Growth and invasion inhibition of T47D ductal carcinoma cells by the association of docetaxel with a bioactive agent in neutral nanosuspension.

Introduction: The approach for drug delivery has impressively developed with the emergence of nanosuspension, particularly the targeted nanoemulsions (NEs). It can potentially improve the bioavailability of drugs, enhancing their therapeutic efficiency. This study aims to examine the potential role of NE as a delivery system for the combination of docetaxel (DTX), a microtubule-targeting agent, and thymoquinone (TQ) in the treatment of human ductal carcinoma cells T47D. Methods: NEs were synthesized by ultra-sonication and characterized physically by dynamic light scattering (DLS). A sulforhodamine B assay was performed to evaluate cytotoxicity, and a flow cytometry analysis for cell cycle, apoptosis, autophagy, and cancer stem cell evaluations. A quantitative polymerase chain reaction further assessed the epithelial-mesenchymal transition gene expirations of SNAIL-1, ZEB-1, and TWIST-1. Results: The optimal sizes of blank-NEs and NE-DTX+TQ were found at 117.3 ± 8 nm and 373 ± 6.8 nm, respectively. The synergistic effect of the NE-DTX+TQ formulation significantly inhibited the in vitro proliferation of T47D cells. It caused a significant increase in apoptosis, accompanied by the stimulation of autophagy. Moreover, this formulation arrested T47D cells at the G2/M phase, promoted the reduction of the breast cancer stem cell (BCSC) population, and repressed the expression of TWIST-1 and ZEB-1. Conclusion: Co-delivery of NE-DTX+TQ may probably inhibit the proliferation of T47D via the induction of apoptosis and autophagy pathways and impede the migration by reducing the BCSC population and downregulating TWIST-1 expression to decrease the epithelial-to-mesenchymal transition (EMT) of breast cancer cells. Therefore, the study suggests the NE-DTX+TQ formula as a potential approach to inhibit breast cancer growth and metastasis.

Antitumour effects of a solid lipid nanoparticle loaded with gemcitabine and oxaliplatin on the viability, apoptosis, autophagy, and Hsp90 of ovarian cancer cells.

The present study aimed to explore the sensitising capability of the anticancer agents, gemcitabine (GEM) and oxaliplatin (OXA), encapsulated in a novel SLN (GEM:OXA-SLN) against the ovarian cancer cell lines. A novel SLN, prepared using hot homogenisation by mixing phosphatidylcholine, cholesterol, tween 80, and oleic acid, was characterised using Transmission Electron Microscope and zetasizer. The anticancer activities and the underlying molecular mechanisms of GEM:OXA-SLN were investigated. The average z-diameter of the homogeneous spherical GEM:OXA-SLN was (70.33 ± 0.70) nm with zeta potential (-7.69 ± 0.61) mV. GEM:OXA-SLN significantly inhibited the viability of ovarian cancer cells in a dose-dependent manner within 24 h. It also triggered the induction of autophagy cellular death, suppression of multidrug resistance efflux pump and inhibition of heat shock protein (Hsp90). The encapsulation of GEM and OXA in SLN improved the efficacy of the drugs and diminished the ovarian cancer cell's resistance.

Incorporation of docetaxel and thymoquinone in borage nanoemulsion potentiates their antineoplastic activity in breast cancer cells.

Combining more than one anticancer agent in a nanocarrier is beneficial in producing a formula with a low dose and limited adverse side effects. The current study aimed to formulate docetaxel (DTX) and thymoquinone (TQ) in borage oil-based nanoemulsion (B-NE) and evaluate its potential in impeding the growth of breast cancer cells. The formulated B-NE and the combination (DTX + TQ) B-NE were prepared by the ultra-sonication method and physically characterized by the dynamic light scattering techniques. The cytotoxicity analyses of (DTX + TQ) B-NE in MCF-7 and MDA-MB-231 cells were evaluated in vitro by using the SRB assay. Cell death mechanisms were investigated in terms of apoptosis and autophagy pathways by flow cytometry. The optimum mean droplet sizes formulated for blank B-NE and the (DTX + TQ) B-NE were 56.04 ± 4.00 nm and 235.00 ± 10.00 nm, respectively. The determined values of the half-maximal inhibitory concentration (IC50) of mixing one-half amounts of DTX and TQ in B-NE were 1.15 ± 0.097 µM and 0.47 ± 0.091 µM in MCF-7 and MDA-MB-231 cells, respectively, which were similar to the IC50 values of the full amount of free DTX in both tested cell lines. The treatment with (DTX + TQ) B-NE resulted in a synergistic effect on both tested cells. (DTX + TQ) B-NE induced apoptosis that was integrated with the stimulation of autophagy. The produced formulation enhances the DTX efficacy against human breast cancer cells by reducing its effective dose, and thus it could have the potential to minimize the associated toxicity.

Carrying epirubicin on nanoemulsion containing algae and cinnamon oils augments its apoptotic and anti-invasion effects on human colon cancer cells.

The nanotherapeutics holds great potential in cancer therapy since they may consist of more than one anticancer agent that has a different mechanism of action. The present study aimed to incorporate the epirubicin (EPI) into a nanoemulsion containing the algae and cinnamon oils (ALG-CN-EPI) using ultrasonication technique. The apoptotic efficacy of ALG-CN-EPI was assessed in the HCT116 human colon cancer cells using the assays of CCK-8, DNA fragmentation, reactive oxygen species (ROS) generation, and Annexin V-FITC/PI while the anti-invasion effect of ALG-CN-EPI was determined by the transwell invasion assay. The zeta average diameters and zeta potential of the nano-suspensions of ALG-CN-EPI, measured by the zetasizer, were 117.2 ± 3.02 nm and -1.810 ± 0.07 mV, respectively. Results of the apoptotic evaluation revealed that the half-maximal inhibitory concentration (IC50) of ALG-CN-EPI (0.7 ± 0.21 µM) was distinctly lower than that of free EPI (6.00 ± 1.56 µM). The DNA fragmentation of HCT116 cells was amplified by a factor of 8 ± 0.24 when treated with ALG-CN-EPI but it did not considerably differ when treated with the free EPI (1.13 ± 0.31). Additionally, cells treated with ALG-CN-EPI resulted in a significant elevation of the intracellular ROS production and higher percentages of late apoptotic cells relative to the EPI treated cells. ALG-CN-EPI treatment suppressed the invasion ability of HCT116 cells to (32.98 ± 3.28)%, whereas the invasion ability of EPI exposed cells was only reduced to about (56 ± 1.81)%. In conclusion, the resulted new nanotherapeutics (ALG-CN-EPI) has potentiated the antitumor activity of EPI.

Incorporation of methotrexate into coconut oil nanoemulsion potentiates its antiproliferation activity and attenuates its oxidative stress.

Methotrexate (MTX), a chemotherapeutic agent, has limited clinical applications due to its pulmonary and neurotoxicity. The antineoplastic activity of MTX-NE COCO, which is MTX formulated in coconut oil nanoemulsion (NE), was evaluated in A549 non-small cell lung cancer cells while its adverse side effects on the oxidative stress of the lung and brain were assessed in mice. The z-average diameter for the dispersed nanodroplet of MTX-NE COCO (79.74 ± 3.49 nm) was considerably greater than the free-NE COCO (64.80 ± 3.34 nm). In contrast, the magnitude of the negative z-potential of MTX-NE COCO (3.00 ± 0.69 mV) was markedly less than that of free-NE COCO (8.20 ± 0.76 mV). The minimum inhibitory concentration (IC50) of MTX-NE COCO (18 ± 1.8 µM) was less than the IC50 of free MTX (32 ± 1.2 µM) by around twofold. The in vivo evaluation of the MTX-NE COCO treatment revealed that the antioxidant enzymes activities of the brain and lung tissues, catalase, superoxide dismutase, and glutathione reductase, were relatively raised while the malondialdehyde amount was diminished when compared to the free MTX treatment. In conclusion, combining MTX with coconut oil in a NE had improved its efficacy while ameliorating its oxidative stress effect on the brain and lungs.

Incorporating ifosfamide into salvia oil-based nanoemulsion diminishes its nephrotoxicity in mice inoculated with tumor.

Introduction: Nephrotoxicity is one of the major side effects of the chemotherapeutic drug, ifosfamide (IFO). In this study, IFO was solubilized in nanoemulsion (NE) containing salvia (SAL) essential oil to investigate its adverse side effects in mice. Methods: One hundred female Swiss albino mice (n = 20/group) were split into five groups. Group I (Normal) received saline solution (0.9% (w/v) NaCl) while groups II-V were intraperitoneally (I.P.) injected with 2.5 × 106 Ehrlich ascetic carcinoma (EAC) cells/mouse. Group II (EAC) represented the untreated EAC-bearing mice. Group III (IFO) was treated with IFO at a dose of 60 mg/kg/d (I.P. 0.3 mL/mouse). Group IV (SAL) was treated with 0.3 mL blank NE-based SAL oil/mouse. Group V (SAL-IFO) was treated with IFO, loaded in 0.3 mL of blank SAL-NE, at a dose of 60 mg/kg/d (I.P. 0.3 mL/mouse). Groups III-V were treated for three consecutive days. Results: There was a double increase in the survival percentage of the SAL-IFO group (60%) relative to the IFO group (30%). Renal damage with the presence of Fanconi syndrome was indicated in the IFO group through a significant elevation in the levels of serum creatinine, blood urea nitrogen, urine bicarbonate, and phosphate in addition to a reduced level of glucose compared to the normal group. On the other hand, the administration of SAL-IFO into the mice reversed this effect. Additionally, the oxidative stress in the kidney tissues of the SAL-IFO group was ameliorated when compared to the IFO group. Conclusion: Incorporating IFO into SAL-NE has protected the kidneys from the damage induced by IFO.

Evaluation of Antitumor Activity and Hepatoprotective Effect of Mitomycin C Solubilized in Chamomile Oil Nanoemulsion.

PURPOSE: The present study aimed to investigate the antitumor activity and hepatoprotective effect of the MTC, when combined with CHAM oil nanoemulsion (NE), (CHAM-MTC) on the tumor growth. MATERIALS/METHODS: The in vitro study assessed the antineoplastic effect of CHAM-MTC on the MCF-7 breast cancer cells while the in vivo therapeutic effectiveness and toxicities of CHAM-MTC were evaluated in Ehrlich Ascites Carcinoma (EAC) bearing mice. One hundred female Swiss albino mice, divided equally into non-EAC group (negative control), untreated EAC group (positive control) and three EAC groups received once intraperitoneal injection of 0.2ml CHAM-NE, 0.2ml Normal Saline (NS) contained MTC (1mg/kg) and 0.2ml CHAM-NE mixed with MTC (1mg/kg), respectively. RESULTS: The in vitro results indicated that CHAM-NE could potentiate the effect of MTC in sub-effective concentrations since the half-maximal inhibitory concentration (IC50) was reduced by a factor of 21.94 when compared to the MTC-NS. The in vivo study revealed that mice treated with CHAM-MTC showed a significant increase in the median survival time (MST= 37 days) when compared to the MTC-NS treated group (MST= 29.50 days). In addition, CHAM-MTC showed protective ability against the oxidative stress and hepatic damage induced by EAC and MTC treatment. CONCLUSION: The combination of MTC with CHAM-NE could be valuable in enhancing the therapeutic efficacy of MTC against EAC and in eliminating MTC-induced hepatotoxicity.

Enhanced antitumour activity of doxorubicin and simvastatin combination loaded nanoemulsion treatment against a Swiss albino mouse model of Ehrlich ascites carcinoma.

Doxorubicin (DOX) is the most commonly used anticancer drug; however, it has limited use because prolonged administration may result in severe cardiotoxicity. Simvastatin (SIM), generally prescribed for hypercholesterolaemia, has also shown salubrious results in the monotherapy or combinational drug therapy of different cancers in various models. Nanoparticle drug delivery systems are a novel way of improving therapeutics and also improving the absorption and specificity of drugs towards tumour cells. In this study, we exploited this technology to increase drug specificity and minimize imminent adverse effects. In this study, the antitumour activity of the combination formulas of DOX and SIM, either loaded in water (DOX-SIM-Solution) or nanoemulsions (NEs) (DOX-SIM-NE), was evaluated in a Swiss albino mouse model of Ehrlich ascites carcinoma. The anticancer effect was assessed by quantifying the change in body weight, mean survival time, and percent increase in lifespan (%ILS), determining haematological and serum biochemical parameters (liver function test, kidney function test and lipid profile parameters) as well as studying the histopathological alterations in liver tissues. We observed a clear increase in %ILS of the DOX-SIM-Solution group (265.30) that was double the %ILS of the DOX-SIM-NE group (134.70). However, DOX-SIM-NE had a non-toxic effect on the haematological parameters, whereas DOX-SIM-Solution increased the levels of haemoglobin and lymphocytes. Furthermore, the encapsulation of SIM and DOX into NEs improved the levels of all serum biochemical parameters compared to the DOX-SIM-Solution. A reduction in the side effects of DOX-SIM-NE on the liver was also established using light microscopy, which revealed that the morphologies of the hepatocytes of the mice were less affected by administration of the DOX-SIM-NE treatment than with the DOX-SIM-Solution treatment. The study showed that incorporating SIM into the DOX-loaded-NE formulation remarkably improved its efficiency and simultaneously reduced its adverse effects.

Incorporation of ifosfamide into various essential oils -based nanoemulsions ameliorates its apoptotic effect in the cancers cells.

The chemotherapeutic drugs, loaded in nanocarriers, have recently attracted the pharmaceutical industries due to their limited adverse side effects. The objective of the current study was to incorporate the ifosfamide (IFO) into two different essential oils-based nanoemulsions, lemon (LEM-IFO) and salvia (SAL-IFO). The antiproliferation activities of the resulted formulas were evaluated in the MCF-7 breast cancer cells and HeLa cervical cancers cells. The cytotoxic effect of the NE formulas was detected by the MTT assay, DAPI stain and light microscopy. The z-average diameters range of LEM-IFO and SAL-IFO, determined by the zetasizer, were 49.15-61.81 nm and 56.64-64.62 nm, respectively. The half maximal inhibitory concentration (IC50) of LEM-IFO and SAL-IFO, applied into the HeLa cells, were 0.165 ± 0.025 and 0.141 ± 0.035 mM, respectively, whereas the IC50 of LEM-IFO and SAL-IFO subjected into the MCF-7 cells were 0.200 ± 0.005 mM and 0.270 ± 0.025 mM, respectively. The IC50 of the free IFO was markedly larger than LEM-IFO and SAL-IFO when applied into MCF-7 cells (9.20 ± 2.01 mM) and HeLa cells (7.69 ± 1.88 mM). Among the tested formulas, LEM-IFO and SAL-IFO have the greatest apoptotic effect on the MCF-7 and HeLa cells, respectively. Solubilizing the IFO in the essential oils-based NE has ameliorated the antitumor efficacy of IFO.

Cytotoxicity and apoptosis enhancement in breast and cervical cancer cells upon coadministration of mitomycin C and essential oils in nanoemulsion formulations.

The present study aimed to solubilize the antineoplastic agent, mitomycin C (MMC), in two nanoemulsions (NEs) consisting of different essential oils (ginger (Gi) and frankincense (Fr)) in order to examine their anticancer activities on the HeLa cervical cancer cells and MCF-7 breast cancer cells. The two NEs-based Gi and Fr oil were produced by a high-pressure homogenization technique followed by solubilizing of the MMC in both NE formulas. The produced formulas were physically characterized by zetasizer and were applied on HeLa and MCF-7 cells at various concentrations for 24 h. The cytotoxicity assays were performed in vitro, using MTT assay, Coomassie blue staining for cellular morphology evaluation, and DAPI fluorescent staining for molecular cell death assessment. The average droplet diameters of the blank NEs have markedly increased and the charges of the droplets were significantly reversed when MMC was loaded. The potential cytotoxicity of the blank and combined formulas on HeLa and MCF-7 cells were dose-dependent and significantly greater than the toxicities of the free MMC. Among the MMC-loaded NE formulas, Fr-MMC has endured the nuclear apoptosis in HeLa cells at a lower concentration and reported the least % of florescence uptake compared to Gi-MMC. In contrast, the combination formula, Gi-MMC, has the strongest apoptotic effect on the MCF-7 cell line since it has the least % florescence uptake compared to the other formulations. Mixing MMC with Gi-NE and Fr-NE has considerably improved its cytotoxicity on the MCF-7 and HeLa cells.

Antineoplastic activity of mitomycin C formulated in nanoemulsions-based essential oils on HeLa cervical cancer cells.

Combining the essential oils (ESSOs) with the chemotherapeutic agent, mitomycin C (MMC), in nanoparticle can be beneficial in cancer therapy. The aim of the current study was to in vitro evaluate the antineoplastic effect of MMC, formulated in two different nanoemulsions (NE) based on two ESSOs, chamomile (Ch) and garlic (Gar), on HeLa cervical cancer cells. The z-average diameter of Ch-NE has slightly increased from 83.39 ±â€¯12.85 nm to 91.18 ±â€¯5.79 nm when mixed with MMC (Ch-MMC) whereas the z-average diameter of Gar-NE has markedly increased from 50.6 ±â€¯1.96 nm to 75.64 ±â€¯7.13 nm when loaded with MMC (Gar-MMC). The zeta potentials of both of Ch-NE and Ch-MMC, which were -1.91 ±â€¯4.38 mV and -5.44 ±â€¯5.26 mV, respectively, have differed from Gar-NE and Gar-MMC, which were 11.4 ±â€¯2.29 mV and 11.5 ±â€¯2.28 mV, respectively. Compared to MMC solution, the cell viabilities of HeLa cells, measured by the MTT assay, were reduced 42 and 20 times when subjected into Ch-MMC and Gar-MMC, respectively. The light microscopy images revealed that the cell membrane of the HeLa cells treated with Gar-NE or Gar-MMC were more altered relative to the cells treated with Ch-NE or Ch-MMC. In contrast, the nuclei of the HeLa cells, stained with DAPI and treated with Ch-NE or Ch-MMC, were more fragmented than the cells treated with Gar-NE or Gar-MMC, indicating that both of Ch-NE and Ch-MMC have passed the cell membrane and affected the nucleus directly whereas Gar-NE and Gar-MMC have got attached to the cell membrane causing damage to the cell. In conclusion, combining MMC with NE-based ESSOs has increased the cytotoxic effect of the MMC on the HeLa cells with different mechanism of actions.

In Vivo Evaluation of the Anticancer Activity of the Gemcitabine and Doxorubicin Combined in a Nanoemulsion.

CONTEXT: Doxorubicin (DOX) and gemcitabine (GEM) are anticancer drugs that were combined in a nanoemulsion (NE) to reduce their adverse side effects. AIM: To detect the antitumor activity of the combination formulas of GEM and DOX, loaded either in water (GEM+DOX-Sol) or in NEs (GEM-DOX combination/loaded NE [GEM+DOX/LNE]), in female Swiss albino mice inoculated with Ehrlich ascites carcinoma (EAC). SETTINGS AND DESIGN: The anticancer assessment of the NE formulas was implemented in 200 mice, which were divided into 10 groups. MATERIALS AND METHODS: It includes the detection of the change in body weight, analysis of the hematological and serum biochemical profiles, and study of the histopathologic alterations of the heart tissues. STATISTICAL ANALYSIS: One-factor analysis of variance was used. RESULTS: Mice treated with GEM + DOX/LNE, which have an z-average of 155.38±2.33nm and zeta potential of -38.5±1.3 mV, recorded a considerable improvement in the mean survival time (MST), which was 60 days, as compared to the EAC control group, which has an MST of 28 days. It also restored the hematological and serum biochemical parameters toward normal values. CONCLUSIONS: The combination of GEM and DOX in NE has significantly diminished the cardiotoxicity of DOX and hematotoxicity of GEM while improving their antitumor properties.

Antimicrobial Activity of Biocompatible Microemulsions Against Aspergillus niger and Herpes Simplex Virus Type 2.

BACKGROUND: Microemulsions (MEs), which consist of oil, water, surfactants, and cosurfactants, have recently generated considerable interest as antimicrobial agents. OBJECTIVES: To determine the antifungal and antiviral activities of three ME formulations (MEa, MEb, and MEc) that differ in their hydrophilicity. METHODS: The ME formulas were produced by mixing different fractions of Tween 80, Span 20, ethanol, oil, isopropyl myristate, and distilled water. The antifungal activity of the ME formulas against Aspergillus niger, A. flavus, Bacillus, Candida albicans, and C. glabrata were determined by the solid medium diffusion cytotoxicity test against the mitochondria, measuring the minimum inhibitory concentration, dry biomass, and leakage of potassium, and characterizing the cell morphology. The antiviral activities of the ME formulas against the herpes simplex virus type 2 (HSV-2) were determined using the cytopathic effect assay. RESULTS: Significant antimicrobial activities were recorded against A. niger and herpes simplex virus type 2 (HSV-2) when treated with MEb that had hydrophobic nanodroplets with an average diameter of 4.7 ± 1.22 nm. A volume of 0.1 mL of MEb (10 mL of potato dextrose broth) inhibited the germination of A. niger cells, reduced their dry biomass, enhanced the leakage of potassium from the cell membranes, affected their mitochondria, and altered the shape of their conidia, in addition to enlarging them. MEb was able to destroy the HSV-2 virus at a 200-fold dilution in Dulbecco's modified eagle medium. CONCLUSIONS: The water-in-oil ME with equivalent surfactant-to-oil ratio (MEb) has great potential as an antifungal and antiviral agent.