Keratinocyte Exosomes for Topical Delivery of Tofacitinib in Treatment of Psoriasis: an In Vitro/ In Vivo Study in Animal Model of Psoriasis.

INTRODUCTION: Exosomes are extracellular vesicles in the range of 40-150 nm released from the cell membrane. Exosomes secreted by keratinocytes can communicate with other keratinocytes and immune cells with specific biomarkers at their surface, which may be effective on inflammation of psoriasis and its pathogenesis. OBJECTIVE: The present study aimed to formulate and study effectiveness of an exosomal delivery system of tofacitinib (TFC). METHODS: TFC was loaded by different methods in exosomes and then characterized for particle size, zeta potential, drug loading efficiency, and release efficiency. By comparing these parameters, the probe sonication method was chosen to load TFC into exosomes. The MTT assay was used to compare the cytotoxicity of the free drug with the TFC-loaded exosomes (TFC-Exo), and Real-time PCR was used to determine the expression levels of several genes involved in psoriasis expressed in the A-431 keratinocyte and their suppression after treatment. Animal model of psoriasis was induced in BALB/c mice by imiquimod and the efficacy of free TFC, and TFC-Exo were studies on macroscopic appearance and histopathological symptoms. RESULTS: Exosomes encapsulating TFC showed lower cytotoxicity in MTT assay, higher suppression the expression of TNF-a, IL-23, IL-6, and IL-15 genes in real-time PCR and better therapeutic effect on animal models compered to free TFC. CONCLUSIONS: This method of drug delivery for TFC may be effective on enhancing its therapeutic effects and reduction its side effects favorably in chronic administration.

Ocular delivery of sunitinib-loaded nanoparticles doped in tragacanthic acid hydrogel in treatment of diabetic retinopathy in rats.

OBJECTIVE: Diabetic retinopathy (DR) is a common microvascular complication of diabetes mellitus. This study aimed to compare the effect of sunitinib-loaded poly (glycerol sebacate) (PGS)/gelatin nanoparticles doped in an injectable hydrogel with bevacizumab as a standard treatment of DR. METHODS: The shear-sensitive hydrogel was prepared based on tragacanthic acid (TA) cross-linked with sodium acetate. DR was induced in rats by streptozotocin (STZ), and the animals were injected intravitreally a single dose of 20 µL sunitinib solution in three different concentrations (12.5, 25, and 50 µg/mL), sunitinib-loaded nanoparticles in hydrogel (413 µg/mL) and bevacizumab solution (6.25 mg/mL). The efficacy of the treatments was studied by histological and immunohisitological tests, angiogenesis, and optical coherence tomography (OCT). Vascular endothelial growth factor (VEGF) concentration was measured in the retina. RESULTS: The results revealed that 20 µL of sunitinib with the concentration of 25 µg/mL was effective in DR without any disruption in the retina or any other side effects. This dose was considered the therapeutic dose for nanoparticles. Sunitinib loaded PGS/gelatin nanoparticles that were incorporated in the injectable hydrogel were as effective as bevacizumab in controlling DR. Although sunitinib solution reduced VEGF production and neovascularization in the retina compared to the negative control group, it was not as suitable as the nanoparticles. TA-based hydrogel showed no toxicity on the normal retina, and the angiography and histologic studies confirmed the VEGF results.' CONCLUSIONS: Sunitinib nanoparticles doped in TA hydrogel may be an appropriate substitution of bevacizumab in the treatment of DR.

Preparation and characterization of a hydroxypropyl methylcellulose based wafer for simultaneous delivery of phenytoin and insulin as wound dressing material.

In this study, a novel wafer based on Hydroxypropyl methylcellulose (HPMC) was prepared as a wound dressing for the simultaneous delivery of phenytoin (PT) and insulin; evaluation of the cutaneous wound repair property was performed too. Due to its low water solubility, PT was encapsulated in polymeric micelles (PM) by the film hydration method at different polymer/drug ratios and characterized in terms of particle size (PS), polydispersity index (PdI), zeta potential (ZP), drug loading (DL) %, entrapment efficiency (EE) %, and drug release. Then, the optimized PT loaded PM (PT-PM) was embedded in the wafers prepared from the HPMC polymer, alone or in combination with Carbopol 940 (CB) and xanthan gum (XG). This wafer also contained a fixed amount of insulin (PT-PM-Insulin-wafer). The obtained wafers were evaluated in terms of morphology, water uptake ability, porosity, bioadhesion and hardness features. Finally, the efficacy of the PT-PM-Insulin-wafer was assessed in full-thickness excision wound models. The optimized PT-PM showed the PS of 84.05 ± 1.80 nm, PdI of 0.28 ± 0.22, ZP of -3.38 ± 0.26 mV, DL of 15.63 ± 0.01%, EE of 92.66 ± 0.08%, and the release efficiency of 59.95 ± 0.03%. The results obtained from the XRD studies of PT-PM also demonstrated the transition of the crystalline nature of the PT to the amorphous form, while FTIR studies showed some intermolecular interaction of PT and the Soluplus® copolymer chain. It was also found that the incorporation of XG into HPMC wafers influenced the microstructure, thus increasing the porosity, water uptake ability and bioadhesion. Compared with other groups, the PT-PM-Insulin-wafer group showed the enhancement of wound closure through increasing collagen deposition and re-epithelialization. The present study, therefore, revealed that the PT-PM-Insulin-wafer group might have very promising applications for wound healing.

Sustained-release of erythropoietin using a novel injectable thermosensitive hydrogel: in vitro studies, biological activity, and efficacy in rats.

In the current study erythropoietin (EPO) loaded trimethyl chitosan/tripolyphosphate nanoparticles-embedded in a thermosensitive hydrogel was prepared. The influence of the main experimental factors on the properties of EPO-loaded nanoparticles were evaluated using a two-factors central composite design and the optimized formulation was then freeze dried. Sodium dodecyl sulfate-page and circular dichroismspectroscopy were used to confirm the structural stability of EPO following encapsulation and freeze drying. Rheological properties, and the release rate of EPO from the hydrogel were examined. Mean particle size, zeta potential, and entrapment efficiency of the optimized EPO-loaded nanoparticles were confirmed 151.5 ± 16 nm, 11.5 ± 1.8 mV, and 78.5 ± 5.9%, respectively. The hydrogel containing nanoparticles existed as a solution at room temperature converted to a semisolid upon increasing the temperature to 35 ± 1.2 °C and demonstrated controlled release of EPO for more than 10 days. The stability of EPO in the hydrogel system was further investigated using in vivo biological activity assay and the result revealed relative potency of 0.85 as calibrated with standard EPO. Finally, a single injection of the EPO-loaded nanoparticles-embedded in the hydrogel administered to Sprague-Dawley rats resulted in elevated reticulocytes for about 20 days compared to control group received blank hydrogel.

Novel pH-triggered biocompatible polymeric micelles based on heparin-α-tocopherol conjugate for intracellular delivery of docetaxel in breast cancer.

In this study, pH-triggered polymeric micelle comprising α-tocopherol (TOC) and heparin (HEP) was developed and loaded with docetaxel (DTX). The amphiphilic copolymer was synthesized by grafting TOC onto HEP backbone by a pH-cleavable bond. DTX-loaded micelles were characterized in terms of critical micelle concentration (CMC), particle size, zeta potential, entrapment efficiency (EE), pH-responsive behavior, and drug release. In vitro cytotoxicity of the micelles against breast cancer cells was investigated by MTT assay. The cellular uptake of coumarin-loaded micelles was also evaluated. Furthermore, the pharmacokinetics of DTX-loaded micelles was evaluated and compared with that of Taxotere®.HEP-CA-TOC copolymers showed low CMC values and high EE. At pH 7.4, the micelles remained stable in size and shape, whereas considerable changes in particle size and morphology were observed at pH 5.5. DTX-loaded micelles showed pH-dependent drug release profiles. Coumarin-loaded micelles showed higher cellular uptake than free coumarin. Therefore, the DTX-loaded micelles showed more toxicity against breast cancer cells than free DTX. A significant increase in T1/2 ß, AUC0-∞ and MRT was observed in DTX-loaded micelle treated group as compared to the group treated with Taxotere®.The results suggest that the pH-sensitive HEP-modified micelles could be promising for enhanced intracellular drug delivery of DTX for cancer treatment.

Enhanced solubility, oral bioavailability and anti-osteoporotic effects of raloxifene HCl in ovariectomized rats by Igepal CO-890 nanomicelles.

The purpose of the present study was to enhance the bioavailability and anti-osteoporotic effects of raloxifene HCl (RH) by increasing its solubility and inhibition of the p-glycoprotein pump using surfactant micelles of Igepal CO-890. The micelles were prepared by the direct method and their critical micellar concentration, drug dissolution rate, saturated solubility, drug loading and surface morphology were defined. The cytotoxicity of Igepal CO-890 and its ability to inhibit the p-glycoprotein pump were studied on Caco-2 cells. The pharmacokinetic parameters were analyzed by oral administration of a single dose of 15 mg/kg in Wistar rats. Anti-osteoporotic effects were studied by measuring the calcium, phosphorous, and uterus weight of rats after one month of oral administration of 6 mg/kg/day of RH in ovariectomized rats. Igepal CO-890 micelles enhanced the RH solubility by about two-fold. The FT-IR and DSC studies indicated no interaction between the drug and the surfactant. XRD spectrum showed an amorphous state of RH in the micelles. The p-glycoprotein pump was inhibited by Igepal CO-890 in Caco-2 cells comparable to verapamil. Micelles increased the uterine weight and decreased the serum calcium and phosphorus significantly compared to the untreated drug. Oral bioavailability of RH increased about four-fold by nanomicelles.

Development and in vitro/in vivo evaluation of HPMC/chitosan gel containing simvastatin loaded self-assembled nanomicelles as a potent wound healing agent.

The aim of this study was to develop hydroxypropyl methyl cellulose (HPMC)/chitosan gel containing polymeric micelles loaded with simvastatin (Sim) and evaluates its wound healing properties in rats. An irregular full factorial design was employed to evaluate the effects of various formulation variables including polymer/drug ratio, hydration temperature, hydration time, and organic solvent type on the physicochemical characteristics of pluronic F127-cholesterol nanomicelles prepared using the film hydration method. Among single studied factors, solvent type had the most impact on the amount of drug loading and zeta potential. Particle size and release efficiency was more affected by hydration temperature. The optimized formulation suggested by desirability of 93.5% was prepared using 1 mg of Sim, 10 mg of copolymer, dichloromethane as the organic solvent, hydration time of 45 min and hydration temperature of 25 °C. The release of the drug from nanomicelles was found to be biphasic and showed a rapid release in the first stage followed by a sustained release for 96 h. The gel-contained nanomicelles exhibited pseudo-plastic flow and more sustained drug release profile compared to nanomicelles. In excision wound model on normal rats, the wound closure of the group treated by Sim loaded micelles-gel was superior to other groups. Taken together, Sim loaded micelles-gel may represent a novel topical formulation for wound healing.

Synthesis, in vitro characterization, and anti-tumor effects of novel polystyrene-poly(amide-ether-ester-imide) co-polymeric micelles for delivery of docetaxel in breast cancer in Balb/C mice.

OBJECTIVE: The goal of the present work was to make novel co-polymeric micellar carriers for the delivery of docetaxel (DTX). SIGNIFICANCE: Co-polymeric micelles can not only solubilize DTX and eliminate the need for toxic surfactants to dissolve it, but also cause passive targeting of the drug to the tumor and reduce its toxic side effects. METHODS: Poly(styrene-maleic acid) (SMA) was conjugated to poly (amide-ether-ester-imide)-poly ethylene glycol (PAEEI-PEG). Copolymer synthesis was proven by Fourier transform infrared (FTIR) and 1H-nuclear magnetic resonance (1H-NMR). The SMA-PAEEI-PEG micelles loaded with DTX were prepared and their critical micelle concentration (CMC), zeta potential, particle size, entrapment efficiency, and their release efficiency were studied. MCF-7 and MDA-MB231 breast cancer cells were used to evaluate the cellular uptake and cytotoxicity of the micelles. The antitumor activity of the DTX-loaded nanomicelles was measured in Balb/c mice. RESULTS: The FTIR and HNMR spectroscopy confirmed successful conjugation of SMA and PAEEI-PEG. The drug loading efficiency was in the range of 34.01-72.75% and drug release lasted for 120 h. The CMC value of the micelles was affected by the SMA/PAEEI-PEG ratio and was in the range of 29.85-14.28 µg/ml. The DTX-loaded micelles showed five times more cytotoxicity than the free drug. The DTX loaded micelles were more effective in tumor growth suppression in vivo and the animals showed an enhanced rate of survival. CONCLUSION: The results show that the SMA-PAEEI-PEG micelles of DTX could potentially provide a suitable parenteral formulation with more stability, higher cytotoxicity, and improved antitumor activity.

Poly (butylene adipate-co-butylene terephthalate) nanoparticles prepared by electrospraying technique for docetaxel delivery in ovarian cancer induced mice.

OBJECTIVE: Ovarian cancer is still a major cause of morbidity and mortality. Docetaxel (DTX) is one of the most notable cytotoxic agents for treatment of ovarian cancer. However, its side effects proposed considerable problems to the patients. SIGNIFICANCE: Polymeric nanoparticles (NPs) of poly (butylene adipate-co-butylene terephthalate) (Ecoflex®), a biodegradable and biocompatible polymer, were prepared for the first time by the upgradeable electrospraying technique. METHODS: The formulation and procedure variables were optimized using Design Expert software, and effect of each variable on particle size, particle size distribution, drug entrapment efficiency, and drug release of the NPs were evaluated. Then, in vitro cytotoxicity, cellular uptake, X-ray diffraction pattern, and morphological characteristics of the optimized NPs were evaluated. Finally, in vivo efficacy of the DTX-loaded NPs was evaluated on tumor bearing nude mice. RESULTS: The optimum condition for production of NPs included voltage of 20 kV, 12 cm distance between electrodes, feeding rate of 1 mL/hr, polymer to drug ratio of 3:1, 1 w/v% of Pluronic-F127 and dichloromethane to dimethyl formamide ratio of 2.7:1. Fluorescent microscopy test showed the NPs were successfully up-taken by ovarian cancer cells. In vitro cytotoxicity test confirmed no cytotoxic effect caused by blank NPs, while cell viability of the DTX loaded NPs was significantly lower than the free DTX (p < .05). The NPs significantly enhanced anti-tumor efficacy of the drug in nude mice (p < .05). CONCLUSION: The Ecoflex® NPs could potentially provide a suitable alternative for currently available formulations of DTX.

A Rapid and Sensitive HPLC Method for Quantitation of Paclitaxel in Biological Samples using Liquid-Liquid Extraction and UV Detection: Application to Pharmacokinetics and Tissues Distribution Study of Paclitaxel Loaded Targeted Polymeric Micelles in Tumor Bearing Mice.

A simple, rapid, and sensitive reversed-phase HPLC method was developed and validated for determination of paclitaxel (PTX) in plasma, various organs and tumor tissues of tumor-bearing mice. Tissue specimens of liver, kidneys, spleen, lungs, heart and tumor were separately homogenized in normal saline. Plasma or tissue homogenate (250 µl) containing PTX and internal standard (diazepam) were extracted by diethyl ether (6 ml). The separation was achieved on a µ-Bondapak C18 HPLC column using sodium acetate buffer solution (0.01 M)/acetonitrile (58/42 v/v) at pH 5 ± 0.1 and flow rate of 1.9 mL/min. The effluent was monitored at 227 nm and column temperature was adjusted at 58ºC. The internal standard and PTX were eluted at 4.2 and 5.2 min, respectively and no interfering peaks were observed. Calibration curves were linear over the concentration range of 0.25-10 µg/ml of PTX in plasma and 0.3-20 µg/ml PTX in tissue homogenates with acceptable precision and accuracy (<15%). The mean recoveries of the drug after plasma extraction was 87.4% ± 3.6 while those of tissue homogenates ranged from 62.1± 4.5 to 75.5± 3.2 depending on the type of tissues studied. PTX was stable in samples with no evidence of degradation during 3 freeze-thaw cycles and 3 months storage at -70 °C. The developed HPLC method was applied to quantify PTX in the mouse plasma and tissues after intravenous administration of 10 mg equivalent PTX/Kg dose of PTX-loaded tocopherol succinate-chitosan-polyethylene glycol-folate (TS-CS-PEG-FA) micelles formulation or Anzatax® (Cremophor® EL- based formulation of PTX) to female Balb/c mice.

Co-delivery of paclitaxel and α-tocopherol succinate by novel chitosan-based polymeric micelles for improving micellar stability and efficacious combination therapy.

The aim of this study was to develop chitosan derivative polymeric micelles for co-delivery of paclitaxel (PTX) and α-tocopherol succinate (α-TS) to the cancer cells to improve the therapeutic efficiency and reduce side effects of PTX. In this study, amphiphilic tocopheryl succinate-grafted chitosan oligosaccharide was synthesized and physically loaded by PTX and α-TS with entrapment efficiency of 67.9% and 73.2%, respectively. Physical incorporation of α-TS into the micelles increased the hydrophobic interaction between PTX and the micelles core, which improved micelle stability, reduced the micelle size and also sustained the PTX release from the micelles. The mean particle size and zeta potential of αTS/PTX-loaded micelles were about 133 nm and +25.2 mV, respectively, and PTX release was completed during 6-9 d from the micelles. Furthermore, the cytotoxicity of α-TS/PTX-loaded micelles against human ovarian cancer cell line cancer cell in vitro was higher than that of PTX-loaded micelles and the free drug solution. Half maximal inhibitory concentration values of PTX after 48-h exposure of the cells to the PTX-loaded micelles modified and unmodified with α-TS were 110 and 188 ng/ml, respectively.

Eudragit nanoparticles loaded with silybin: a detailed study of preparation, freeze-drying condition and in vitro/in vivo evaluation.

The objective of this work was use of silybin nanoparticles in treatment of ulcerative colitis (UC). Eudragit RL PO nanoparticles loaded with silybin were produced using solvent-evaporation emulsification technique. Then, they were coated by Eudragit FS30D. Drug release was studied in different physiological environments. Colitis was induced by 4% of acetic acid in rats which received freeze-dried nanoparticles of silybin (75 mg/kg/day), dexamethasone (1 mg/kg/day), blank nanoparticles and normal saline orally for 5 days. Then macroscopic, histopathological evaluation and biochemical analysis, including myeloperoxidase (MPO) activity, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels in colon tissues were determined using enzyme-linked immunosorbent assay (ELISA) kits. Macroscopic and histopathological scores were improved by the optimised nanoparticles. The optimised nanoparticles had a particle size of 109 ± 6 nm, zeta potential of 15.4 ± 2 mV, loading efficiency of 98.3 ± 12% and release efficiency of 40.8 ± 5.5% at 24 h. TNF-α, IL-6 and MPO activity were reduced significantly by nanoparticles compared to control group (p < 0.05).

Preparation and evaluation of a controlled drug release of repaglinide through matrix pellets: in vitro and in vivo studies.

Repaglinide, an oral antidiabetic agent, has a rapid onset of action and short half-life of approximately 1 h. Designing a controlled release dosage form of the drug is required to maintain its therapeutic blood level and to eliminate its adverse effects, particularly the hypoglycaemia. Repaglinide sustained release matrix pellets consisting of Avicel, lactose and different polymers were prepared using extrusion-spheronisation method. The effect of different formulation components on in vitro drug release were evaluated using USP apparatus (paddle) for 12 h in phosphate buffer. The optimised formulation was orally administrated to normal and STZ induced diabetic rats. Most pellet formulations had acceptable physical properties with regard to size distribution, flowability and friability. Repaglinide pellets comprising Avicel 50%, lactose 47% and SLS 1% were released 94% of its drug content after 12 h. The optimised formulation was able to decrease blood glucose level in normal rats and those with diabetes throughout 8-12 h.

Effect of Tamarindus indica L. fruit pulp and seed extracts on experimental ulcerative colitis in rats.

Background and purpose: Tamarindus indica L. which has anti-inflammatory, radical scavenging, and ulcer healing effects can be useful for the alleviation of inflammatory bowel disease (IBD). Therefore, the effects of T. indica fruit pulp (TIPE) and seed extracts (TISE) were investigated on experimental colitis. Experimental approach: TIPE and TISE (125, 250, and 500 mg/kg) were made by maceration (ethanol/water: 80/30) and administered to male Wistar rats with acetic acid-induced colitis. Prednisolone (4 mg/kg) and mesalazine (100 mg/kg) were used as reference drugs. The colon tissues were examined for macroscopic and pathologic parameters and myeloperoxidase (MPO) and malondialdehyde (MDA) values. Findings/Results: The total phenols were 45.7 ± 1.1 and 453.0 ± 3.3 mg/g in terms of gallic acid for TIPE and TISE, respectively. Both of the extracts significantly improved most of the investigated parameters including body weight loss, the weight of colons, indices of ulcers, and total colitis. MPO activity and MDA in the treatment groups (except for TIPE at 125 mg/Kg) significantly decreased compared to the control. Conclusion and implications: Both TIPE and TISE were effective in the treatment of colitis however it seems that the effective ingredients were more concentrated in seeds rather than pulp extract so the highest dose of seed extract had a competitive effect with reference drugs. More studies are needed to introduce T. indica as a suitable complementary medicine or food for patients with IBD.

Platelet-cloaked alginate-poly (ß-amino ester) a novel platform bioinspired polyelectrolyte nanoparticle for targeted delivery of carboplatin in breast cancer: An in vitro/in vivo study.

Triple-negative breast cancer (TNBC) has poor prognosis. Carboplatin (Crb) is a widely used chemotherapeutic agent, in TNBC but with serious systemic toxicity and poor tumor targeting. Bioinspired drug-loaded platelets (Plt) and Plt-coated nanocarriers evade macrophage phagocytosis by membrane proteins like CD47. The goal of this study was preparation of a novel alginate-poly (ß-amino ester) (PßAE) nanoparticles (NPs) for targeted delivery of Crb to TNBC cells by developing and comparison of two bioinspired carriers of Plt membrane (PltM) coated Crb-loaded alginate-poly (ß-amino ester) nanoparticles (PltM@Crb-PßAE-ALG NPs) and Plt loaded Crb (Plt@Crb). The NPs were prepared by ionic gelation and subsequently were coated by platelet membrane using ultra-sonication method. The loading efficiency, release profile, and in vitro cytotoxicity of both formulations were evaluated on HUVEC and 4 T1 cells. Additionally, the in vivo tumor targeting, therapeutic efficacy, and organ toxicity of the two formulations were assessed in a murine tumor model. Results showed both Plt@Crb and (PltM@Crb-PßAE-ALG NPs) exhibited high drug loading efficiency, sustained release, enhanced cytotoxicity against 4 T1 cells, and decreased cytotoxicity in normal cells (HUVEC) in vitro. In vivo studies revealed that although both formulations considerably improved tumor inhibition compared to free Crb, but the PltM@Crb-PßAE-ALG NPs demonstrated superior cytotoxicity and therapeutic efficacy, thanks to improved Crb's internalization efficiency, enhanced stability, and controlled release properties.

Magnetophoretic Intranasal Drug-loaded Magnetic Nano-aggregates as a Platform for Drug Delivery in Status Epilepticus.

BACKGROUND: Status epilepticus is associated with substantial morbidity and neuronal necrosis, and the duration of the seizure would affect its following complications. Eliminating the duration would have valuable outcomes; however, the presence of BBB is an obstacle. The purpose of the current study was to achieve a nose-to-brain magnetic drug delivery system to accelerate the onset of action, and to reduce the mucociliary clearance via implementing the magnetic field. MATERIALS AND METHODS: The drug-entrapped magnetic nanoaggregates were prepared via a 2-step method, synthesis of the magnetic nanoparticles and drug loading. Optimization of the variables, including ammonium hydroxide:water ratio, beta-cyclodextrin%, duration of the mixing time, amount of Pluronic, and drug:magnetic nanoaggregates mass ratio was performed according to particle size, PDI, zeta potential, release profile and entrapment efficiency. The efficacy of optimized formulation was assessed in the animal model. RESULTS: According to the analysis performed by the software, drug-to-nanoparticle ratio and the duration of mixing time were found to be significantly effective (p < 0.05) for entrapment efficiency and particle size distribution, respectively. The optimum formulation with an approximate average size of 581 nm and 61% entrapment efficiency was obtained, which released about 80% of its drug content within the first 20 minutes. The in vivo efficacy was significantly improved (p < 0.05) by administration of magnetic nanoaggregates in the presence of a simple external magnet placed on the glabellar region of the animals, compared to the control groups. CONCLUSION: This drug delivery system could be suggested as a fast-acting alternative for seizure cessation in status epilepticus emergencies.

Ameliorative Effect of Aqueous and Hydroalcoholic Extracts of Scrophularia striata Boiss. on Murine Model of Experimental Colitis.

Background: Scrophularia striata Boiss. (S. striata) is a flowering plant with several therapeutic properties including antiinflammatory, antioxidant, antimicrobial, and wound-healing activity. Regarding the side effects of drugs conventionally used for inflammatory bowel disease (IBD) treatment, we investigated the anticolitis properties of aqueous (SSAE) and hydroalcoholic (SSHE) extracts of S. striata on experimental colitis. Materials and Methods: The colitis was induced using acetic acid (3%) and 2 h before ulcer induction, each group of rats received orally three doses (150, 300, and 600 mg/kg, p.o.) of SSAE or SSHE for the next 5 days. Dexamethasone (1 mg/kg, i.p.) and mesalazine (100 mg/kg, p.o.) were used as reference drugs. Different parameters including weight of colon/height, ulcer index, total colitis index, levels of myeloperoxidase (MPO) and malondialdehyde (MDA) were investigated. Results: Total phenolic contents were 4.3 ± 0.2 and 7.1 ± 0.4 mg/g equivalent to gallic acid for SSAE and SSHE respectively. Three applied doses of SSHE and the highest dose of SSAE (600 mg/kg) could reduce all the macroscopic and pathologic indices of colitis and the levels of MPO and MDA. Two lesser doses of SSAE (150, 300 mg/kg) however, couldn't diminish the histopathologic features of colitis and the values of MPO and MDA. Conclusions: S. striata, especially SSHE, which also contained more phenolic compounds, had an ameliorating effect on ulcerative colitis and possibly exerts this effect through its antioxidant, antiinflammatory and wound healing properties. Further investigations are required to introduce this plant as a novel alternative herbal drug for colitis treatment.

Preparing simvastatin nanoparticles by a combination of pH-sensitive and timed-release approaches for the potential treatment of ulcerative colitis.

In this study, an emulsion solvent evaporation method was used to produce Eudragit RL (ERL) nanoparticles (NPs) loaded with simvastatin (SIM) for the treatment of ulcerative colitis (UC). Accordingly, the effects of different formulation variables on the properties of NPs were evaluated using the Box-Behnken design. The optimized NPs were then coated by Eudragit FS30D (EFS30D). Drug release was studied in different physiological environments. Colitis was induced by 3% of acetic acid in rats, which received NPs of SIM (10 mg/kg/day), mesalazine (150 mg/kg/day), blank NPs and normal saline orally for 5 days. Macroscopic histopathological evaluation and biochemical analysis, including myeloperoxidase (MPO) activity and malondialdehyde (MDA) level in the colon tissues, were carried out in this study. The optimized SIM-ERL NPs showed the particle size of 182.48 ± 4.57 nm, the polydispersity index of 0.29 ± 0.12, the zeta potential of 26.45 ± 4.57 mV, drug loading % of 34.64 ± 0.48, the encapsulation efficiency % of 98.68 ± 0.69, and the release efficiency % of 35.78 ± 1.37. Coating the optimized NPs with EFS30D caused an increase in particle size and a decrease in the zeta potential of NPs. The optimized SIM-EFS30D/RL NPs improved the macroscopic and histopathological scores. Also, MPO activity and MDA level were reduced significantly by NPs, as compared to the control group. Therefore, this drug delivery system can be an alternative to the previous treatments of UC.

Salvia officinalis leaf extracts protect against acute colitis in rats.

Background and purpose: Inflammatory bowel disease (IBD) is a chronic and multifactorial disease with unknown etiology and a decisive cure. Salvia officinalis (sage) which has anti-inflammatory, anti-oxidative, and ulcer healing properties can be useful for the treatment of IBD. Therefore, the effect of S. officinalis ethanolic extract (SOEE) and methanolic partition (SOMP) was investigated on acetic acid-induced ulcerative colitis. Experimental approach: Male Wistar rats (180-220 g) were used. SOEE (30, 60, and 120 mg/kg) and SOMP (50, 100, and 150 mg/kg) were prepared through maceration method. Prepared extracts, dexamethasone (1 mg/kg, i.p.), and mesalamine (100 mg/kg) as reference drugs and normal saline as control were administered by gavage, 2 h before colitis induction and preserved for four further days to animals. The colon tissues were examined for macroscopic and pathologic parameters and myeloperoxidase (MPO) and malondialdehyde (MDA) levels. Findings/Results: SOEE (60 and 120 mg/kg) and SOMP at all doses alleviated colitis severity and indices both in macroscopic and microscopic views. MDA and MPO activities were also significantly declined in the extracts-treated groups compared to the controls. The lowest dose of SOEE couldn't meaningfully reduce any of the parameters compared to the control group. Conclusion and implications: Both extracts of S. officinalis exerted anti-colitis effects in rats, though methanolic partition was more effective, especially at the highest dose. It seems S. officinalis could exert protection against oxidative stress and inflammatory mediators in colitis tissue. More experimental and clinical studies are required to explore the exact mechanisms and active ingredients which are involved.

Development and validation of a new robust RP-HPLC method for simultaneous quantitation of insulin and pramlintide in non-invasive and smart glucose-responsive microparticles.

Background and purpose: Since insulin and pramlintide cooperate in glucose hemostasis, co-administration and quantitation of them in pharmaceutical preparations are imperative. A simple, rapid, sensitive, and isocratic RP-HPLC method was developed and validated for simultaneous quantitation of insulin and pramlintide in loading and in-vitro release studies of a glucose-responsive system to improve the control of hyperglycemic episodes in diabetic patients. Experimental approach: The isocratic RP-HPLC separation was achieved on a C18 µ-Bondopak column (250 mm × 4.6 mm) using a mobile phase of water:acetonitrile:trifluoroacetic acid (65:35:0.1%) at a flow rate of 1 mL/min in an ambient temperature. Both proteins were detected using a UV detector at 214 nm. The method was validated for specificity, linearity, precision, accuracy, the limit of detection, the limit of quantification, and robustness. Findings/Results: Linearity was obtained in the concentration range of 30 to 360 µg/mL for insulin and 1.5 to 12 µg/mL for pramlintide. The results were validated statistically and recovery studies confirmed the great accuracy and precision of the proposed method. The robustness of the method was also confirmed through small changes in pH, mobile phase composition, and flow rate. Conclusion and implications: The method was found to be simple, specific, precise, and reproducible. It was applied for the determination of loading capacity, entrapment efficiency, and in-vitro release studies of insulin and pramlintide in a smart glucose-responsive microparticle. Co-delivery of insulin and pramlintide could be a new intervention in diabetes management and concurrent quantitation of these two proteins is, therefore, essential.