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OBJECTIVE: Strategies for cancer therapy involve radiation therapy (RT), which accounts for about 40% of all cancer treatment types. As to current chemotherapeutics, cancer cells also develop resistance that remains a clinical problem, such as disease recurrence. Recent studies focused on understanding the molecular mechanisms of radiation-induced cell death. Conventional RT aims at treatment with a single fraction per day of 8-30 Gy per fraction. Radiotherapy increases intracellular ceramide levels that trigger cell death. Additionally, increasing intracellular ceramide by radiation may restore therapeutic sensitivity to cancer treatments. Drugs that inhibit ceramide-metabolizing enzymes like ceramidases are expected to be radiotherapy sensitizers. MATERIALS AND METHODS: In this research, we investigated the proapoptotic effects of SRS alone and in combination with ceranib-2, a ceramidase inhibitor in human breast adenocarcinoma cells. The molecular mechanism of action of RT and ceranib-2 was investigated on MCF-7 cells exposed to 13 µM ceranib-2 for 24 hours following 20 Gy radiation using MTT, radiotherapy, and annexin-V analyses. RESULTS: Results indicated that the dose of 20 Gy radiation induces apoptosis on human breast cancer cells with and without co-treatment with ceranib-2 by causing cytotoxicity in the cells. Based on the results of ceranib-2 exposure, it can be concluded that the mechanism of action may rely on an increase of intracellular ceramides, also called apoptotic lipids. CONCLUSIONS: The study results suggest that co-treatment of human breast adenocarcinoma cells with a ceramidase inhibitor, ceranib- 2, and a high dose of radiation of 20 Gy exerted cytotoxicity and apoptosis and might be a solid, potent alternative to current therapy strategies. [ABSTRACT FROM AUTHOR]

OBJECTIVE: In the present study, we investigated the effects of 2-aminobenzothiazole application on human laryngeal carcinoma cells. MATERIALS AND METHODS: Human larynx epidermoid carcinoma (HEp-2) (ATCC® CCL-23™) cells were purchased from American Type Culture Collection (ATCC, USA). Human larynx epidermoid carcinoma HEp-2 cells were cultured in complete Dulbecco's Modified Eagle's Medium (DMEM) supplemented with fetal bovine serum (FBS) (10%) and penicillin/streptomycin (1%) in a CO2 (5%) incubator under standard cell culture conditions. 2-aminobenzothiazole was prepared, and further dilutions ranging from 3.13 to 100 μM were prepared in fresh culture DMEM. HEp-2 cells on 96 well plates were incubated with the prepared dilutions of 2-aminobenzothiazole for 24, 48, and 72 hours. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test performed cytotoxicity evaluation and viability percentages. The annexin-V staining technique detected 2-aminobenzothiazole-triggered apoptosis of HEp-2 cells. The activated caspases 3/7 on HEp-2 cells after 2-aminobenzothiazole exposure were evaluated with flow cytometric analysis. The membrane potential changing of HEp-2 cells was measured following the Muse™ MitoPotential kit manufacturer instructions. RESULTS: MTT cytotoxicity test results showed that the viability of human laryngeal carcinoma cells decreased with an increase in the application of 2-aminobenzothiazole for 24 hours. The highest growth inhibition by 2-aminobenzothiazole for short-term application of 24 hours was detected at the highest concentration of 2-aminobenzothiazole (100 µ M). The results underline that the cytotoxic effect of 2-aminobenzothiazole is dose-dependent. Cytotoxicity test results for an application time of 48 hours showed that the cytotoxicity of 2-aminobenzothiazole is dose-dependent on HEp-2 cells. The required dose of 2-aminobenzothiazole to decrease the cell viability to 50 percent has been 9-fold augmented. Annexin-V findings showed that after exposure to IC50 concentration of 2-aminobenzothiazole for 24 hours, HEp-2 cells underwent the early apoptotic stage (25.99%) and late apoptotic (16.69%), whereas 56.93% of the treated cells were alive. Only 0.39% of 2-aminobenzothiazole treated cells were necrotic. All study results showed that 2-aminobenzothiazole triggered apoptosis on HEp-2 cells with a percentage of total apoptotic cells 42.62 compared to untreated HEp-2 cells. Caspase 3/7 activation results showed that only 0.65% of control HEp-2 cells were with activated caspase 3/7, and 99.35% live cells. The analysis data from the Muse cell analyzer revealed that the percentage of cells with intact mitochondrial membranes was 21.30 after 2-aminobenzothiazole application, and 79.9% were cells with depolarized mitochondrial membranes. It has been understood that the depolarization of the inner mitochondrial membrane has been considered a dysfunction in mitochondria as a sign of apoptosis and drug toxicity. CONCLUSIONS: Based on all study findings, 2-aminobenzothiazole has cytotoxicity on human laryngeal carcinoma cells in a dose and time-dependent manner. That means that it decreased viability via inducing caspase-dependent apoptosis. Consequently, it was concluded that 2-aminobenzothiazole has good potential to lead to cytotoxicity and apoptosis on human laryngeal carcinoma cells and, after deeper in vitro and in vivo investigations, can be a good candidate for designing anticancer drugs with high efficiency. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Cancer-preventative medicines like curcumin, resveratrol, and nonsteroidal anti-inflammatory medications all have their effects modulated by ceramide. According to research, these medications raise ceramide levels in cancer cells, leading to programmed cell death. Recently, cancer research has been involved in sphingolipid metabolism. The critical molecule here is ceramide. We aimed to investigate if the inhibition of ceramidases induces death in the human renal cell carcinoma cell line. MATERIALS AND METHODS: Human kidney carcinoma A-498 (ATCC® HTB-44™) cells were used as test cells. Ceranib-2, fetal bovine serum (FBS), penicillin/streptomycin, dimethyl sulfoxide, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-2H-tetrazolium bromide and Dulbecco's Modified Eagle Medium High Glucose, caspase 3/7, annexin-V, Bcl-2 activation dual detection, and MitoPotential kits were used. 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-2H-tetrazolium bro-mide (MTT) colorimetric assay, annexin-V analy-sis, caspase 3/7 analysis, Bcl-2 activation analysis, and measurement of mitochondrial membrane potential were performed. RESULTS: MTT colorimetric assay results for 24 hours indicated that the viability of human renal cell carcinoma cells decreased compared to the control group with an increase in the applied concentration of the ceramidase inhibitor-ceranib-2. The growth inhibition by ceranib-2 for 24 hours did not decrease the viability under 50%; thus, it could not be possible to calculate the IC50 value for the short-term application of ceranib-2 for 24 hours to A-498 cells. A statistically significant decrease in cell viability was recorded at doses of 100, 50, 25, and 12.2 pM of ceranib-2, and no significant decrease was detected at the lower doses of ceranib-2. The highest inhibition caused by ceranib-2 on human renal cell carcinoma cells A-498 was detected at an application time of 72 hours. This inhibition was statistically significant for all applied doses of ceranib-2 on A-498 cells compared to untreated cells. Annexin-V technique that detects the translocation of phosphatidylserine to the outer membrane of apoptotic cells indicated that after the application of ceranib-2, apoptosis was triggered on A-498 cells with a total apoptotic profile of 12.12% compared to the untreated cells that were used as controls. Compared to untreated A-498 cells, a rise in percentage to 16.25% of cells with activated caspases 3/7 was recorded after applying IC50 concentration of ceranib-2 on A-498 cells for 48 hours. CONCLUSIONS: The results of our study indicated that the application of ceramidase inhibitor, ceranib-2 on human renal cell carcinoma A-498 cells cause cytotoxicity, antiproliferative, growth inhibitory, and apoptotic efficacies in a dose and time-dependent manner probably via inhibiting the acid ceramidases that hydrolyze ceramides that induce cell death. For further conclusions, more mechanical, pharmacokinetic, and pharmaceutic, as well as in vitro and in vivo anti-cancer activity investigations are required. [ABSTRACT FROM AUTHOR]

OBJECTIVE: In the present study, we investigated the effects of Ceramide C2 application on human laryngeal carcinoma cells. MATERIALS AND METHODS: Human larynx epidermoid carcinoma HEp-2 (ATCC® CCL-23™) cells were purchased from the American Type Culture Collection (ATCC, USA). Human larynx epidermoid carcinoma HEp-2 cells were cultured in complete Dulbecco's Modified Eagle's Medium (DMEM) supplemented with fetal bovine serum (FBS) (10%) and penicillin/streptomycin (1%) in a CO2 (5%) incubator under standard cell culture conditions. Ceramide C2 was prepared, and further dilutions ranging from 3.13 to 100 MM were prepared in a fresh culture medium. Cells on 96 well plates were exposed to the prepared concentrations of ceramide C2 for 24 and 48 hours. Cytotoxicity evaluation was performed by MTT. Apoptosis profiles of HEp-2 cells were detected by annexin-V analysis. The activated caspases 3/7 on HEp-2 cells after ceramide C2 exposure were evaluated with flow cytometric analysis. The morphological changes on HEp-2 cells caused by ceramide C2 were evaluated by staining with phalloidine and acridine orange via confocal microscopy. For the Wound Healing Assay, HEp-2 cells were cultured in 6 well-plates until they became confluent. RESULTS: MTT cytotoxicity test findings revealed that the viability of human laryngeal carcinoma cells decreased with the increased application of ceramide C2 for 24 hours compared to untreated (control) cells. The highest growth inhibition by ceramide C2 for short-term application for 24 hours was detected at the highest concentration of ceramide C2 (100 yM). Annexin-V findings showed that 98.97 of HEp-2 cells were alive, and 1.63% were detected as early apoptosis for the control group. The results showed that ceramide C2 triggered apoptosis on HEp-2 cells with a percentage of total apoptotic cells of 61,40 com pared to untreated HEp-2 cells. Cysteine proteases (caspases) 3/7 activation percentages of HEp-2 cells exposed to ceramide C2 for 24 hours were compared to control cells, and the morphology of HEp-2 cells was changed with clear apoptotic signs that underlined the cytotoxicity and pro-apoptotic activity of ceramide C2. Scratch Assay assessed the migration capability of HEp-2 cells before and after the exposure to ceramide C2. It showed that ceramide C2 reduced human laryngeal carcinoma cells' migration capability and proliferation for 24 hours. CONCLUSIONS: Based on all study findings, it can be considered that short-chain ceramide C2 exerted cytotoxicity on human laryngeal carcinoma cells in a dose and time-dependent manner and reduced the viability via inducing caspase-dependent apoptosis. The overall effect might be derived from the elevated intracellular ceramide levels by the exogenous application of ceramide C2. Consequently, it was concluded that ceramide C2 has good potential to cause cytotoxicity and apoptosis in human laryngeal carcinoma cells and, after deeper in vitro and in vivo investigations, can be a good candidate for designing anti-cancer drugs with high efficiency. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Dexpanthenol is an ingredient in multiple topical pharmaceutical preparations thanks to its high penetration and localized concentration. It is included in many ointments or lotions for dermatological use, assisting in healing and reducing pruritus. Vaseline is a synthetic product obtained by distilling crude oil. It is commercially available in several grades. The study presented here examined how topically applied agents (dexpanthenol or vaseline) affect nasal epithelial cells in culture. In particular, the study aimed to identify any alterations to epithelial cells which might indicate toxicity. MATERIALS AND METHODS: The nasal epithelial cells used were sourced from mucosal tissue fragments left over the following septorhinoplasty on five patients not suffering from rhinosinusitis. The first step was to dissect the mucosal fragments into smaller pieces on a sterilized Petri dish. These fragments were then placed into the DMEM-F12 cell culture medium, which had been freshly prepared. The dexpanthenol and vaseline were diluted in dimethylsulfoxide (DMSO) to a concentration of 5 mg/ mL. The cells in the wells were exposed to varying concentrations of dexpanthenol or vaseline. The actual concentration of the test reagent to which the epithelial cells were exposed ranged from 0.15 mg/mL to 5 mg/mL. The exposure period was 24 hours. The cells were finally examined using a Leica SP5II confocal microscope. The features sought were DNA fragmentation, condensation of the nuclei, changes in the outer membrane, or cytoskeletal abnormality. These features, if present, indicate cytotoxicity. RESULTS: The viability of the cultured nasal epithelial cells was unaltered by a 24-hour exposure to dexpanthenol, nor was the cellular proliferation rate affected at the level of statistical significance. There was evidence of a cytotoxic effect from exposing nasal epithelial cells to vaseline in liquid form for 24 hours. There was a reduction in cellular viability in the plates where the highest dose of vaseline (5 mg/mL) was used. Cellular viability was not affected significantly at any of the doses below 5 mg/mL. CONCLUSIONS: The absence of cytotoxic effects from the application of dexpanthenol to the nasal mucosa indicates that this agent may be safely used within the nose. The cytotoxic effects of liquid vaseline observed in this trial (condensed nuclear chromatin, loss of cellular volume) indicate that this agent may be harmful when used intranasally. For patients who require nasal packing due to nose bleeds or following endoscopic sinus surgical procedures, dexpanthenol should be preferred to vaseline from the point of view of maximizing healing of a nasal injury. [ABSTRACT FROM AUTHOR]

OBJECTIVE: In our study we aimed to evaluate the effects of applying propolis topically to epithelial cells of the nasal cells, to discover whether this causes any toxic effect upon the cells. MATERIALS AND METHODS: Samples of healthy human primary nasal epithelium harvested during septoplasty from volunteers were incubated in cell culture. MTT (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) assays may be utilized when assessing cellular damage (toxicity), as evidenced by DNA fragmentation, nuclear condensation, alteration in the outer plasma membrane and cytoskeletal alteration. This was the method used in the study. Cultured epithelial cells were incubated with propolis (Bee&You) for 24 hours at 37°C. The MTT assay was then performed, and the cell morphology was examined by confocal microscopy. In addition, via wound healing assay, cellular proliferation was assessed by the artificial scratch method followed by light microscopy. RESULTS: MTT assay results showed that the primary nasal cells were not affected from the topical application of propolis for 24 hours. All of the applied doses not changed significantly the viability of the cells. The agent was not found to cytotoxic to the primary nasal cells in the application time of 24 hours. Our confocal microscopy findings supported the MTT findings. According to the confocal images control cells that were not treated with test agent were with compact morphology and undamaged fusiform cell shape and nucleus. In test group of nasal cells, Propolis found not to be cytotoxic on the cellular morphology and not changed the cells. When evaluating the results from the wound healing assay, the clear area of scratch obtained at the start of incubation (0th) was closed totally with the proliferated primary nasal cells after incubation of 24 hours with propolis. These findings are supported by our MTT findings that imply to the slight induce of proliferation of the primary cells by Propolis. CONCLUSIONS: Topically applied propolis did not have a cytotoxic effect on nasal epithelium cells. Considering its antibacterial and antioxidant effects, it has been concluded that topical application in sinonasal inflammatory diseases (e.g., acute and chronic rhinosinusitis) may have an auxiliary effect in treatment. Moreover, there is a slight induce of proliferation of the primary cells by propolis which may help wound healing in septal surgeries and epistaxis. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The objective of this research was to examine the effects of topical ciprofloxacin on cultured nasal epithelial cells of human origin. MATERIALS AND METHODS: H uman n asal epithelial cells were collected from patients who voluntarily donated tissue left over following septorhinoplasty. The samples were from individuals without any indication of rhinosinusitis. An assay that may be employed to investigate toxic effects at the cellular level is MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). This test reveals where DNA becomes fragmented, the nuclei condense, the outer cell membrane is altered, or the cytoskeleton appears disrupted. The present study employed this technique. Nasal epithelium in cell culture was exposed to ciprofloxacin for 24 hours at a temperature of 37°C, following which the MTT assay was undertaken before examining the cells by confocal microscopy to look for alterations indicating cytotoxicity. Another test of toxicity, the artificial scratch technique, was also used. Cells treated in this way were assessed using a light microscope. RESULTS: The nasal epithelial cells in culture that were exposed to topical ciprofloxacin for 24 hours were less viable than controls and this result was statistically significant. For this length of exposure, the IC50 was calculated as 1.565 mg/mL. The peak reductions in cellular viability occurred with exposure at a concentration of 1.25 mg/mL and 0.625 mg/mL. These was only a mild decrease in viability at other concentrations, but these results were of statistical significance. The MTT assay and confocal microscopy confirmed this result. Cultured nasal epithelium not exposed to ciprofloxacin (i.e., controls) exhibited a compact morphological appearance when examined with confocal microscopy. The epithelial cells had a regular fusiform boundary, and the nuclei were intact. By contrast, the cultures with exposure to the antibiotic were of decreased size and their outline changed from fusiform to round. The epithelial cells cultures where scratch injury was induced were examined by light microscopy, the extent of closure of the denuded area being assessed after 24 hours. It was noted that the area opened up by the experimental scratch was not closed completely by 24 hours later. This result shows that ciprofloxacin decreases the viability of nasal epithelial cells. CONCLUSIONS: Topical application of cipro- floxacin to the nasal lining is not recommended, since this resulted in decreased cellular viability, cellular shrinking and alteration in outline from fusiform to round in cultured nasal epithelial cells. These changes indicate that topically applied ciprofloxacin is toxic to nasal epithelial cells. The outcomes of this study should be studied and correlated in vivo models. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The purpose of this study is to assess the effects of applying Garcinia cambogia to cultured human nasal epithelial cells. MATERIALS AND METHODS: A cell culture was set up consisting of human primary nasal epithelial cells harvested during septorhinoplasty from volunteers. The cells came from individuals with no history of rhinosinusitis. One assay for assessing cytotoxicity in cell culture utilises MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). This method allows visualization of fragmented DNA, condensation of nuclei and changes to the external cellular membrane or cytoskeleton. Our study employed this method. Nasal epithelial cells at 37°C were exposed in culture to G. cambogia for a period of 24 hours. Afterwards an MTT assay was used in conjunction with confocal microscopy to assess evidence of toxicity. The proliferative capability of the nasal epithelial cells was also evaluated by inducing a scratch injury to cultured cells followed by light microscopic examination. RESULTS: Testing for cytotoxicity in this manner indicates that G. cambogia does not appear harmful to cultured nasal epithelial cells when applied directly. The cells exposed to this plant extract were still fully viable 24 hours afterwards. There was no increase in viability at the level of statistical significance. It was noted, however, that proliferation did increase slightly within the exposure period. The MTT assay and confocal microscopy confirm these findings. Under confocal microscopic examination, a compact morphology with unaltered nuclear and cytoskeletal appearances was observed. Thus, there is no evidence suggesting viability is impaired or that cytotoxicity occurs. Ordinary light microscopic examination showed the area denuded of cells had become re-covered completely within 24 hours in the cultures where G. cambogia had been applied. The result suggests that exposure to G. cambogia has no significant effect in terms of stimulating or inhibiting cellular proliferation. CONCLUSIONS: G. cambogia may offer clinical benefit as a supplementary topical treatment for inflammation of the nose and sinuses, as seen in chronic and acute rhinosinusitis, or nasal polyps. The plant appears to increase nasal epitheliocytic proliferation slightly, as revealed by the MTT assay. There were no indications of a cytotoxic effect on epithelial cells of the nose. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The objective of the study was to evaluate the effect of ketoprofen when locally applied to tissue-cultured nasal epithelium. MATERIALS AND METHODS: Healthy primary nasal epithelial cells were grown in a tissue culture medium. MTT (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) assay was used to evaluate cytotoxicity. Markers of cellular injury revealed by the MTT assay include fragmentation of DNA, condensed nuclei, and changes affecting the cellular outer membrane and cytoskeleton. Epithelial cells at body temperature in cell culture were exposed over a 24-hour period to ketoprofen. Following the MTT assay, the confocal microscopic examination was performed. The extent to which epithelial cells remained capable of proliferating was evaluated by inducing a scratch injury, waiting for the repair to occur, and then examining the result with the ordinary light microscope. RESULTS: Topically applied ketoprofen does not affect the viability of tissue-cultured nasal epithelial cells within a 24-hour period. Furthermore, there were no cellular morphological alterations observed which would indicate toxicity from ketoprofen. In the scratch assay, the cells regained a normal confluent appearance within 24 hours. Thus, ketoprofen neither increases nor alters the rate at which nasal epithelial cells proliferate. CONCLUSIONS: Ketoprofen, when applied topically for 24 hours to nasal epithelial cells in cell culture, does not cause any alterations in cellular appearance which would suggest impairment of the ability to proliferate or indicate a cytotoxic effect. Extrapolating from these results, it appears acceptable to use ketoprofen topically within the nose in cases of rhinosinusitis (acute or chronic) or nasal pain since there is minimal risk of local toxic injury. [ABSTRACT FROM AUTHOR]