Agmatine prevents the memory impairment and the dysfunction of hippocampal GSK-3ß and ERK signaling induced by aluminum nanoparticle in mice.

The growing usage of aluminum nanoparticles (Al-NP) and their exposure may influence body function. Considering the proposed relationship between Al and the pathogenesis of Alzheimer's disease and the concern about the effect of this nanoparticle on brain health and cognitive function, the use of neuroprotective agents might be helpful. According to the reported neuroprotective effects of agmatine, in the present study, the possible protective effect of agmatine was assessed in mice model of Al-NP-induced memory impairment. In addition, due to the roles of hippocampal Glycogen synthase kinase-3 beta (GSK-3ß) and ERK signaling in memory and its disorders, these pathways were also investigated. Al-NP (10 mg/kg/p.o.) with/without agmatine (5 or 10 mg/kg/i.p.) was administered to adult male NMRI mice for 5 days. Novel object recognition (NOR) test session was used to assess cognitive function. Following the behavioral assessments, the hippocampi were used to determine the phosphorylated and total levels of GSK-3ß and ERK as well as GAPDH using western blot analysis. The results showed that Al-NP impaired NOR memory in mice while agmatine 10 mg/kg prevented the memory deficit induced by Al-NP. Furthermore, Al-NP activated GSK-3ß as well as ERK signals within the hippocampus while agmatine prevented the effects of Al-NP on GSK-3ß and ERK signals within the hippocampus. Besides supporting the neuroprotective effects of agmatine, these findings suggest the possibility of the connection of hippocampal GSK-3ß and ERK signaling in the neuroprotective effect of this polyamine against Al-NP.

A stereological study reveals nanoscale-alumina induces cognitive dysfunction in mice related to hippocampal structural changes.

Aluminum (Al) is known to induce neurotoxicity in both humans and rodents. Recent evidence has indicated that the toxicity of Al Oxide (Al2O3) nanoparticles (Al-NP), one of the most abundantly used engineered nanoparticles, is far greater than that of Al itself. To date, however, no information is available regarding the effect of Al-NP on the stereological parameters of hippocampus. In particular, no stereological studies have evaluated the effect of Al-NP on hippocampal CA1, dentate gyrus volume, and number of pyramidal and granular cells. Thus, the present study aimed to take a multidimensional approach to assess the concomitant cognitive, stereological, and apoptotic changes induced by a five-day Al-NP ingestion (10 mg/kg/day) in mice. The results demonstrated that the five-day Al-NP ingestion elicited a reduced preference to explore a novel object in the novel object recognition test (a hippocampal-dependent task). Perhaps contributing to this memory deficit, Al-NP induced additional alterations in the hippocampus of male NMRI mice in terms of (1) hippocampal volume (decreased the volume of the whole hippocampus, CA1, and dentate gyrus regions), (2) cell number (decreased the number of CA1 pyramidal neurons and dentate gyrus granular cells), and (3) increased cleaved caspase-3 in the whole hippocampus. These results provided new mechanistic insight to understand the impairing effect of AL-NP on the hippocampal function and structure.

A dose response effect of oral aluminum nanoparticle on novel object recognition memory, hippocampal caspase-3 and MAPKs signaling in mice.

The increasing use of aluminum nanoparticles (nano-Al) leads to increased human exposure and might affect human health. Considering the suggested connection between aluminum exposure and Alzheimer's disease (AD) pathogenesis, there is a concern about the effect of nano-Al on cognitive function and brain health. This study was aimed to assess the effect of a 5-day oral gavage of aluminum oxide nanoparticle (nano-Al) on memory and the phosphorylation levels of hippocampal p38, JNK (c-Jun N-terminal kinase), ERK (extracellular signal-regulated kinase) as well as cleaved caspase-3 in mice. Adult male NMRI mice were treated with nano-Al in doses 5 and 10 mg/kg/oral gavage for 5 days. The test session of novel object recognition (NOR) task was performed on day 5. Following the NOR test, the hippocampi were isolated for western blot analysis to determine the total and phosphorylated levels of p38, JNK, ERK as well as cleaved caspase-3 proteins. The results showed that nano-Al oral gavage in doses of 5 and 10 mg/kg impairs NOR memory in mice. Moreover, the memory impairing effect of nano-Al coincided with a dose dependent increase in phosphorylated p38 and cleaved caspase-3 in the hippocampus. It also increased the ratio of phosphorylated to total content of ERK in the hippocampus while JNK signaling was not affected by nano-Al. This study showed that nano-Al in doses as low as 5 and 10 mg/ kg ingested for 5 days impairs NOR memory and activates p38, ERK and cleaved caspase-3 in the hippocampus.

Evaluating the accuracy of simplified VCUG in comparison with traditional VCUG for diagnosis of urinary-reflux in children: Radiation dose reduction.

INTRODUCTION: Voiding cystourethrogram (VCUG) is an imaging technique for diagnosing urinary-reflux. VCUG requires X-ray exposure, increasing the risk of malignancies induced by ionizing radiation, especially in children. Considering the importance of early diagnosis of urinary-reflux using VCUG to prevent kidney complications and the significance of reducing the radiation dose during this imaging, this study intends to reduce the imaging views of VCUG and evaluate the urinary-reflux, using a reduced number of images. MATERIAL AND METHODS: Children referred to a University hospital who underwent VCUG participated in this study. The routine VCUG procedure contained five images. Two images of full-bladder and post-void of each patient were considered a simplified procedure and were evaluated by two radiologists to determine reflux and its grade. The sensitivity and specificity of simplified VCUG were determined. RESULTS: 182 patients with a mean age of 5.36±4.95 years old participated in this study. There were 90% and 82% agreement between routine and simplified VCUG results in the left and right kidneys. The simplified VCUG had 87.2% and 100% sensitivity and specificity in detecting the left-side reflux, 83 .9% and 100%, to detect the right-side reflux. CONCLUSION: The results showed that using two images instead of five can effectively diagnose and grade urinary-reflux. The simplified VCUG had a relatively good and acceptable agreement in comparison to the routine cystourethrogram. It can effectively reduce the patient's radiation dose, especially in patients who need to perform serial bladder imaging for follow-up after operation.

The effects of subchronic agmatine on passive avoidance memory, anxiety-like behavior and hippocampal Akt/GSK-3ß in mice.

Agmatine, a polyamine derived from l-arginine, has been suggested to modulate memory. However, the available evidence regarding the effect of agmatine on the memory of intact animals is contradictory. This study aimed to assess the dose-response effect of subchronic agmatine on passive avoidance memory and anxiety-like parameters of elevated plus maze in adult intact mice. Furthermore, considering the roles of Akt/GSK-3ß signaling pathway in memory and Alzheimer's disease, the hippocampal contents of phosphorylated and total forms of Akt and GSK-3ß proteins were determined using the western blot technique. Agmatine was administered intraperitoneally at the doses of 10, 20, 30, 40 and 80 mg/kg/daily to adult male NMRI mice for 10 days after which the behavioral assessments were performed. Upon completion of the passive avoidance test, the hippocampi were removed for western blot analysis to detect the phosphorylated and total levels of Akt and GSK-3ß proteins. Results showed the biphasic effect of agmatine on passive avoidance memory; in lower doses (10, 20 and 30 mg/kg), agmatine impaired memory whereas in higher ones (40 and 80 mg/kg) improved it. Though, agmatine in none of the doses affected animals' anxiety-like parameters in an elevated plus maze. Moreover, the memory-improving doses of agmatine augmented Akt/GSK-3ß pathway. This study showed the biphasic effect of agmatine on passive avoidance memory and an augmentation of hippocampal Akt/GSK-3ß signaling pathway following the memory-improving doses of this polyamine.

Preparation, Statistical Optimization and Characterization of Propolis-Loaded Solid Lipid Nanoparticles Using Box-Behnken Design.

Purpose: Propolis is a resinous material obtained by honeybees with many biological and pharmacological properties which can be used for treatment of various diseases. Current study aims to formulate and characterize propolis-loaded solid lipid nanoparticles (SLNs) carrier system. Methods: The prepared SLNs, composed of glyceryl monostearate (GMS), Soy lecithin, Tween 80 and polyethylene glycol 400 (PEG 400), were fabricated employing solvent emulsification-evaporation technique. In addition, the impact of several variables including concentration ratios of GMS/Soy lecithin and PEG 400/Tween 80 along with emulsification time were evaluated on the size, polydispersity index (PDI) and zeta potential of particles. SLN formulations were optimized using Box-Behnken design. The particles were freeze dried and morphologically studied by scanning electron microscopy (SEM). The in-vitro release profile of propolis entrapped in the optimized nanoparticles was investigated. Results: The mean particle size, PDI, zeta potential, entrapment efficiency (EE) and loading efficiency (LE) of optimized propolis-loaded SLNs were found to be 122.6±22.36 nm, 0.28±0.06, -26.18±3.3 mV, 73.57±0.86% and 3.29±0.27%, respectively. SEM images exhibited nanoparticles to be non-aggregated and in spherical shape. The in-vitro release study showed prolonged release of propolis from nanoparticles. Conclusion: The results implied that the proposed way of SLN preparation could be considered as a proper method for production of propolis loaded colloidal carrier system.

Effects of sub-chronic caffeine ingestion on memory and the hippocampal Akt, GSK-3ß and ERK signaling in mice.

Caffeine, one of the most widely consumed psychoactive substance in the world, has been shown to affect mood, memory, alertness, and cognitive performance. This study aimed to assess the effect of sub-chronic oral gavage of caffeine on memory and the phosphorylation levels of hippocampal Akt (protein kinase B), GSK-3ß (Glycogen Synthase Kinase-3beta) and ERK (extracellular signal-regulated kinase) in mice. Adult male NMRI mice were administered with caffeine at the doses of 0.25, 0.5, 0.75 and 1.5 mg/kg/oral gavage for 10 days before behavioral assessments. Upon completion of the behavioral tasks, the hippocampi were isolated for western blot analysis to detect the phosphorylated and total levels of Akt, GSK-3ß and ERK proteins. The results showed that sub-chronic caffeine ingestion at the dose of 0.5 mg/kg improves memory in mice both in passive avoidance and novel object recognition tasks. Furthermore, this memory enhancing dose of caffeine elevated the ratios of phosphorylated to total contents of hippocampal Akt, GSK-3ß and ERK. This study suggests that sub-chronic low dose of caffeine improves memory and increases the phosphorylation of hippocampal Akt, GSK-3ß and ERK proteins.

Nanomicellar curcuminoids attenuates renal ischemia/reperfusion injury in rat through prevention of apoptosis and downregulation of MAPKs pathways.

Renal ischemia/reperfusion (I/R) injury is considered as a main problem in clinical practice. Curcuminoids, the active constituents of turmeric, seem to have potential renoprotective effects. However, the poor bioavailability of curcuminoids restricts their therapeutic effects. In the present study, the effect of nanomicellar curcuminoids (NC) treatment on renal function, histology, total antioxidant capacity (TAC), total oxidative stress (TOS), caspase-3 level as well as mitogen activated protein kinases (MAPKs: JNK, p38 and ERK) phosphorylation were evaluated following renal I/R. Adult male Sprague-Dawley rats were administered NC at the dose of 25 mg/kg 1 h before renal ischemia induction. The animals were subjected to bilateral renal ischemia for 60 min and reperfusion for 24 h. Subsequently, blood urea nitrogen (BUN), creatinine (Cr), renal histopathology, TAC, TOS, and oxidative stress index, cleaved caspase-3 level, Bax and MAPKs signaling were evaluated. The results indicated that NC pretreatment at the dose of 25 mg/kg significantly improved renal function as well as histolopatholgical damages. Moreover, NC reduced the level of renal oxidative stress, cleaved caspase-3 and Bax (as the proapoptotic proteins) and suppressed the activated Jun N-terminal Kinase (JNK), p38 and extracellular receptor kinase (ERK) signaling induced by renal I/R. The findings of the current study indicate that NC might prevent the injury induced by renal I/R through suppression of oxidative stress, apoptosis and MAPKs pathways.

Pharmaceutical implants: classification, limitations and therapeutic applications.

Controlled/sustained delivery systems have been developed rapidly which show the ability to overcome the obstacles of traditional delivery systems. Daily development of biomedical and biomaterial sciences has brought more attention to the implantable delivery systems. As a result, these systems have found their position in the medical field since they were introduced. The advances in the polymeric science along with the other fields, make the production of a wide variety of implantable systems, possible. The influence of these systems in medical field could not be denied Here', the pharmaceutical applications which have been mostly focused on, are discussed. Since these systems have proven to be beneficial, researchers are trying to adjust their defects to the desired properties. Doing so, the path that implantable delivery systems have crossed so far should be studied, and that's the aim of this review. In the present report, the advantages of these systems in chemotherapeutic, contraceptive, neuropsychology, pain management, peptide delivery, ocular delivery, cardiovascular, orthopedic, and dental fields have been evaluated.

Fabrication of long-acting insulin formulation based on poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanoparticles: preparation, optimization, characterization, and in vitro evaluation.

The purpose of this research was the fabrication, statistical optimization, and in vitro characterization of insulin-loaded poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) nanoparticles (INS-PHBV-NPs). Nanopar-ticles were successfully developed by double emulsification solvent evaporation method. The NPs were characterized for particle size, entrapment efficiency (EE%), and polydispersity index (PDI). The NPs also were characterized by scanning electron microscopy (SEM), Fourier transformed infrared spectroscopy (FTIR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), and circular dichroism (CD). The optimum conditions were found to be 1.6% polyvinyl alcohol (PVA), 0.9% of PHBV, and 15 mg/ml of insulin with the aid of the Box-Behnken experimental design results. The optimized NPs showed spherical shape with particle size of 250.21 ± 11.37 nm, PDI of 0.12 ± 0.01, and with EE% of 90.12 ± 2.10%. In vitro drug release pattern followed Korsmeyer-Peppas model and exhibited an initial burst release of 19% with extended drug release of 63.2% from optimized NPs within 27 d. In conclusion, these results suggest that INS-PHBV-NPs could be a promising candidate for designing an injectable sustained release formulation for insulin.

Effect of acute caffeine administration on PTZ-induced seizure threshold in mice: Involvement of adenosine receptors and NO-cGMP signaling pathway.

PURPOSE: Caffeine is a non-selective antagonist of A1 and A2A adenosine receptors (ARs). In this regard, nitric oxide (NO) is partly involved in the central effects of caffeine. In this study, we examined the effect of acute caffeine administration on pentylenetetrazole (PTZ)-induced seizure threshold by focusing on A1Rs, A2ARs, and NO-cGMP signaling pathway. METHODS: NMRI male mice (25-30 g) received caffeine (5, 50, and 100 mg/kg) alone, whereas 8-CPT (1 and 5 mg/kg, a selective A1Rs antagonist), SCH-442416 (5 and 10 mg/kg, a selective A2ARs antagonist) or sildenafil (5 and 10 mg/kg, a phosphodiesterase 5 inhibitor) were administrated alone or as pre-treatment before caffeine. Seizure threshold was assessed by intravenous infusion of PTZ. Nitric oxide metabolites (NOx) were measured with the Griess method. RESULTS: When administrated alone, caffeine (5 and 50 mg/kg) and 8-CPT (1 and 5 mg/kg) significantly decreased seizure threshold, while 100 mg/kg of caffeine, SCH-442416 or sildenafil did not change it. Only pre-treatment with SCH-442416 (5 and 10 mg/kg) or sildenafil (5 and 10 mg/kg) before 100 mg/kg of caffeine significantly decreased seizure threshold. Moreover, NOx levels significantly decreased following alone administration of caffeine (100 mg/kg) or 8-CPT (5 mg/kg). CONCLUSION: The results of present study showed that 5 and 50 mg/kg of caffeine had a proconvulsant effect but caffeine at a dose of 100 mg/kg had no effect on seizure threshold. In addition, it seems that the effect caffeine on seizure threshold is partly mediated through ARs or modulation of the NO-cGMP signaling pathway.

Acetophenone benzoylhydrazones as antioxidant agents: Synthesis, in vitro evaluation and structure-activity relationship studies.

Acetophenone and its analogues are naturally-occurring compounds found in many foods and plants. In this study, a series of acetophenone benzoylhydrazones 5a-o were designed and synthesized as new potential antioxidant agents. Designed molecules contain hydrazone and phenolic hydroxyl moieties which possibly contribute to antioxidant activity. The antioxidant properties of compounds 5a-o in terms of reducing ability and radical-scavenging activity were assessed by using FRAP and DPPH tests, respectively. While the unsubstituted compound 5a had the superior capacity in the FRAP assay, the 2,4-dihydroxyacetophenone analogue 5g was the most potent radical scavenger in the DPPH method. The antioxidant potential of representative compounds 5a and 5g was further confirmed by TEAC and ORAC assays. Cell viability assays revealed that while the promising compounds 5a and 5g had no significant toxicity against HepG2 and NIH3T3 cells, they potently protected HepG2 cells against H2O2-induced oxidative damage at low concentrations. Furthermore, spectroscopic studies with different biometals demonstrated that 5g was able to interact with Cu2+ to form a 1:1 complex.

Development and characterization of electrosprayed nanoparticles for encapsulation of Curcumin.

Curcumin has been proven to be an effective herbal derived anti-inflammatory and antioxidant biocompatible agent. In this research, poly(lactic-co-glycolic acid) (PLGA) (as a biocompatible and generally recognized as safe (GRAS) polymer) nanoparticles containing Curcumin were electrosprayed from different polymeric solutions with different concentrations for the first time. Morphology of these nanoparticles in the absence/presence of Curcumin was evaluated by scanning electron microscope, transmission electron microscope, and X-ray photoelectron spectroscopy analyses. Perfectly shaped nanoparticles with an average size of 300 and 320 nm were observed for neat and Curcumin-loaded PLGA, respectively. Curcumin-loaded electrosprayed nanoparticles showed a normal moderate initial burst and then a prolonged release period. Weibull, Peppas, and modified Korsmeyer-Peppas models were applied to study the kinetic and mechanism of Curcumin release from PLGA nanoparticles. Results showed high specific surface area and spherical geometry of the nanoparticles. Effectiveness of the electrospray method as a promising technique for preparing Curcumin-loaded nanoparticles was confirmed in this study. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 285-292, 2018.

G-CSF loaded nanofiber/nanoparticle composite coated with collagen promotes wound healing in vivo.

Sustained release of functional growth factors can be considered as a beneficial methodology for wound healing. In this study, recombinant human granulocyte colony-stimulating factor (G-CSF)-loaded chitosan nanoparticles were incorporated in Poly(ε-caprolactone) (PCL) nanofibers, followed by surface coating with collagen type I. Physical and mechanical properties of the PCL nanofibers containing G-CSF loaded chitosan nanoparticles PCL/NP(G-CSF) and in vivo performance for wound healing were investigated. G-CSF structural stability was evaluated through SDS_PAGE, reversed phase (RP) HPLC and size-exclusion chromatography, as well as circular dichroism. Nanofiber/nanoparticle composite scaffold was demonstrated to have appropriate mechanical properties as a wound dresser and a sustained release of functional G-CSF. The PCL/NP(G-CSF) scaffold showed a suitable proliferation and well-adherent morphology of stem cells. In vivo study and histopathological evaluation outcome revealed that skin regeneration was dramatically accelerated under PCL/NP(G-CSF) as compared with control groups. Superior fibroblast maturation, enhanced collagen deposition and minimum inflammatory cells were also the beneficial properties of PCL/NP(G-CSF) over the commercial dressing. The synergistic effect of extracellular matrix-mimicking nanofibrous membrane and G-CSF could develop a suitable supportive substrate in order to extensive utilization for the healing of skin wounds. © 2017 Wiley Periodicals Inc. J Biomed Mater Res Part A: 105A: 2830-2842, 2017.

Effect of Prayer on Intensity of Migraine Headache: A Randomized Clinical Trial.

BACKGROUND AND AIM: Migraine is a common form of headache that affects patients quality of life negatively. In addition to pharmacologic treatment, there are a variety of nonpharmacologic treatments for migraine headache. In present study, we examined the effect of prayer on intensity of migraine pain. METHODS: In a prospective, randomized, controlled trial from October 2013 to June 2014, this study has been conducted in Kerman, Iran. We randomly assigned 92 patients in 2 groups to receive either 40 mg of propranolol twice a day for 2 month (group "A") or 40 mg of propranolol twice a day for 2 months with prayer (group "B"). At the beginning of study and 3 months after intervention, patients' pain was measured using the visual analogue scale. RESULTS: At the beginning of study and before intervention, the mean score of pain in patients in groups A and B were 5.7 ± 1.6 and 6.5 ± 1.9, respectively. According to results of independent t test, mean score of pain intensity at the beginning of study were similar between patients in 2 groups (P > .05). Three month after intervention, mean score of pain intensity decreased in patients in both groups. At this time, the mean scores of pain intensity were 5.4 ± 1.1 and 4.2 ± 2.3 in patients in groups A and B, respectively. This difference between groups was statistically significant (P < .001). CONCLUSIONS: The present study revealed that prayer can be used as a nonpharmacologic pain coping strategy in addition to pharmacologic intervention for this group of patients.

Effect of TNF-α Blockade in Gingival Crevicular Fluid on Periodontal Condition of Patients with Rheumatoid Arthritis.

BACKGROUND: Periodontitis and rheumatoid arthritis (RA) share a number of clinical and pathologic features, one of which is the presence of the tumor necrosis factor alpha (TNF-α)-induced bone resorption that is involved in the pathogenesis of both. OBJECTIVES: To investigate the effect of TNF-α blockade on periodontal conditions in patients with active RA. METHOD: The periodontal statuses of 36 patients (26 females, 10 males) diagnosed with active RA were evaluated both before and after anti-TNF-α therapy. Gingival index, bleeding on probing (BOP), probing pocket depth (PPD), oral hygiene index (OHI), and levels of TNF-α in gingival crevicular fluid (GCF) were measured at the baseline and 6 weeks after the treatment. Wilcoxon signed ranked test was used for statistical analyses. RESULTS: Based on OHI (p=0.860), the level of plaque control did not change during the study period, but there was a significant reduction in gingival inflammation based on the mean BOP (p=0.049) and GI (p=0.036) before and after 6 weeks of anti-TNF-α therapy. The mean PPD index did not significantly differ at the baseline and 6 weeks after treatment (p=0.126). CONCLUSION: Anti-TNF-α therapy might have a desirable effect on periodontal conditions and might reduce TNF-α level in GCF of patients with RA.

The frequency of using herbal medicines among patients with hypertension in Kerman, Iran, 2012-2013.

INTRODUCTION: The use of medicinal plants has dramatically increased in recent years. Given the increasing rate of hypertension and medical plants usage by these patients and considering drug interactions due to concomitant use with drugs, the present study aims to evaluate the rate of medicinal plants usage in hypertensive patients. METHODS: This is a cross-sectional study (descriptive-analytical) in which 650 hypertensive patients referring to the subspecialty clinic of Kerman were questioned about medicinal plants usage by a medicinal plants questionnaire. Among these patients, there were 612 who consented to participate. After the variables were described, the data were finally analyzed using Stata 12. RESULTS: The average age of those using these drugs in the past year was 58.8 ± 10 years. Of the total number of participants using medicinal plants, there were 58 males (23.5%) and 122 females (33.4%). There were 129 participants (72.5%) using medicinal plants through self-administration, 17 participants (9.5%) on experienced users' advice, 16 participants (9%) as administered by herbalists, and 11 participants (6%) as administered by doctors. However, the most important resources for using a drug that prevents hypertension were family and friends (74 participants; 41.5%) and doctors (13 participants; 7.3%). According to the results, there was no significant difference between the level of education and medicinal plants usage (P = .95); however, there was a significant difference between gender and medicinal plants usage (P = .009). DISCUSSION: According to the results indicating the relatively high prevalence of medicinal plants usage and their arbitrary use by hypertensive patients without consulting a specialist, it seems necessary to plan for more effective and secure public education and train people to provide herbal drug services for various diseases with hypertension being the most common one.

Transdermal absorption enhancing effect of the essential oil of Rosmarinus officinalis on percutaneous absorption of Na diclofenac from topical gel.

CONTEXT: Rosemary essential oil has been used topically for several purposes (analgesic, anti acne, and anti-inflammatory) in Iranian traditional medicine. OBJECTIVES: This investigation aimed to study the effect of essential oil of Rosmarinus officinalis L. (Lamiaceae) on the transdermal absorption of Na diclofenac from topical gel. MATERIAL AND METHODS: Diclofenac sodium topical gel was prepared with HPMC K4M and Carbopol 934P as a gelling agent, and several vehicles. The most stable gel was chosen and enhancing effects of the essential oil with different concentrations (0.1, 0.5, and 1.0% w/w) on the permeation of diclofenac were evaluated. The anti-nociceptive effect of preparations was evaluated based on the formalin and tail flick tests in mice. RESULTS: The major constituents of the essential oil were 1,8-cineol (15.96%), α-pinene (13.38%), camphor (7.87%), bornyl acetate (6.54%), verbenone (5.82%), borneol (5.23%), camphene (4.96%), and (E)-caryophyllene (3.8%). Topical diclofenac containing 0.5% essential oil showed more analgesic effect after 25, 30, and 35 min (p < 0.001) than the reference drug in the tail flick test. The analgesic effect of preparation containing 1% essential oil was more than reference gel after 15 min (p < 0.05). This difference was observed after 20, 25, 30, 35, and 40 min (p < 0.001) too. Rosemary essential oil 1% promoted analgesic effect of drug in comparison with diclofenac gel in the formalin early phase (p < 0.05). The enhancing effect of rosemary was observed in 0.5 and 1% concentration (p < 0.05 and p < 0.001, respectively) in the late phase. CONCLUSION: This study proved the enhancing effect of 0.5 and 1% of rosemary essential oil on diclofenac percutaneous absorption.