Chitosan-based emulgel and xerogel film containing Thymus pubescens essential oil as a potential wound dressing.

Controlling the wound exudates accompanied by microbial wound infections has still remained as one the most challenging clinical issues. Herein, a chitosan/gelatin/polyvinyl alcohol xerogel film containing Thymus pubescens essential oil is fabricated for antimicrobial wound dressing application. The chemical and physical characteristics of the devised formulation is characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, atomic force microscope, and tensile tests. Moreover, swelling capability, water vapour transmission rate, water contact angle, solubility, moisture content, and release properties are also studied. The antimicrobial and antibiofilm tests are performed using the broth microdilution and XTT assay, respectively. The produced formulation shows excellent antimicrobial efficacy against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida species. It is also demonstrated that the obtained film can reduce (∼80 %) Candida albicans biofilm formation, and its biocompatibility is confirmed with MTT (∼100 %) and hemolysis tests. The antimicrobial activity can be correlated to the microbial membrane attraction for Candida albicans cells, illustrated by flow cytometry. This proposed film with appropriate mechanical strength, high swelling capacity in different pH values (∼200-700 %), controlled release property, and antimicrobial and antioxidant activities as well as biocompatibility can be used as a promising candidate for antimicrobial wound dressing applications.

Encapsulation of Zataria multiflora essential oil in polyvinyl alcohol/chitosan/gelatin thermo-responsive hydrogel: Synthesis, physico-chemical properties, and biological investigations.

Zataria multiflora essential oil is a natural volatile plant product whose therapeutic applications require a delivery platform. Biomaterial-based hydrogels have been extensively used in biomedical applications, and they are promising platforms to encapsulate essential oils. Among different hydrogels, intelligent hydrogels have recently attracted many interests because of their response to environmental stimuli such as temperature. Herein, Zataria multiflora essential oil is encapsulated in a polyvinyl alcohol/chitosan/gelatin hydrogel as a positive thermo-responsive and antifungal platform. According to the optical microscopic image, the encapsulated spherical essential oil droplets reveal a mean size of 1.10 ± 0.64 µm, which are in consistent with the SEM imaging results. Encapsulation efficacy and loading capacity are 98.66 % and 12.98 %, respectively. These results confirm the successful efficient encapsulation of the Zataria multiflora essential oil within the hydrogel. The chemical compositions of the Zataria multiflora essential oil and the fabricated hydrogel are analyzed by gas chromatography-mass spectroscopy (GC-MS) and Fourier transform infrared (FTIR) techniques. It is found that thymol (44.30 %) and γ-terpinene (22.62 %) are the main constituents of the Zataria multiflora essential oil. The produced hydrogel inhibits the metabolic activity of Candida albicans biofilms (∼60-80 %), which can be related to the antifungal activity of the essential oil constituents and chitosan. Based on the rheological results, the produced thermo-responsive hydrogel shows a gel-sol viscoelastic transition at a temperature of 24.5 °C. This transition leads to a facile release of the loaded essential oil. The release test depicts that about 30 % of Zataria multiflora essential oil is released during the first 16 min. In addition, 2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay demonstrates that the designed thermo-sensitive formulation is biocompatible with high cell viability (over 96 %). The fabricated hydrogel can be deemed as a potential intelligent drug delivery platform for controlling cutaneous candidiasis due to antifungal effectiveness and less toxicity, which can be a promising alternative to traditional drug delivery systems.

Drug Delivery Assessment of a Novel Triple Antibiotic-Eluting Injectable Platelet-Rich Fibrin Scaffold: An In Vitro Study.

BACKGROUND: Intracanal disinfection is a critical, yet challenging goal for long-term success in regenerative-based treatments. This in-vitro study aimed to assess the release profile of triple antibiotic- eluting Injectable Platelet-Rich Fibrin (I-PRF) constructs in 28 days. METHODS: I-PRF scaffolds containing triple antibiotic mixture [Metronidazole (MET), Ciprofloxacin (CIP), and Minocycline (MINO)] by immersion (group one), I-PRF scaffolds containing triple antibiotic mixture by integration (group two), and antibiotic-free I-PRF scaffolds (group three) were fabricated. The antibiotic release from the scaffolds was measured using High-Performance Liquid Chromatography (HPLC) (the mobile phase of 0.1% formic acid and methanol (35:65 v/v), a C18 analytical column (150 × 4.6 mm, 5 µm) at a flow rate of 0.7 mL/min, at 25ºC) at days 1, 3, 7, 14, 21, and 28. RESULTS: Retention times for MINO, CIP, and MET were achieved as 2.3, 2.6, and 3.1 min, respectively. The maximum UV absorbance values for CIP, MET, and MINO were 268 nm, 278 nm, and 350 nm, respectively. The results of the first group showed burst release within the first 24 hours followed by sustained maintenance of all three antibiotics up to 14 days. MINO and MET were still detectable in the third week. The second group could not sustainably release the antibiotics. CONCLUSION: The developed method for the simultaneous identification and quantification of each antibiotic in I-PRF was sensitive and quick. Overall, group one could take up the antibiotics in adequate quantities and then subsequently release them over the study period.

Serum Tyrosine Level in Acute Murine Toxoplasmosis.

BACKGROUND: Toxoplasmosis is a zoonotic disease caused by the obligate intracellular parasite, Toxoplasma gondii. This global infectious disease has been associated with behavioral changes in rodents and can result in humans' neuropsychiatric symptoms. Since the neurotransmitters alteration can cause a behavioral change, in this study, tyrosine level, as a precursor of dopamine, was evaluated in acute murine toxoplasmosis during 2015 and 2016 in Shiraz, Iran. METHODS: At the first, 105 tachyzoites of T. gondii were subcutaneously inoculated to 50 BALB/c mice as experimental groups and 10 mice inoculated by PBS considered as the control group. After that, daily, one group of mice was bled, and sera were collected. Then, their serum tyrosine level was evaluated by HPLC method. RESULTS: After data analysis, the maximum mean serum tyrosine level was seen at 2th day of post parasite inoculation (0.0194 mg/ ml), with a significant difference compared to the control group (0.0117 mg/ ml, P=0.025). Moreover, the least quantity of serum tyrosine (0.076 mg/ml) was seen on the 5th day, after parasite inoculation, however, no significant difference was seen. CONCLUSION: Serum tyrosine level increased in 2 d after inoculation of Toxoplasma, but the level regularly decreased in successive days. Tyrosine level increased by phenylalanine hydroxylase 2 days after inoculation, then tyrosine decreased by tyrosine hydroxylase in the next days. Toxoplasma tyrosine hydroxylase enzymes, at primary days of toxoplasmosis, effect on tyrosine production, and after that, the most effect on tyrosine consumption.

Analytical methodologies for determination of methotrexate and its metabolites in pharmaceutical, biological and environmental samples.

Methotrexate (MTX) is a folate antagonist drug used for several diseases, such as cancers, various malignancies, rheumatoid arthritis (RA) and inflammatory bowel disease. Due to its structural features, including the presence of two carboxylic acid groups and its low native fluorescence, there are some challenges to develop analytical methods for its determination. MTX is metabolized to 7-hydroxymethotrexate (7-OH-MTX), 2,4-diamino-N10-methylpteroic acid (DAMPA), and the active MTX polyglutamates (MTXPGs) in the liver, intestine, and red blood cells (RBCs), respectively. Additionally, the drug has a narrow therapeutic range; hence, its therapeutic drug monitoring (TDM) is necessary to regulate the pharmacokinetics of the drug and to decrease the risk of toxicity. Due to environmental toxicity of MTX; its sensitive, fast and low cost determination in workplace environments is of great interest. A large number of methodologies including high performance liquid chromatography equipped with UV-visible, fluorescence, or electrochemical detection, liquid chromatography-mass spectroscopy, capillary electrophoresis, UV-visible spectrophotometry, and electrochemical methods have been developed for the quantitation of MTX and its metabolites in pharmaceutical, biological, and environmental samples. This paper will attempt to review several published methodologies and the instrumental conditions, which have been applied to measure MTX and its metabolites within the last decade.

Chemical Composition and Antimicrobial Activities of the Essential Oil From Salvia mirzayanii Leaves.

Resistance of many pathogens to available drugs is a global challenge and is leading to growing interest in natural alternative products. In this study, chemical composition and in vitro antibacterial and antifungal activities of the essential oil from Salvia mirzayanii were investigated. The chemical constituents of essential oil from S mirzayanii were analyzed by gas chromatography-mass spectrometry. The antimicrobial activity was determined by broth microdilution. The main identified compounds were 1,8-cineole (41.2 ± 1.3%), linalool acetate (11.0 ± 0.5%), and α-terpinyl acetate (6.0 ± 0.4%) (mL of essential oil/g of plant material). The MIC95 were 0.03 to 0.5 µL/mL and 16 to 128 µL/mL for gram-positive and gram-negative bacteria, respectively. These results indicated that Salvia mirzayanii essential oil significantly inhibited the growth of standard and clinically isolated tested yeasts by MIC50 0.03 to 1 µL/mL. Potent antibacterial and antifungal activities of Salvia mirzayanii essential oil may be considered in future study, particularly against antibiotic-resistant cases.

Chemical Composition and the Effect of Walnut Hydrosol on Glycemic Control of Patients With Type 1 Diabetes.

BACKGROUND: Walnut hydrosol (WH) is used extensively by Iranian people with diabetes in order to control blood sugar (BS). There are few data regarding the effect of walnut on controlling diabetes. OBJECTIVES: A pilot study to determine the efficacy and safety of WH in patients with type 1 diabetes. MATERIALS AND METHODS: Eight patients with diabetes mellitus (DM) type 1 were enrolled in the study. They did not use any medicine except insulin. They were advised to drink 250 mL WH after meals twice a day for four weeks. Their BS level was measured and their insulin dose was changed according to their BS. After four weeks, they discontinued WH use and their BS level was checked for two weeks. Descriptive statistics was used to analyze the data. Also, the essential oil of the sample was extracted using a liquid extractor and then analysis of the constituents was performed. RESULTS: The average daily BS level and insulin dose decreased in seven subjects. Two subjects developed generalized pruritic erythematous skin rash. One patient presented hypoglycemic coma. She had no other coma risk factor. Seven compounds were identified in the walnut essential oil and the rate of monoterpenoid and sesquiterpenes hydrocarbons were 53.45% and 5.95%, respectively. The main constituents of the oil were carvacrol (33.21%), thymol (16%) and homoveratrole (15.83%). CONCLUSIONS: WH may control the glycemic level in people with diabetes, but it may be associated with minor and major side effects. Further in vitro studies, using these seven compounds, are recommended to determine the efficacy and complications of WH in people with diabetes.