Nanocrocin Protective Effects on Paraquat-Induced Oxidative Stress in the MRC-5 Cell Line.

Paraquat (PQ) herbicide poisoning is a severe medical problem in developing countries without suitable therapy. This study aimed to investigate the effects of crocin (CCN) and nano crocin (NCCN) on PQ -induced toxicity in the MRC-5 cell line. The results showed that the particle size of NCCN was 140.3 ± 18.0 nm, and the zeta potential of the optimal crocin-loaded niosomes was 23.4 ± 2.8 mV. The NCCN was more effective than CCN in the inhibition of PQ-induced toxicity. Treatment of the MRC-5 cells leads to a decrease in ROS and an increase in SOD, CAT, GPX, and TAC levels in PQ-CCN and PQ-NCCN groups compared with the PQ group. These changes tended to be positively associated with the NCCN compared to CCN. Overall, NCCN was more effective than crocin in treating PQ-induced toxicity in vitro and deserved further preclinical consideration.

Crocin-loaded Niosomal Nanoparticles Reversing Cytotoxicity and Oxidative Stress in HEK293 Cell Line Exposed to Paraquat: An In vitro Study.

BACKGROUND: Paraquat (PQ) is an effective herbicide which is widely used around the world to remove weeds in agriculture. As a water-soluble carotenoid, crocin is a pharmacologically active constituent of C. sativus L. (saffron). OBJECTIVES: In the present study, we investigated the effects of crocin-loaded niosomes (Cro-NIO) compared to free crocin on PQ-induced toxicity in the eukaryotic human embryonic kidney (HEK293) cell line. METHODS: The Cro-NIO was synthesized and characterized. Cell viability was determined using the MTT assay in PQ-exposed HEK293 cell lines. The activities of biochemical markers were quantitatively determined to reveal the potential mechanism of PQ-induced oxidative stress in HEK293 cell line. RESULTS: The particle size, zeta potential, polydispersity index (PDI), DL, and EE of Cro-NIO were 145.4 ± 19.5 nm, -22.3 ± 3.11 mV, 0.3 ± 0.03, 1.74 ± 0.01%, and 55.3 ± 7.1%, respectively. PQtreated HEK293 cell lines decreased cell viability. The results of oxidative status showed that PQ significantly could increase ROS accumulation, accompanied by a decreasing antioxidant defense system. However, treatment with Cro-NIO, compared to crocin, not only did dose-dependently improve the cell viability but also significantly attenuated the ROS accumulation and increased antioxidant markers. CONCLUSION: According to these results, Cro-NIO, compared to crocin, was superior to ameliorating PQ-induced cytotoxicity and oxidative damage in HEK293 cells.

Cationic Dextran Stearate (Dex-St-GTMAC): Synthesis and Evaluation as Polymeric Micelles for Indomethacin Corneal Penetration.

Background Indomethacin as a non-steroidal anti-inflammatory drug (NSAID) is commonly used to treat some ocular inflammatory disorders. Unfortunately, indomethacin is a drug that is poorly soluble in water; therefore, it has low efficacy. An attractive approach is the targeted delivery of indomethacin to the cornea using cationic dextran stearate as a polymeric micelle drug carrier. Methods A dextran stearate-glycidyl trimethylammonium chloride (Dex-St-GTMAC) copolymer was prepared through the reaction of GTMAC, stearoyl chloride, and dextran. Then, Dex-St-GTMAC was characterized by Fourier transform infrared (FT-IR) spectroscopy and 1H NMR spectroscopy. Dex-St-GTMAC forms micelles in the presence of indomethacin. The prepared polymeric micelles were characterized for size, ζ-potential, drug loading, particle morphology, critical micelle concentration, and encapsulation efficiency. To study the irritation potential of the indomethacin-loaded Dex-St-GTMAC, Het-Cam and Draize tests have been performed. Prepared cationic micelles were subjected to the in vitro drug release and ex vivotrans-corneal permeation test. Results The dialysis method was used for the preparation of indomethacin-loaded micelles (10, 20, and 30%). Measurement of the particle size showed a mean diameter of 122.1 and 150.9 nm for the drug-loaded micelles. Scanning electron microscopy (SEM) images showed that the morphology of the particles is spherical. 10% formulation was chosen as the best formulation due to more surface charge and reasonable drug loading. ζ-potential measurement for the 10% drug-containing micelles showed a value of +39.1 mV. Drug loading efficiency and the encapsulation efficiency for 10% drug-containing micelles were 6.36 and 63.61%, respectively. The results of the Het-Cam and Draize tests indicated that the indomethacin-loaded Dex-St-GTMAC formulation had no toxicity to eye tissues. Based on our results, the prepared micelles (indomethacin-loaded Dex-St-GTMAC) exhibited a sustained drug release pattern compared to the control group. Indomethacin penetration from the micelles to the excised bovine cornea was 1.75-fold greater than the control (indomethacin 0.1% in phosphate-buffered saline (PBS)). Conclusions Data from the ζ-potential, SEM, drug loading capacity, and in vitro drug release studies indicated that cationic dextran stearate polymeric micelles are an appropriate carrier for the efficient penetration of indomethacin into cornea tissues.

Neuroprotective effects of crocin and crocin-loaded niosomes against the paraquat-induced oxidative brain damage in rats.

Paraquat (PQ) is a nonselective herbicide that induces oxidative reactions and multiple-organ failure on exposure. Crocin, a carotenoid obtained from saffron, has demonstrated many therapeutic effects against neural conditions because of its antioxidant properties. In this study, 30 male Wistar rats were divided into 6 groups to evaluate the protective effects of crocin and crocin-loaded niosomes (NC) against PQ in the brain. The levels of total antioxidant capacity (TAC), lipid peroxidation (LPO), total thiol groups (TTG), superoxide dismutase (SOD), and catalase (CAT) activity were measured as the markers of redox status. Histopathological changes in the CA1 region of the hippocampus were evaluated by cresyl violet staining. Results indicated that both crocin and NC were able to attenuate the adverse effects of PQ at the histopathological level, which was following the changes in LPO (P < 0.0001), TAC (P < 0.01), and TTG (P < 0.05) level. The activity of CAT (P < 0.01) and SOD (P < 0.01) could be restored either by crocin or NC. Also, results indicated that nanoformulation of crocin in niosomes appears to be more promising. In conclusion, both crocin and NC showed favourable effects of PQ in the brain of rats, and were determined to be excellent agents to prevent acute toxicities of PQ. Furthermore, these two compounds can be known to provide neuroprotection.

Protective Effects of N-acetylcysteine Niosome Nanoparticles on Paraquatinduced Nephrotoxicity in Male Rats.

INTRODUCTION: Paraquat (PQ), as a bipyridyl compound, is widely used as an effective herbicide that produces reactive oxygen species (ROS), affecting the unsaturated lipids of cell membranes leading to cell mortality. N-acetylcysteine (NAC) is a medication that has a beneficial role in reducing the intoxication of kidneys caused by PQ. Niosomes are bilayer vesicles that enhance the bioavailability of drugs. This study aimed to compare the effects of NAC and niosome of NAC (NACNPs) on PQ-induced kidney toxicity concerning its antioxidant activity. METHODS: In this experimental study, after formulating NACNP, 30 Wistar male rats weighing 180 to 250 gm were classified into five groups: the control group was treated with normal saline, while the other four groups received 35mg/kg/day of PQ via intraperitoneal route and, was treated with 25mg/kg/day NAC, 25mg/kg/day niosome and 25 mg/kg/day NACNP by gavage, Then, oxidative stress biomarkers such as total antioxidant capacity (TAC), catalase activity (CAT), lipid peroxidation (LPO), and total thiol group (TTG), plus blood urea nitrogen (BUN) and creatinine levels were evaluated in kidney tissue homogenate and examined histopathologically. RESULTS: The results revealed that TTG increased significantly in NAC & NACNP groups than in the PQ group. Further, in the PQ group, LPO increased significantly compared with the control, NAC, and NACNP groups, while in the NAC and NACNP group, LPO diminished compared with the PQ group. There was no significant difference in TAC between groups. Blood urea nitrogen (BUN) and creatinine levels dropped in NACNP compared with the PQ group and the NAC. Histological studies also approved PQ-induced damage and the protective effect of NACNP. CONCLUSION: The results indicated that NACNP could modulate oxidative stress status and kidney function against PQ toxicity.

Evaluation of Ascorbic Acid Niosomes as Potential Detoxifiers in Oxidative Stress-induced HEK-293 Cells by Arsenic Trioxide.

Background: As an environmental contaminant, Arsenic (As) poses many risks to human health. Increased Oxidative Stress (OS) and decreased antioxidant cell defense are the suggested mechanisms of carcinogenicity and toxicity of As. As a powerful antioxidant and water-soluble compound, vitamin C protects cells and tissues against oxidation and has a wide range of healing properties. Objectives: The current study aimed to formulate a suitable ascorbic acid (vitamin C) niosome and compare it with vitamin C in preventing As-induced toxicity in HEK-293 cells. Methods: Various formulas of vitamin C niosomes were prepared by C-SPAN mixed with cholesterol. The physicochemical characteristics of niosomal formulations, including load size, zeta-potential, and the drug release profile, were evaluated in HEK-293 cells. Then, OS biomarkers such as total reactive oxygen species (ROS), malondialdehyde (MDA), catalase (CAT), Antioxidant Capacity (TAC), and superoxide dismutase (SOD) activities determined the protective effects of vitamin C niosomes compared with vitamin C against As-induced toxicity. Results: The particle size and zeta potential of the optimal vitamin C niosome were 163.2 ± 6.1 nm and 23.3 ± 3.5 mV, respectively. Arsenic increased ROS and MDA levels while decreasing CAT, TAC, and SOD activities in the HEK-293 cell line. Finally, the vitamin C niosome decreased OS and increased antioxidant properties more than vitamin C. Significance: Vitamin C niosome was more effective than vitamin C in treating As-induced toxicity in vitro.

Adulteration of the Herbal Weight Loss Products by the Illegal Addition of Synthetic Antiobesity Medications: A Pilot Study.

BACKGROUND: Some anorexic agents are used to fraudulent augmentation herbal weight loss formulations. This study was designed to evaluate the potential existence of illicit substances in 63 herbal weight loss formulations collected from local apothecaries in Hamadan, Iran. METHODS: The thin-layer chromatography method was applied for the primary screening of potential illicit substances in the samples. The positive samples were analyzed using an isocratic high-performance liquid chromatography method. RESULTS: The results showed that 26.98% of the samples contained 17.76 ± 6.02 mg/cap of sibutramine. Daily therapeutic dose intake of sibutramine is in the range of 5 to 15 mg daily. CONCLUSION: Since apothecaries have advised consumers to take at least two capsules a day, it seems that the blood concentration of sibutramine will likely rise beyond the therapeutic concentration and become toxic. Therefore, the usage of such products could pose serious risks to consumers' health.

Beneficial Effects of Adding Topical Atorvastatin 5% Cream to Topical Betamethasone 1% Ointment on Chronic Hand Eczema.

BACKGROUND: Hand eczema (HE) refers to a common inflammatory dermatological condition. Several studies have shown that statins may have anti-inflammatory effects. This study aimed at investigating the efficacy of adding topical atorvastatin to topical betamethasone in the treatment of chronic HE. METHODS: This randomized, double-blind, placebo-controlled research was done between October 2017 and August 2018 in Hamadan, Iran. Of 130 cases treated for HE, 88 were randomly assigned to groups receiving either betamethasone 1% ointment plus atorvastatin 5% cream (n = 44) or betamethasone 1% ointment plus vehicle cream (n=44). Both groups applied their medications twice a day for 10 days. The primary outcome was changes in the severity of HE, assessed by hand eczema severity index (HECSI). The secondary outcomes were changes in itching evaluated via visual analogue scale (VAS) and quality of life examined through dermatology life quality index (DLQI). RESULTS: Seventy-two out of 88 eligible cases completed the study. The mean HECSI scores decreased in both groups after the intervention, although the change in HECSI was greater in the atorvastatin group (adjusted mean difference [AMD]: 5.756; 95% CI: 5.168 to 6.344, P<0.001). The mean VAS scores decreased in both groups after the intervention, although the change in VAS was greater in the atorvastatin group (AMD: 10.535; 95% CI: 7.005 to 14.064, P<0.001). Treatment with topical atorvastatin was more effective in improving DLQI (AMD: 1.990; 95% CI: 1.821 to 2.158, P<0.001). CONCLUSION: Addition of topical atorvastatin to topical betamethasone is beneficial in treatment of chronic HE. TRIAL REGISTERATION: Identifier: IRCT2017070922965N10; https://www.irct.ir/.

Improvement of therapeutic potential N-acetylcysteine in acetaminophen hepatotoxicity by encapsulation in PEGylated nano-niosomes.

AIMS: N-Acetylcysteine (NAC) is an effective antidote for the treatment of acetaminophen (APAP) poisoning; however, due to its low stability and bioavailability, repeated dosing of NAC is needed. This study investigated the therapeutic efficacy of NAC by niosomal carriers. MATERIALS AND METHODS: Niosomes were synthesized using surface active agents film hydration method and their physicochemical properties were characterized. In the in vivo study, in addition to control group, male rats were divided in different groups and challenged with an oral dose of APAP (2000 mg/kg); 4 h later, rats were administered normal saline, empty niosome (NIO), NAC (25 mg/kg) and NAC-loaded niosome (NAC-NIO) respectively, and sacrificed 48 h post-APAP overdose. KEY FINDINGS: The particle size and zeta potential of NAC-NIO were 242.3 ± 18.5 nm and -23.9 ± 1.6 mV. The loading and encapsulation efficiency of niosomes were 1.22% ± 0.02% and 26.76% ± 6.02%. APAP administration leads to hepatic damage as evidenced by increases in serum hepatic enzyme levels and tissue levels of nitric oxide and lipid peroxidation as well as decreases in hepatic levels of reduced glutathione, catalase, superoxide dismutase, and glutathione peroxidase. Treatment of rats with NIO-NAC was remarkably more effective than NAC in improving biochemical changes such as serum hepatic aminotransferases. These findings were correlated well to the histopathological experiments. SIGNIFICANCE: Our results suggest that NAC when delivered as a niosomal structure, is potentially more effective than NAC standard, in improving APAP-induced hepatotoxicity.

Local Anesthetic Effect of Amitriptyline versus Lidocaine in Isolated Lesion of the Limb Requiring Primary Suturing; Assessing a Novel Therapeutic Agent.

OBJECTIVE: To compare the anesthetic effects of topical amitriptyline 2% with lidocaine 2% in isolated limb wound repair with suturing. METHODS: In a randomized clinical trial, 90 patients with a complaint of isolated ulcer and require a preliminary repair by suturing were selected from patients referred to emergency department of Beast Hospital in Hamadan. First, the scars were washed and anesthetized with lidocaine 2%. If after the peak period effect of lidocaine, the pain score of patients did not decrease, they randomly assigned to two groups, Lidocaine or Amitriptyline gel. After the intervention and during the suturing, the patient's pain score was measured at the intervals specified time by the visual analogous scale (VAS) and results recorded on the checklist. Finally, the collected data were analyzed by SPSS software version 20 at 95% confidence level. RESULTS: In the lidocaine and amitriptyline group, the mean age of the patients was 29.08 and 27.34 years, and male gender frequency was 71.1% and 80% respectively. Both groups were matched for age and sex. Mean score of pain in both groups decreased from the score of 10 before the intervention to 7.33 in the lidocaine group and 0.53 in amitriptyline group. Based on the results of the ANOVA repeated measure test, there was a statistically significant difference between the mean score of pain in the two groups (p<0.001). CONCLUSION: In patients with isolated limbs ulcers, requiring initial repair with suturing, numbness and analgesia effect of amitriptyline 2% gel, with dose 2 mg/kg is better than lidocaine 2%.

Comparison of efficacy and safety of atorvastatin 5% lotion and betamethasone 0.1% lotion in the treatment of scalp seborrheic dermatitis.

Purpose: Seborrheic dermatitis (SD) is a chronic inflammatory skin disorder that mainly affects areas rich in sebaceous glands, such as the scalp. Although the exact cause of SD is not clearly understood, it seems that skin colonization with Malassezia fungus and the inflammatory responses of the immune system to this fungus play an important role in the pathology of SD. Recently a growing body of evidence has shown anti-inflammatory and anti-fungal effects of statins. Thus, this study aimed to evaluate the efficacy of topical atorvastatin in the treatment of scalp SD. Patients and methods: In this double-blind, clinical trial, 86 patients with mild-to-moderate scalp SD were divided into either atorvastatin (n=45) or betamethasone groups (n=41) by block randomization method. In addition to the ketoconazole 2% shampoo (3 times per week), the atorvastatin group received atorvastatin 5% lotion and the betamethasone group received betamethasone 0.1% lotion daily for 4 weeks. The SD severity of each patient was determined by Symptom Scale of Seborrheic Dermatitis (SSSD) at baseline and 4 weeks after treatment. Also, the patient's satisfaction of the treatment and adverse effects were investigated through individual reporting. Results: After 4 weeks of treatment, the score of SD severity decreased significantly in both groups, while changes of SSSD score from baseline to the fourth week of treatment were comparable in the two groups (P-value=0.476). Regarding patient's satisfaction of the treatment, results demonstrated the non-inferiority of atorvastatin as compared to betamethasone. Topical atorvastatin was also well-tolerated in almost all patients. Conclusion: Although preliminary, the results of the present study showed that topical atorvastatin has a comparable effect to topical betamethasone and can be considered as an alternative therapeutic modality in the treatment of scalp SD. However, these results need to be confirmed in future studies while taking into consideration the improvement of topical statin formulations.

The occurrence of acetaminophen/codeine as an adulterant in herbal analgesic supplements in Hamadan, Iran: A pilot study.

Analgesics, such as acetaminophen (APAP) and codeine (COD), are used to adulterate medicinal herbs and/or herbal supplements. This study evaluated the APAP and COD levels in 60 herbal supplement and/or herb-based medicine samples collected from apothecaries in Hamadan, Iran. The samples were analysed using a high-performance liquid chromatography (HPLC) system. The results showed that 15% of the samples contained 38900-165200 ng/g and 31.1-603.3 ng/g of APAP and COD, respectively. Due to the side-effects of analgesic drugs in human, control of these drugs is recommended in herbal supplements.

Role of Cerium Oxide Nanoparticles in a Paraquat-Induced Model of Oxidative Stress: Emergence of Neuroprotective Results in the Brain.

Paraquat (PQ), as a widely used herbicide, enhances the formation of free radicals and oxidative stress. Cerium oxide nanoparticles (CeNPs) are one of the most utilized and effective nanoparticles having strong antioxidative properties and inhibiting free radicals. Here, we aimed to investigate the effects of CeNPs on brain oxidative toxic stress injury induced with PQ. The male rats were treated intraperitoneally daily with PQ (50 mg/kg/day) and CeNPs (15, 30, and 60 mg/kg/day) alone or in combination for 2 weeks. After treatments, the brain tissue samples were collected. Oxidative toxic stress biomarkers including lipid peroxidation (LPO), total antioxidant capacity (TAC), and total thiol molecules (TTM) as well as DNA damage and caspase-3 levels were measured. Moreover, the mRNA expression levels of Nestin and Neurod1 were assayed. Our results showed that PQ has significantly increased brain LPO, DNA damage, and caspase-3 levels and further reduced TAC and TTM contents, as well as expression levels of Nestin and Neurod1, compared with the control group (injection of saline). CeNPs (15- and 30-mg/kg doses) in groups co-administered with PQ significantly ameliorated the LPO, DNA damage, and caspase-3 levels while increasing TAC and TTM contents as well as enhancing Nestin and Neurod1 mRNA expression levels in the brain samples (P < 0.05). These findings suggest a neuroprotective and antioxidant role for CeNPs in PQ-induced brain injury. However, further studies are required to clarify its clinical/pharmacological significance.

Cytotoxicity Enhancement of Paclitaxel by Loading on Stearate-g-dextran Micelles on Breast Cancer Cell Line MCF-7

Objective: Paclitaxel (PTX) is a chemotherapeutic agent used for treating breast cancer. The study aimed to prepare PTX loaded dextran stearate (Dex-SA) and evaluate its efficacy against human breast cancer cell line MCF-7. Methods: Dex-SA/PTX micelles were prepared by dialysis method. The micelles size, zeta potential and particle size distribution were measured by dynamic laser light scattering method. Amount of loaded PTX on the polymer measured by HPLC. Release profiles of the drug from the micelles were obtained in buffer (phosphate pH=7.4). Then the cytotoxicity of blank micelles, Dex-SA/PTX micelles and free PTX were evaluated in the MCF-7 cells by MTT method. Result: Loading efficiency of PTX on the Dex-SA was measured about 84.24±9.07%. The smallest particles size was about 193.9±7.1 nm but the other formulation with larger particle size had better zeta potential (-33.5±6.74 mV). The drug release from the micelles was slowly and reached steady state after about 12 hours. The cytotoxicity experiment showed that Dex-SA/PTX micelles have more cytotoxicity compared to free PTX against MCF7 cell lines. Conclusions: Dex-SA polymeric micelle is a suitable carrier for hydrophobic cytotoxic drugs such as PTX.

Uptake of etoposide in CT-26 cells of colorectal cancer using folate targeted dextran stearate polymeric micelles.

Targeted drug delivery using folate receptors is one of the most interesting chemotherapeutic research areas over the past few years. A novel folate targeted copolymer was synthesized using dextran stearate coupled to folic acid. FT-IR and NMR spectroscopy were used to confirm successful conjugation. Micelles prepared using this copolymer were characterized for their particle size, zeta potential, critical micelle concentration (CMC), drug loading capacity, and release efficiency. Cytotoxicity and cellular uptake of the micelles were estimated using CT-26 colorectal carcinoma cell line. FT-IR and NMR spectroscopy confirmed production of folate grafted dextran stearate copolymer. Low CMC value indicates that the copolymers are suitable for preparation of stable micelles useful in parenteral dosage forms. Particle size and zeta potential of the targeted nanoparticles were 105.5 ± 2.0 nm and -21.2 mV, respectively. IC50 of etoposide loaded in folate grafted dextran stearate enhanced about 20-fold compared to the pure drug (0.49 ± 0.11 µg/mL versus 9.41 ± 0.52 µg/mL). It seems that etoposide loaded in micelles of folate grafted dextran stearate copolymer is promising in reducing drug resistance of colorectal cancer by boosting etoposide cellular uptake.

Synthesis and in vitro evaluation of MR molecular imaging probes using J591 mAb-conjugated SPIONs for specific detection of prostate cancer.

Carcinoma of the prostate is the most frequent diagnosed malignant tumor in men and is the second leading cause of cancer-related death in this group. The cure rate of prostate cancer is highly dependent on the stage of disease at the diagnosis and early detection is key to designing effective treatment strategies. The objective of the present study is to make a specific MR imaging probe for targeted imaging of cancer cells. We take advantage of the fact that many types of prostate cancer cells express high levels of prostate-specific membrane antigen (PSMA) on their cell surface. The imaging strategy is to use superparamagnetic iron oxide nanoparticles (SPIONs), attached to an antibody (J591) that binds to the extracellular domain of PSMA, to specifically enhance the contrast of PSMA-expressing prostate cancer cells. Conjugation of mAb J591 to commercial SPIONs was achieved using a heterobifunctional linker, sulfo-SMCC. Two types of prostate cancer cell lines were chosen for experiments: LNCaP (PSMA+) and DU145 (PSMA-). MRI and cell uptake experiments demonstrated the high potential of the synthesized nanoprobe as a specific MRI contrast agent for detection of PSMA-expressing prostate cancer cells.