human astrocytes (NHA). Exposure of GBM cells to OMF reduced the cell viability by 33% in comparison to sham-treated cells (p< 0.001), while not affecting NHA cell viability. Time lapse video-microscopy for 16 h after OMF exposure showed a marked elevation of mitochondrial reactive oxygen species (ROS), and rapid apoptosis of GBM cells due to activation of caspase 3. Addition of a potent antioxidant vitamin E analog Trolox effectively blocked OMF-induced GBM cell death. Furthermore, OMF significantly potentiated the cytotoxic effect of the pro-oxidant Benzylamine. The results of our studies demonstrate that OMF-induced cell death is mediated by ROS generation. These results demonstrate a potent oncolytic effect on GBM cells that is novel and unrelated to any previously described therapy, including a very different mechanism of action and different technology compared to Optune therapy. The effect is very powerful, and unlike Optune, can be seen within hours after initiation of treatment. We believe that this technology holds great promise for new, effective and nontoxic treatment of glioblastoma.

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MicroRNA-513b-5p targets COL1A1 and COL1A2 associated with the formation and rupture of intracranial aneurysm

Scientific Reports ◽

10.1038/s41598-021-94116-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Zheng Zheng ◽

Yan Chen ◽

Yinzhou Wang ◽

Yongkun Li ◽

Qiong Cheng

Keyword(s):

Cell Death ◽

Intracranial Aneurysm ◽

Luciferase Reporter Assay ◽

Luciferase Reporter ◽

Control Group ◽

Type I ◽

Mrna Levels ◽

Reporter Assay ◽

Over Expression ◽

Tnf Α

AbstractCollagen-type I alpha 1 chain (COL1A1) and COL1A2 are abnormally expressed in intracranial aneurysm (IA), but their mechanism of action remains unclear. This study was performed to investigate the mechanism of COL1A1 and COL1A2 affecting the occurrence and rupture of IA. Quantitative real-time polymerase chain reaction was used to measure the expression of hsa-miR-513b-5p, COL1A1, COL1A2, TNF-α, IL-6, MMP2, MMP3, MMP9 and TIMP4 in patients with ruptured IA (RA) (n = 100), patients with un-ruptured IA (UA) (n = 100), and controls (n = 100). Then, human vascular smooth muscle cells (HASMCs) were cultured, and dual luciferase reporter assay was performed to analyse the targeting relationship between miR-513b-5p and COL1A1 or COL1A2. The effects of the miR-513b-5p mimic and inhibitor on the proliferation, apoptosis, and death of HASMC and the RIP1-RIP3-MLKL and matrix metalloproteinase pathways were also explored. The effect of silencing and over-expression of COL1A1 and COL1A2 on the role of miR-513b-5p were also evaluated. Finally, the effects of TNF-α on miR-513b-5p targeting COL1A1 and COL1A2 were tested. Compared with those in the control group, the serum mRNA levels of miR-513b-5p, IL-6 and TIMP4 were significantly decreased in the RA and UA groups, but COL1A1, COL1A2, TNF-α, IL-1β, MMP2, MMP3 and MMP9 were significantly increased (p < 0.05). Compared with those in the UA group, the expression of COL1A1, COL1A2, TNF-α, IL-1β and MMP9 was significantly up-regulated in the RA group (p < 0.05). Results from the luciferase reporter assay showed that COL1A1 and COL1A were the direct targets of miR-513b-5p. Further studies demonstrated that miR-513b-5p targeted COL1A1/2 to regulate the RIP1-RIP3-MLKL and MMP pathways, thereby enhancing cell death and apoptosis. Over-expression of COL1A1 or COL1A2, rather than silencing COL1A1/2, could improve the inhibitory effect of miR-513b-5p on cell activity by regulating the RIP1-RIP3-MLKL and MMP pathways. Furthermore, over-expression of miR-513b-5p and/or silencing COL1A1/2 inhibited the TNF-α-induced cell proliferation and enhanced the TNF-α-induced cell death and apoptosis. The mechanism may be related to the inhibition of collagen I and TIMP4 expression and promotion of the expression of RIP1, p-RIP1, p-RIP3, p-MLKL, MMP2 and MMP9. MiR-513b-5p targeted the inhibition of COL1A1/2 expression and affected HASMC viability and extracellular mechanism remodelling by regulating the RIP1-RIP3-MLKL and MMP pathways. This process might be involved in the formation and rupture of IA.

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Cytotoxicity assessment of different doses of ozonated water on dental pulp cells

BMC Oral Health ◽

10.1186/s12903-021-01392-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Ferdiye Küçük ◽

Sibel Yıldırım ◽

Serap Çetiner

Keyword(s):

Cell Viability ◽

Repeated Measures ◽

Dental Pulp ◽

Control Group ◽

Dental Pulp Cells ◽

Time Points ◽

Significant Difference ◽

Pulp Cells ◽

Time Point ◽

Ozonated Water

Abstract Background The purpose of this study was to assess the cytotoxicity of various concentrations of ozonated water (OW) on human primary dental pulp cells. Methods Human primary dental pulp cells were isolated from exfoliated primary canine teeth of an 11-year-old patient with good systemic and oral health. Afterwards, cells were divided into 6 experimental groups; four groups of OW in concentrations of 2 mg/L, 4 mg/L, 8 mg/L, and 16 mg/L, untreated control group, and cell culture without cells. Cytotoxicity was evaluated after exposure for 5-min exposure using Mosmann’s Tetrazolium Toxicity (MTT) assay at 0 h and 48 h time points. Data were analyzed using a repeated measures analysis of variance and Post-hoc tests were performed using Bonferroni correction for multiple comparisons. Results All experimental groups showed proliferation at 0 h time point. However, all groups also experienced a decrease in overtime at 48 h time point (p < 0.05). At both time points 2 mg/L OW showed the highest cell viability as well as proliferation. At 0 h time point, the increase in cell viability for all experimental groups was found statistically significant when compared to positive control group (p < 0.05). At 48 h time point, although 8 mg/L and 16 mg/L OW showed statistically significant reduction in compare to 0 h time point, 2 mg/L and 4 mg/L OW groups didn’t experience any statistically significant difference (p < 0.05). Conclusion Considering our findings, due to ozonated water's induced a higher proliferation rate of dental pulp cells, indicating their biocompatibility and a possible adjuvant on irrigating agent in regenerative endodontic procedures.

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