Encapsulation of eucalyptus essential oil in chitosan nanoparticles and its effect on MDA-MB-231 cells

Objective(s): Encapsulation of essential oil in polymeric nanoparticles (NPs) increases their retention and improves their efficacy. Here, eucalyptus essential oil (EEO) encapsulate in the chitosan (CS) NPs increases its retention, and enhances the anticancer effect of EEO.Materials and Methods: The effects of pH, chitosan sodium tripolyphosphate ratio, and chitosan concentration on the size and charge NPs were evaluated. The success of EEO encapsulation was confirmed by FT-IR, UV–Vis spectroscopy, and GC techniques. The toxic effect of free EEO and CS-EEO NPs was investigated in MDA-MB-231 breast cancer cells and fibroblast normal cells. Results: The optimized obtained EEO -loaded chitosan nanoparticles (CS-EEO NPs) were spherical with an average diameter of 86 nm, a polydispersity index below 0.4, and positive zeta potential (+14.25 mV) as confirmed. Increasing the concentration and pH of the chitosan solution and decreasing the chitosan/sodium tripolyphosphate ratio, the size of NPs decreased. Loading capacity (LC) and encapsulation efficiency (EE) of EEO in the NPs were about 45% and 32–76%, respectively. The chitosan nanoparticles exhibited a biphasic release profile with the release of 87% of the EEO in the first 5 h, followed by a sustained release for the next 43 h. Conclusion: The free EEO was more toxic for MDA-MB-231 cells than fibroblast cells; however, CS-EEO NPs were non-toxic for fibroblast cells and more toxic for MDA-MB-231 cells compared to free EEO. Therefore, the CS-EEO NPs illustrate smart behavior in killing cancerous cells and will be suggested for breast cancer drug delivery

Effects of Oral L-Carnitine Supplementation on Lipid Profile, Anemia, and Quality of Life in Chronic Renal Disease Patients under Hemodialysis: A Randomized, Double-Blinded, Placebo-Controlled Trial

In patients on maintenance hemodialysis several factors reduce the body stored carnitine which could lead to dyslipidemia, anemia, and general health in these patients. We evaluated the effect of oral L-carnitine supplementation on lipid profiles, anemia, and quality of life (QOL) in hemodialysis patients. In a randomized, double-blinded, placebo-controlled trial, end-stage renal disease (ESRD) patients on hemodialysis received either L-carnitine 1 g/d (n = 24) or placebo (27 patients) for 16 weeks. At the end of the study, there was a significant decrease in triglyceride (−31.1 ± 38.7 mg/dL, P = 0.001) and a significant increase in HDL (3.7 ± 2.8 mg/dL, P < 0.001) levels in the carnitine group. Decrease in total cholesterol (−6.6 ± 16.0 mg/dL, P = 0.075) and increase in hemoglobin (0.7 ± 1.7 g/dL, P = 0.081) concentrations in the carnitine group were not significant. There was no statistically significant changes in LDL in any group (P > 0.05). Erythropoietin dose was significantly decreased in both the carnitine (−4750 ± 5772 mg, P = 0.001) and the placebo group (−2000 ± 4296 mg, P < 0.05). No improvement was observed in QOL scores of two groups. In ESRD patients under maintenance hemodialysis, oral L-carnitine supplementation may reduce triglyceride and cholesterol and increase HDL and hemoglobin and subsequently reduce needed erythropoietin dose without effect on QOL

Comparison of antibacterial property of chitosan nanoparticles against Escherichia coli and Staphylococcus aureus

Background: With the advent of modern sciences such as nanotechnology, the hope for treatment of infectious diseases has increased. Nanochitosan is one of the most widely used nanomaterials in this field that has been considered due to its characteristics such as biocompatibility, nontoxicity and bactericidal activity. Objective: The aim of this study was to compare the antibacterial properties of chitosan nanoparticles against Escherichia coli and Staphylococcus aureus. Methods: This in vitro study was performed at Iranian Research Organization for Science and Technology in 2014. Chitosan nanoparticles were prepared based on the ionic gelation. The characteristics of the prepared nanoparticles were determined by DLS and SEM. The antibacterial activities of chitosan nanoparticles against Escherichia coli and Staphylococcus aureus were evaluated by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Data were analyzed using One-way ANOVA and T-test. Findings: Chitosan nanoparticles were formed with an average size of 160 nm. The MIC and MBC of chitosan nanoparticles were 0.25 and 1 mg/ml for Escherichia coli and were 0.5 and 2 mg/ml for Staphylococcus aureus. The diameter of zones of inhibition was 19 mm for Escherichia coli and 14 mm for Staphylococcus aureus in 10 mg/ml concentration of chitosan nanoparticles. Conclusion: With regards to the results, it seems that nanochitosan has acceptable antibacterial activity against Escherichia coli and Staphylococcus aureus. But Escherichia coli is more sensitive to chitosan nanoparticles than Staphylococcus aureus. Keywords: Nanotechnology, Chitosan, Anti-Bacterial Agents, Escherichia Coli, Staphylococcus Aureu

Synergistic Cytotoxic Effect of Gold Nanoparticles and 5-Aminolevulinic Acid-Mediated Photodynamic Therapy against Skin Cancer Cells

Background: Photodynamic therapy (PDT) is a promising therapeutic modality for the treatment of cancer and other diseases. In this study, the epidermoid carcinoma cell line A431 and the normal fibroblasts were used to investigate whether gold nanoparticles (GNPs) can induce an increase in cell death during PDT using 5-aminolevulinic acid (5-ALA) as a photosensitizer. Methods: Human fibroblast and A431 cells were grown in 96-well plates. The effect of GNPs on the efficacy of 5-ALA-mediated PDT (5-ALA-PDT) was evaluated by comparing the effect of 5-ALA with GNPs to the effect of 5-ALA alone. Cell viability was determined by the methyl- tetrazolium assay. Results: Dark toxicity experiments showed that 5-ALA at concentrations 0.5, 1 and 2 mM had no significant effect on cell viability of both cell lines. However, treatment of cells with 5-ALA (0.5 to 2 mM) and light dose of 25 Jcm-2 led to 5-10% and 31-42% decrease in cell viability of fibroblast and A431 cells respectively. The data also shows that GNPs in both the absence and the presence of light, results in a dose-dependent decrease in cell viability of both cell lines. However, the sensitivity of cancer cells to GNPs at concentrations more than 24 μg/ml was approximately 2.5- to 4-fold greater than healthy cells. Furthermore, data indicates that 5-ALA in combination with GNPs results in a synergistic reduction in viability of A431 cells. Conclusion: In summary, the findings of this study suggest that concomitant treatment with 5-ALA and GNPs may be useful in enhancing the effect of 5-ALA-PDT

Cytotoxic effects of different solvents and essential oil of eucalyptus on human fibroblast cells

Background: There are reports that essential oils of different species of eucalyptus have special properties including anti-bacterial, antiviral, anticancer and insecticidal activities. Most of plant essential oils are dissolved in polar or non-polar solvents before examination. Objective: The aim of this study was to determine the cytotoxic effects of different solvents and essential oil of eucalyptus on human fibroblast cells. Methods: This experimental study was performed in the Institute of Biotechnology affiliated to Iranian Research Organization for Science and Technology during 2013-2014. The studied solvents were including ethanol, propylene glycol, dichloromethane, and dimethyl sulfoxide. Fibroblast cells were seeded in 96-well plates. After 24 h incubation at 37ºC, the cells were treated with different concentrations of the solvents or essential oil of eucalyptus (0.05-1 mg/ml). The viability of the cells was determined by the colorimetric MTT assay. Data were analyzed using T-test and One-way ANOVA. Findings: The lowest cytotoxic effect against fibroblast cells was related to 1% ethanol and propylene glycol, while the highest cytotoxic effect was related to 10% dimethyl sulfoxide that decreased cell survival by 47%. Essential oil of eucalyptus also induced fibroblast cell death at concentrations higher than 0.05 mg/ml. Conclusion: For clinical application of essential oil of eucalyptus, using solvent with appropriate concentration is necessary to prevent its cytotoxic effects. Keywords: Eucalyptus, Solvents, Fibroblast

The Effect of Nanoformulation of Amoxicillin on its Antibacterial Activity Against Common Bacterial Strains Involved in Hospital-Acquired Infections

Abstract Background: Nanotechnology offers a great chance to treat drug-resistant microbial infections. The purpose of this study was to synthesize amoxicillin-encapsulated polymeric nanoparticles and compare the antibacterial activity of this nanoformulation with pure amoxicillin. Materials and Methods: Amoxicillin-encapsulated polymeric nanoparticles were prepared using chitosan as a polymer and alginate as cross linking agent. The obtained nanoparticles were characterized by Fourier transform infrared, dynamic light scattering and scanning electron microscopy. The antibacterial effects of the nanoparticles were evaluated using broth microdilution and well diffusion methods against some common bacterial strains involved in hospital-acquired infections. Results: The drug-encapsulated nanoparticles were found to be spherical in shape with average size of 96 nm. These nanoparticles had a significant antibacterial effect on all tested bacteria, except for Pseudomonas aeruginosa. They also displayed stronger antibacterial activity than the nanocarrier alone or free antibiotic. The highest mean zones of growth inhibition (23.7 mm) for methicillin-sensitive Staphylococcus aureus and the smallest zone (12.5 mm) for the resistant species of this bacterium were determined. MIC of the nanoformulation against these two strains was respectively determined at 1.5 and 48 µg/ml and for Escherichia coli and Pseudomonas aeruginosa at 6.6 and 256 µg/ml. Conclusion: The results suggest that amoxicillin encapsulation in polymer nanoparticles has the potential to increase its antibacterial activity against bacteria causing nosocomial infections

Comparison of Antimicrobial Activity of Chitosan Nanoparticles Loaded with Amoxicillin with Free Amoxicillin against Some Gram Positive and Gram Negative Bacteria

Background and Objectives: Antibiotic resistance has prepared the way for substituting new therapeutic methods. Studies have indicated that nanoformulated antimicrobial agents have better therapeutic effects. In this study, the antimicrobial activity of chitosan nanoparticles loaded with amoxicillin (ACNs), was assessed in comparison with free amoxicillin against&nbsp;some&nbsp;Gram-positive and Gram-negative bacteria. &nbsp; Methods: In this experimental study, the nanoparticles were prepared using the ionotropic gelation technique. The resulting nanoparticles were characterized by dynamic light scattering (DLS), scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. The antibacterial activity of amoxicillin and nanoparticles against standard and clinical strains of methicillin-susceptible and methicillin-resistant Staphylococcus aureus, Escherichia coli, and Enterococcus faecalis, was investigated by determination of minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and inhibition zone diameters. Data were analyzed using one-way ANOVA with Tukeyʼs post hoc test. &nbsp; Results: The size of chitosan nanoparticles (CNs) and ACNs was 88 and 106 nm, respectively. ACNs showed higher antibacterial activity compared to amoxicillin and CNs without drug as determined by the&nbsp;smaller&nbsp;MIC (0.375mg/ml)&nbsp;and MBC (2 mg/ml) values and larger zones of inhibition (28mm). The highest and the lowest sensitivity to nanoparticles, were observed for Enterococcus feacalis and methicillin-resistant Staphylococcus aureus, respectively.&nbsp; &nbsp; Conclusion: The results of this study showed that the nanoformulation of amoxicillin may be an appropriate strategy to increase its therapeutic effects. However, precise clinical studies are required for its confirmation

In vitro and in silico studies of novel synthetic ACE-inhibitory peptides derived from Saccharomyces cerevisiae protein hydrolysate

The structure-function relation of YR-10 (YGKPVAVPAR) was investigated by synthesizing four structural analogs of that including YHR-10 (YGKHVAVHAR), GA-8 (GKPVAVPA), GHA-8 (GKHVAVHA), and PAR-3 (PAR). GA-8 (GKPVAVPA) was synthesized on the basis of simulated enzymatic gastrointestinal digestion performed by bioinformatics tools (expasy-peptide cutter). This study explains the molecular mechanisms for the interaction of synthetic peptides with ACE. The IC50 values of each were 139.554 +/- 2.3, 61.91 +/- 1.2, 463.230 +/- 3.56, 135.135 +/- 2.1, 514.024 +/- 5.86 mu M, respectively. Results indicated that Pro replacement with His in YR-10 and GA-8 increased ACE inhibitory activity respectively, by 55.63% and 70.82%. Removal of Tyr and Arg from respectively N and C terminal positions of YR-10, following in silico simulated gastrointestinal digestion caused the 3.31 fold decrease in ACE inhibitory activity. YHR-10 showed the best docking poses, and GHA-8 exhibited interaction with Zn2+. Lineweaver-Burk plots of most active peptides suggest that they act as noncompetitive inhibitors against ACE

Synthesis, in vitro and cellular antioxidant activity evaluation of novel peptides derived from Saccharomyces cerevisiae protein hydrolysate : structure-function relationship

The relationship between structure and function of primary antioxidant peptide, YR-10 (YGKPVAVPAR) was considered by synthesizing three analogues including YHR-10 (YGKHVAVHAR), GA-8 (GKPVAVPA) and PAR-3 (PAR). Antioxidant activity was determined through in vitro and cellular assays. Substitution of Pro with His in the structure of YR-10 led to significant (P < 0.05) higher ABTS radical scavenging and ferric reducing activity. Following in silico simulated gastrointestinal digestion, Tyr and Arg were omitted, respectively, from N and C-terminal positions and resulted in decreasing DPPH, ABTS radical scavenging, and ferric reducing activities. PAR-3 showed the best inhibitory activity on linoleic acid oxidation. Pretreatment of Caco-2 cells with YR-10, YHR-10, and GA-8 (1000 mu M) before exposure to H2O2 (160 mu M) resulted in 34.10%, 39.66% and 29.159% reduction in malondialdehyde and 53.52%, 17.02% and 24.71% reduction in protein carbonyl levels. The peptide pretreatment reduced catalase level in cells and PAR-3 exhibited the most protective effects on the viability of cells exposed to oxidative stress