فهرست مطالب

  • Volume:10 Issue: 2, 2020
  • تاریخ انتشار: 1399/01/20
  • تعداد عناوین: 20
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  • Iti Chauhan*, Mohd Yasir, Madhu Verma, Alok Pratap Singh Pages 150-165

    Nanostructured lipid carriers (NLCs) are novel pharmaceutical formulations which arecomposed of physiological,biocompatible lipids,surfactants,co-surfactants. Over time,as a second generation lipid nanocarrier NLC has emerged as an alternative to first generationnanoparticles. This review article highlights the structure,composition,various formulationmethodologies,and characterization of NLCs which are prerequisites in formulating a stabledrug delivery system. NLCs hold an eminent potential in pharmaceuticals,cosmetics marketbecause of extensive beneficial effects like skin hydration,occlusion,enhanced bioavailability,and skin targeting. This article aims to evoke an interest in the current state of art NLC bydiscussing their promising assistance in topical drug delivery system. The key attributes of NLCthat make them a promising drug delivery system are ease of preparation,biocompatibility,thefeasibility of scale up,non-toxicity,improved drug loading,and stability.

    Keywords: Nanostructure lipid carrier, Lipid, Topical, Skin
  • Dipak Dilip Gadade*, Sanjay Sudhakar Pekamwar Pages 166-183

    Colloidal nanoparticulate technology has been described in the literature as a versatile drug delivery system. But it possesses some inherent lacunae in their formulation. Cyclodextrins (CDs) have been extensively reported for the solubility enhancement of poorly water-soluble drugs. The CDs can cause intervention in aspects related to nanoparticles (NPs) that include improving drug loading in nano-system, improving stability, site-specific/targeted drug delivery, improving solubility profile and absorption of the drug in nanosystem with consequent improvement in bioavailability, with the possibility of controlled release, safety and efficacy. They find application in for simultaneous diagnosis and therapeutics for better treatment procedures. The current communication is focused on the application of CDs to overcome troubles in nanoparticulate formulation and enhancement of their performance. It also envisages the theranostic aspects of CDs.

    Keywords: Cyclodextrinm, Nanoparticles, Solubility, Theranostics, Stability, Controlled release
  • Velid Unsal*, Tahir Dalkiran, Mustafa Çiçek, Engin Kölükçü Pages 184-202

    Cadmium (Cd) is a significant ecotoxic heavy metal that adversely affects all biological processes of humans, animals and plants. Exposure to acute and chronic Cd damages many organs in humans and animals (e.g. lung, liver, brain, kidney, and testes). In humans, the Cd concentration at birth is zero, but because the biological half-life is long (about 30 years in humans), the concentration increases with age. The industrial developments of the last century have significantly increased the use of this metal. Especially in developing countries, this consumption is higher. Oxidative stress is the imbalance between antioxidants and oxidants. Cd increases reactive oxygen species (ROS) production and causes oxidative stress. Excess cellular levels of ROS cause damage to proteins, nucleic acids, lipids, membranes and organelles. This damage has been associated with various diseases. These include cancer, hypertension, ischemia/perfusion, cardiovascular diseases, chronic obstructive pulmonary disease, diabetes, insulin resistance, acute respiratory distress syndrome, idiopathic pulmonary fibrosis, asthma, skin diseases, chronic kidney disease, eye diseases, neurodegenerative diseases (amyotrophic lateral sclerosis, Parkinson’s disease, Alzheimer’s disease, and Huntington disease). Natural antioxidants are popular drugs that are used by the majority of people and have few side effects. Natural antioxidants play an important role in reducing free radicals caused by Cd toxicity. Our goal in this review is to establish the relationship between Cd and oxidative stress and to discuss the role of natural antioxidants in reducing Cd toxicity.

    Keywords: Cadmium, Oxidative stress, ROS, Natural antioxidants
  • Abdul Raheem Thayyil*, Thimmasetty Juturu, Shashank Nayak, Shwetha Kamath Pages 203-212

    Pharmaceutical co-crystals are novel class of pharmaceutical substances, which possess an apparent probability of advancement of polished physical properties offering stable and patentable solid forms. These multi-component crystalline forms influence pertinent physicochemical parameters like solubility, dissolution rate, chemical stability, physical stability, etc. which in turn result in the materials with superior properties to those of the free drug. Co-crystallization is a process by which the molecular interactions can be altered to optimize the drug properties. Co-crystals comprise a multicomponent system of active pharmaceutical ingredient (API) with a stoichiometric amount of a pharmaceutically acceptable coformer incorporated in the crystal lattice. By manufacturing pharmaceutical co-crystals, the physicochemical properties of a drug can be improved thus multicomponent crystalline materials have received renewed interest in the current scenario due to the easy administration in the pharmaceutical industry. There is an immense amount of literature available on co-crystals. However, there is a lack of an exhaustive review on a selection of coformers and regulations on co-crystals. The review has made an attempt to bridge this gap. The review also describes the methods used to prepare co-crystals with their characterization. Brief description on the pharmaceutical applications of co-crystals has also been incorporated here. Efforts are made to include reported works on co-crystals, which further help to understand the concept of co-crystals in depth.

    Keywords: Co-crystals, Evaluation of co-crystals, Hansen solubility parameter, Liquid assisted grinding, Preparation of co-crystals, Solvent evaporationtechnique
  • Mahmoud Abudayyak*, Elif Guzel, Gül Özhan Pages 213-220
    Purpose

    The wide application of cupric oxide nanoparticles (copper (II) oxide, CuO-NPs) in various fields has increased exposure to the kind of active nanomaterials, which can cause negative effects on human and environment health. Although CuO-NPs were reported to be harmful to human, there is still a lack information related to their toxic potentials. In the present study, the toxic potentials of CuO-NPs were evaluated in the liver (HepG2 hepatocarcinoma) and intestine (Caco-2 colorectal adenocarcinoma) cells.

    Methods

    After the characterization of particles, cellular uptake and morphological changes were determined. The potential of cytotoxic, genotoxic, oxidative and apoptotic damage was investigated with several in vitro assays.

    Results

    The average size of the nanoparticles was 34.9 nm, about 2%-5% of the exposure dose was detected in the cells and mainly accumulated in different organelles, causing oxidative stress, cell damages, and death. The IC50 values were 10.90 and 10.04 µg/mL by MTT assay, and 12.19 and 12.06 µg/mL by neutral red uptake (NRU) assay, in HepG2 and Caco-2 cells respectively. Apoptosis assumes to the main cell death pathway; the apoptosis percentages were 52.9% in HepG2 and 45.5% in Caco-2 cells. Comet assay result shows that the highest exposure concentration (20 µg/mL) causes tail intensities about 9.6 and 41.8%, in HepG2 and Caco-2 cells, respectively.

    Conclusion

    CuO-NPs were found to cause significant cytotoxicity, genotoxicity, and oxidative and apoptotic effects in both cell lines. Indeed, CuO-NPs could be dangerous to human health even if their toxic mechanisms should be elucidated with further studies.

    Keywords: Copper oxide, Nanoparticle, DNA damage, Cytotoxicity, Oxidative stress, Apoptosis
  • Nadia Karimi, Kamaran Mansouri, Mohammad Soleiman Beigi, Ali Fattahi* Pages 221-232
    Purpose

    Developing chemotherapy with nanoplatforms offers a promising strategy for effective cancer treatment. In the present study, we propose a novel all-trans retinoic acid (ATRA) grafted poly beta-amino ester (PBAE) copolymer for preparing nanoparticles (NPs).

    Methods

    ATRA grafted PBAE (ATRA-g-PBAE) copolymer was synthesized by grafting ATRA to PBAE; it was characterized by proton nuclear magnetic resonance, Fourier transform infrared, and thermogravimetric analysis. ATRA-g-PBAE NPs were prepared by the solvent displacement method. Design-Expert software was employed to optimize size of NPs. The morphology was evaluated by transmission electron microscope, and ultraviolet-visible spectroscopy was applied for drug release. Cytotoxicity was evaluated toward HUVEC cell line, and the 3D collagencytodex model was used to evaluate anti-angiogenic property of PBAE, ATRA, and NPs.

    Results

    The optimum size of the NPs was 139.4 ± 1.41 nm. After 21 days, 66.09% ± 1.39 and 42.14% ± 1.07 of ATRA were released from NPs at pH 5.8 and 7.4, respectively. Cell culture studies demonstrated antiangiogenic effects of ATRA-g-PBAE NPs. Anti-angiogenesis IC50 was 0.007 mg/mL for NPs (equal to 0.002 mg/mL of ATRA) and 0.005 mg/mL for free ATRA.

    Conclusion

    This study proposes the ATRA-g-PBAE NPs with inherent anti-angiogenic effects as promising carrier for anticancer drugs with purpose of dual drug delivery.

    Keywords: All-trans retinoic acid, Anti-angiogenesis, Nanoparticles, Poly (β-amino ester), Response surfacemethodology
  • Samira Nekoufar, Ahmad Fazeli, MohammadReza Fazeli* Pages 233-238
    Purposes

    Solubilization of inclusion bodies expressed in E. coli is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high purity and yield. The aim of this study was to examine the benefit of organic solvents such as alcohols in solubilization of recombinant interferon β-1b inclusion bodies.

    Methods

    Effect of important parameters inclusion pH, concentration and type of denaturant and concentration of alcoholic solvents were optimized to formulate a suitable solubilization buffer and investigate their effect on solubilization of interferon β-1b inclusion bodies.

    Results

    Our findings showed the acidic pH in the range of 2-3 is more suitable than alkaline pH >12 for solubilization and achieving higher content of interferon β-1beta and pure recombinant protein. We have also demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon β-1b at pH of 2-3. The interferon concentration was 2.35 mg per 100 mg IB when we used 40% (v/v) 1-propanol and 20% (v/v) 2-butanol into the buffer solution as well.

    Conclusion

    The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area.

    Keywords: Interferon beta-1b, Inclusion body, Solubilization, Organic solvent, Alcohol
  • Anayatollah Salimi, Behzad Sharif Makhmal Zadeh*, Salar Godazgari, Abbas Rahdar Pages 239-246
    Purpose

    Azelaic acid is a natural keratolytic, comedolytic, and antibacterial drug that is used to treat acne. The topical application of azelaic acid is associated with problems such as irritation and low permeability. For dissolving, the problem is that microemulsion (ME) is used as a drug carrier. The aim of this study was to increase the azelaic acid affinity in the follicular pathway through ME.

    Methods

    Azelaic acid-loaded MEs were prepared by the water titration method. The properties of the MEs included formulation stability, particle size, drug release profile, thermal behavior of MEs, the diffusion coefficient of the MEs and skin permeability in the non-hairy ear skin and hairy abdominal skin of guinea pig were studied in situ.

    Results

    The MEs demonstrated a mean droplet size between 5 to 150 nm. In the higher ratios of surfactant/co-surfactant, a more extensive ME zone was found. All MEs increased the azelaic acid flux through both hairy and non-hairy skin compared with an aqueous solution of azelaic acid as a control. This effect of the ME was mainly dependent on the droplet diffusion coefficient and hydrodynamic radius. MEs with a higher diffusion coefficient demonstrated higher azelaic acid flux through hairy and non-hairy skin. Drug flux through both skins was affected by the surfactant/co-surfactant ratio in that the higher ratio increased the azelaic acid affinity into the follicular pathway.

    Conclusion

    Finally, the ME with the highest droplet diffusion coefficient and the lowest surfactant/co-surfactant ratio was the best ME for azelaic acid delivery into the follicular pathway

    Keywords: Azelaic acid, Microemulsion, Phase diagram, Follicular drug delivery, Acne vulgaris, Acne treatment
  • Madikeri Manjunath Charithra, Jamballi Gangadharappa Gowda Manjunatha*, Chenthattil Raril Pages 247-253
    Purpose

    Estriol (ERL) is a type of hormone among the groups of estrogen hormone that was detected through the voltammetric technique by constructing an electrochemical sensor based on the octoxynol-9 modified graphite paste electrode (OXL-9MGPE).

    Methods

    Using the strategy of cyclic voltammetry (CV) and differential pulse voltammetry (DPV) with a bare graphite paste electrode (BGPE) immobilized with OXL-9, ERL electro-oxidation has been assessed in 0.2 M phosphate buffer solution (PBS) of pH 6.0. The fabricated electrode has substantial electrochemical sensing efficiency, and the ERL oxidation at the OXL-9MGPE was the irreversible process. The surface morphological characteristics of BGPE and OXL-9MGPE were differentiated with the help of field emission scanning electron microscopy (FE-SEM).

    Results

    The impact of various factors such as scan rate, pH, reproducibility, repeatability, and stability on the electro-oxidation of ERL was evaluated. Techniques of CV and DPV were utilized to determine ERL, dopamine (DAN), and uric acid (URA) simultaneously with the projected sensor. The peak current was varied with ERL concentration in the range from 4×10-5 to 1.2×10-4 M at OXL-9MGPE. From this, the detection limit 1.4×10-6 M and limit of quantification (LOQ) 4.7×10-6 M have been attained.

    Conclusion

    As a result, OXL-9MGPE was successfully achieved as an electrochemical detector for the electro analysis of ERL via the CV technique.

    Keywords: Electrochemical sensor, Estriol, Graphite paste electrode, Octoxynol-9 immobilization
  • Rasool Mirzaei Seveiri, Masoud Hamidi, Cédric Delattre, Hamid Sedighian, Guillaume Pierre, Babak Rahmani, Sina Darzi, Clément Brasselet, Fatemeh Karimitabar, AliRaza ghpoor, Jafar Amani* Pages 254-263
    Purpose

    Due to the potential industrial and therapeutic applications of the yeast exopolysaccharides (EPSs), there has been an increasing demand to assess these biopolymers with improved characteristics. This study aimed to characterize the EPSs from Rhodosporidium babjevae (ATCC 90942 and IBRC-M 30088) as well as to evaluate their possible antioxidant, emulsifying and antiproliferative activities.

    Methods

    Rhodosporidium babjevae was cultured for 5 days and following isolation of supernatant, EPSs precipitated with adding of cold absolute ethanol and freeze-dried. The EPSs chemical structure was determined by FT-IR, SEM, HPLC-SEC and GC-MS. Additionally the solubility, water holding capacity and emulsifying activity of EPSs were evaluated. In vitro, antioxidant activity was investigated against DPPH, superoxide and hydroxyl radicals. Finally the EPSs consequence on the cell proliferation of human breast adenocarcinoma (MCF-7) and Madin-Darby canine kidney (MDCK) cell lines was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test.

    Results

    R. babjevae excreted 1.6±0.2 g/L of the EPSs. The EPSs had three fractions with molecular weights of 1.02 ×106 , 5×105 and 2×105 Da. Mannose and glucose were found as the main monosaccharides of the EPSs (84:16 mol%, respectively). The EPSs exhibited emulsifying activity on sun flower oil. The scavenging activities were found to be dose-dependent and higher than hyaluronic acid. Significant difference among the EPSs treatments on the proliferation of MCF-7 and MDCK cell lines was not observed (P>0.05).

    Conclusion

    These results show the interesting potential of the EPSs from R. babjevae as biocompatible compounds for using in food and pharmaceutical fields.

    Keywords: Exopolysaccharides, Rhodosporidium babjevae, Antioxidant activity, Emulsifying activity, Antiproliferative activity
  • Nafiseh Paydarnia, Behzad Mansoori, Davoud Esmaeili, Tohid Kazemi, Mahyar Aghapour, Khalil Hajiasgharzadeh, Nazila Alizadeh, Behzad Baradaran Pages 264-270
    Purpose

    Helicobacter pylori is recognized as one of the prevalent causes of human gastric infection. In the present study, the role of mixed immunization with H. pylori lipopolysaccharide (LPS) and recombinant cytotoxin-associated gene A (rCagA) as a stimulator of host immune responses was determined.

    Methods

    BALB/c mice were immunized with different formulations by the systemic administration at 14-day intervals. The effects of the formulations plus CpG adjuvants were assessed before and post-immunization in separated studies. Moreover, the expression of Th1/Th2 cytokines was quantified in sera of immunized mice using reverse transcription polymerase chain reaction (RTPCR) test and the protein levels confirmed with enzyme linked immunosorbent assay (ELISA). Finally, the specific antibody levels in sera were studied by ELISA and the tendency of cellular response was examined by IgG1/IgG2a ratio.

    Results

    Data of Western blotting verified the presence of constructed protein. Analysis of lymphocyte proliferation showed that CpG-conjugated rCagA increases lymphocytes proliferation compared to the control group. Also, it was shown that formulations containing LPS and rCagA promote a Th1 response indicated by interferon-gamma expression and induced Th1/ Th2 balance. Additionally, the specific IgG1, total IgG and IgG2a levels elevated in response to all treatments. Ultimately, the IgG2a/IgG1 ratio in the mice immunized with rCagA-containing formulations increased.

    Conclusion

    These results indicated that rCagA protein carried with CpG adjuvant not only maintained its antigenicity throughout the experiment but also induced robust Th1-biased immune responses. Therefore, it holds promise for the production of an efficient vaccine against H. pylori infection.

    Keywords: Helicobacter pylori, Th1 immune response, Immunization, Lipopolysaccharide, Recombinant cytotoxinassociated gene A
  • Leila Dinparast, Siavoush Dastmalchi* Pages 271-277
    Purpose

    Despite the discovery and synthesis of several anticancer drugs, cancer is still a major life threatening incident for human beings after cardiovascular diseases. Toxicity, severe side effects, and drug resistance are serious problems of available commercial anticancer drugs. Coumarins are synthetic and natural heterocycles that show promising antiproliferative activities against various tumors. The aim of this research is to computationally study the coumarin derivatives in order to develop reliable quantitative structure-activity relationship (QSAR) models for predicting their anticancer activities.

    Methods

    A data set of thirty one coumarin analogs with significant antiproliferative activities toward HepG2 cells were selected from the literature. The molecular descriptors for these compounds were calculated using Dragon, HyperChem, and ACD/Labs programs. Genetic algorithm (GA) accompanied by multiple linear regression (MLR) for simultaneous feature selection and model development was employed for generating the QSAR models.

    Results

    Based on the obtained results, the developed linear QSAR models with three and four descriptors showed good predictive power with r2 values of 0.670 and 0.692, respectively. Moreover, the calculated validation parameters for the models confirmed the reliability of the QSAR models.

    Conclusion

    The findings of the current study could be useful for the design and synthesis of novel anticancer drugs based on coumarin structure.

    Keywords: Coumarin, Cancer, Antiproliferative, QSAR, GA-MLR
  • Yasamin Pahlavan, Naser Samadi, Khalil Ansarin, Alireza Khabbazi* Pages 278-283
    Purpose

    Survivin is critical for proliferation, maturation, homeostasis and differentiation of effector and memory lymphocytes. In this study the baculoviral inhibitors of apoptosis proteins (IAPs) repeat containing 5 (BIRC5) mRNA, survivin, and phosphorylated survivin expression were evaluated in peripheral blood mononuclear cells (PBMCs), and plasma of patients with Behcet’s disease (BD).

    Methods

    In this study, 26 Iranian Azari patients diagnosed with BD and 30 healthy controls were recruited. Total RNA was extracted from PBMCs. The expression level of survivin was measured by quantitative real-time polymerase chain reaction (PCR). Survivin plasma levels were measured using survivin Enzyme-linked immunosorbent assays. Also, western blotting analysis was performed to measure phosphorylated-survivin and survivin levels in PBMCs and plasma of patients with BD.

    Results

    In a pilot study, we showed that BIRC5 gene expression increased in BD patients compared with healthy controls (P<0.05). Western blotting analysis indicated that there was an increase in phosphorylated survivin expression in PBMCs of BD patients. Our data from western blot analysis showed survivin level in plasma samples of BD patients was similar to healthy controls. No significant differences were observed between plasma survivin levels in the BD patients compared with control group (P>0.05). The expression of phosphorylated survivin at Thr34 in PBMCs of BD patients with active disease was increased. Plasma phosphorylated survivin levels in having BD patients were also downregulated compared to healthy individuals.

    Conclusion

    Analysis of PBMCs indicated increasing expression level of phosphorylated survivin in PBMCs of BD patients. There was also a downregulation in phosphorylated survivin levels in plasma of BD patients.

    Keywords: Autoimmunity, Apoptosis, Lymphocytes, Inhibition of apoptosisproteins (IAPs), Behcet’s disease
  • Sheyda Shaafi, Soraiya Ebrahimpour Koujan, Mohammad Khalili*, SeyadMorteza Shamshirgaran, Mazyar Hashemilar, Aliakbar Taheraghdam, SeyedKazem Shakouri, Elyar Sadeghi Hokmabadi, Yaeghoub Ahmadi, Mehdi Farhoudi, Nasim Rezaeimanesh, Daryoush Savadi Osgouei Pages 284-289
    Purpose

    Stroke is one of the most common conditions causing death. There have been few studies examining the effects of alpha lipoic acid (ALA) on stroke patients. In this regard, the present randomized controlled clinical trial was conducted to examine the effects of ALA supplementation on serum albumin, and inflammatory and oxidative stress markers in stroke patients.

    Methods

    The present paralleled randomized controlled clinical trial involved 42 stroke patients who were over 40 years and under enteral feeding. The participants were randomly assigned into two groups and finally 40 patients completed the study. Patients in alpha lipoic acid group (n=19) took 1200 mg ALA supplement daily along with their meal, and participants in control group (n=21) underwent the routine hospital diet for 3 weeks. Fasting blood samples were obtained and albumin, oxidative stress, and inflammatory indices were assessed at baseline, as well as at the end of the trial.

    Results

    After 3 weeks, treatment of patients with ALA led to a significant decrease in tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) levels (P=0.01) compared to baseline. But serum levels of albumin, total antioxidant capacity (TAC), malondialdehyde (MDA), highsensitivity C-reactive protein (hs-CRP), IL-6 and TNF-α did not change significantly vs. control group (P>0.05).

    Conclusion

    ALA did not significantly change the serum levels of albumin and inflammatory as well as antioxidant capacity indices in stroke patients compared with the control group. More clinical trials with large sample sizes and long duration are needed to clarify the effects of ALA on these patients.

    Keywords: Alpha lipoic acid, Inflammation, Oxidative stress, Stroke
  • Fariba Hajifathaliha, Arash Mahboubi*, Elham Mohit, Noushin Bolourchian, Vahid Khalaj, Leila Nematollahi* Pages 290-296
    Purpose

    Poly l-lysine (PLL) has been introduced as a strengthening covering layer for alginate microcapsules which are the most convenient way for cell encapsulation. Some disadvantages of PLL such as high price and low biocompatibility have prompted scientists to find better alternatives. Linear poly ethylene imine (LPEI), thanks to its highly similar structure to PLL, could be considered as a proper cost-effective alternative. In this study LPEI and PLL were compared as covering layers of cell-loaded alginate-LPEI-alginate (cALA) and alginate-PLL-alginate (cAPA) microcapsules.

    Methods

    In addition to the physico-mechanical properties, the encapsulation efficiency, cell survival post encapsulation, cell viability, and cellular metabolic activity within the microcapsules were evaluated using trypan blue, live/dead cell staining, and MTT test, respectively.

    Results

    Physico-mechanical evaluation of the microcapsules revealed that the cell microencapsulation process did not affect their shape, size, and mechanical stability. Although the encapsulation efficiency for cALA and cAPA was not different (P>0.05), cell survival post encapsulation was higher in cALA than in cAPA (P<0.05) which could be the reason for the higher cell viability and also cellular metabolic activity within these microcapsules in comparison to cAPA.

    Conclusion

    Here, based on these results, ALA could be introduced as a preferable alternative to APA for cell encapsulation.

    Keywords: Alginic acid, Cell microencapsulation, CHOK1, Poly ethylene imine, Poly l-lysine
  • Kamal Abdolmohammadi, Tayebeh Mahmoudi, Shahrzad Nojehdehi, Lobat Tayebi, SeyedMahmoud Hashemi, Farshid Noorbakhsh, Alireza Abdollahi, Masoud Soleimani, Behrouz Nikbin, MohammadHossein Nicknam* Pages 297-306
    Purpose

    Acute pancreatitis (AP) is an inflammatory disorder distinguished by tissue injury and inflammation of the pancreas. Using paracrine potential of mesenchymal stem cells (MSCs) provides a useful clinical approach in treating inflammatory diseases. We investigated the therapeutic effects of adipose-derived MSC conditioned medium (CM) and hypoxia preconditioned adipose-derived MSC conditioned medium (HCM) in cerulein-induced AP in mice.

    Methods

    AP was induced in C57BL/6 mice by intraperitoneal injection of cerulein (75 μg/ kg/h × 7 times). One hour following the last injection of cerulein, mice were treated with intraperitoneal injection of CM and HCM (500 µL/mice/30 min × 3 times). Twelve hours following the treatment, serum levels of amylase and lipase were measured. In addition, pancreas pathological changes, immunohistochemical examinations for evaluation of IL-6 expression and pancreatic myeloperoxidase (MPO) enzyme activity were analyzed.

    Results

    The in vitro results of the morphological, differentiation and immunophenotyping analyses confirmed that hypoxia preconditioned MSCs (HP-MSCs) conserve MSCs characteristics after preconditioning. However, HP-MSCs significantly expressed high mRNA level of hypoxia inducible factor 1-α and higher level of total protein. The in vivo findings of the current study showed that CM and HCM significantly reduced the amylase & lipase activity, the severity of pancreas tissue injury and the expression of IL-6 and MPO enzyme activity compared with the AP group. However, no significant difference between CM and HCM groups was demonstrated.

    Conclusion

    Use of CM and HCM can attenuate cerulein-induced AP and decrease inflammation in the pancreas tissue in AP mice.

    Keywords: Inflammation Acute pancreatitis, Cerulein, Mesenchymal stem cell, Conditioned medium, Preconditioning
  • Ezzatollah Fathi, Behnaz Valipour, Zohreh Sanaat, Hojjatollah Nozad Charoudeh, Raheleh Farahzadi* Pages 307-314
    Purpose

    The effect of mesenchymal stem cells (MSCs) on the immortality features of malignant cells, such as hematologic cancerous cells, are controversial, and the associated mechanisms are yet to be well understood. The aim of the present study was to investigate the in vitro effect of bone marrow-derived MSCs (BMSCs) on the chronic myeloid leukemia cell line K562 through telomere length measurements, telomerase activity assessments, and hTERT gene expression. The possible signaling pathways involved in this process, including Wnt-5a/β-catenin and P53, were also evaluated.

    Methods

    Two cell populations (BMSCs and K562 cell line) were co-cultured on transwell plates for 7 days. Next, K562 cells were collected and subjected to quantitative real-time PCR, PCR-ELISA TRAP assay, and the ELISA sandwich technique for telomere length, hTERT gene expression, telomerase activity assay, and cytokine measurement, respectively. Also, the involvement of the mentioned signaling pathways in this process was reported by real-time PCR and Western blotting through gene and protein expression, respectively.

    Results

    The results showed that BMSCs caused significant decreases in telomere length, telomerase activity, and the mRNA level of hTERT as a regulator of telomerase activity. The significant presence of interleukin (IL)-6, IL-8, and transforming growth factor beta (TGF-β) was obvious in the co-cultured media. Also, BMSCs significantly decreased and increased the gene and protein expression of β-catenin and P53, respectively.

    Conclusion

    It was concluded that the mentioned effects of IL-6, IL-8, and TGF-β cytokines secreted from MSCs on K562 cells as therapeutic agents were applied by Wnt-5a/β-catenin and P53 pathways.

    Keywords: Mesenchymal stem cells, Telomere length, Telomerase activity, Cytokines, Wnt5a, β-catenin, P53 signaling pathways
  • Sara Aqmasheh, Karim Shamsasenjan*, Elham Khalaf Adeli, Aliakbar Movassaghpourakbari, Parvin Akbarzadehlaleh, Davod Pashoutan Sarvar, Hamzeh Timari Pages 315-322
    Purpose

    Mesenchymal stem cells (MSCs) release hematopoietic cytokines, growth factors, and Microvesicles (MVs) supporting the hematopoietic stem cells (HSCs). MVs released from various cells, playing a crucial role in biological functions of their parental cells. MSC-derived MVs contain microRNAs and proteins with key roles in the regulation of hematopoiesis. Umbilical cord blood (UCB) is a source for transplantation but the long-term recovery of platelets is a main problem. Therefore, we intend to show that MSC-MVs are able to improve the differentiation of UCB-derived CD34+ cells to megakaryocyte lineage.

    Methods

    In this descriptive study, MSCs were cultured in DMEM to collect the culture supernatant, which was ultracentrifuged for the isolation of MVs. HSCs were isolated from UCB using MACS method and cultured in IMDM supplemented with cytokines and MVs in three different conditions. Megakaryocyte differentiation was evaluated through the expression of specific markers and genes after 72 hours, and the data was analyzed by t test (P<0.05).

    Results

    The expression of specific megakaryocyte markers (CD41 and CD61) in the presence of different concentrations of MSC-MVs did not show any significant difference. Also, the expression of specific genes of megakaryocyte lineage was compared with control group. The expression of GATA2 and c-Mpl was significantly increased, GATA1 was not significantly decreased, and FLI1 was significantly decreased.

    Conclusion

    MSC-MVs could improve the expression of specific megakaryocyte genes; however, there was no significant expression of CD markers. Further studies, including the evaluation of late stages of megakaryocyte differentiation, are required to evaluate platelet production and shedding

    Keywords: Microvesicle, Hematopoietic stem, progenitor cells, Mesenchymal stem cells, Megakaryocyte
  • Saba Malekian, Marveh Rahmati, Soyar Sari, Monireh Kazemimanesh, Raheleh Kheirbakhsh, Ahad Muhammadnejad, Saeid Amanpour* Pages 323-328
    Purpose

    Triple-negative breast cancer (TNBC) is specified by high vascularity and repetitious metastasis. Although several studies have indicated that angiogenesis has an important role in invasive breast cancer, a suitable model of TNBC that can show the exact onset of angiogenesis factors still needs to be developed. The purpose of this study is to determine the expression level of angiogenesis factors in different clinical stages of the 4T1 tumor as TNBC mouse model.

    Methods

    Twenty mice were injected by the 4T1 cell line, and four mice selected as healthy controls. Following by tumor induction, the mice were randomly put into four groups, each contains four mice. Once the tumor volume reached to the early stage (<100 mm3 ), intermediate stage (100-300 mm3 ), advanced stage (300-500 mm3 ), and end stage (>500 mm3 ), they were removed by surgery. Then, the expression levels of Hif1α, VEGFR1, and VEGFR2 genes, as well as tumor markers of VEGF, bFGF and CD31, were evaluated by qPCR and immunohistochemistry (IHC) respectively. The statistical analysis was done by SPSS version 16.

    Results

    TNBC tumors were confirmed and multi-foci metastasis in the lung were seen. The mRNA and protein expression levels of the angiogenesis factors increased in the early stage and as the tumor grew, their expression level enhanced dramatically.

    Conclusion

    The 4T1 syngeneic mouse tumor may serve as an appropriate TNBC model for further investigation of the angiogenesis and therapies. Moreover, angiogenesis factors are induced before the advanced stage, and anti-angiogenesis therapy is necessary to be considered at the first line of treatment in TBNC.

    Keywords: Angiogenesis factor, bFGF, CD31, Triple-negative breast cancer, VEGF, VEGFR1, VEGFR2
  • Ahmad Shekari, Mehdi Forouzesh*, Roohollah Valipour, Fardin Fallah, Pardis Shojaei Pages 329-333
    Purpose

    We investigated validation and optimization of ultrasound-assisted dispersive liquidliquid microextraction (UADLLME) as a preparation method for detection of methadone in saliva samples.

    Methods

    We used blank and methadone-containing saliva samples and also standard methadone solution. Sodium hydroxide and chloroform were added to samples and they were held in ultrasonic bath. Then preparations were centrifuged and extracted analyte was analyzed by gas chromatography-mass spectrometry (GC-MS). Accuracy was measured by Intra and between-day mean relative errors (RE). Precision was assessed by coefficient of variation (CV). Recovery, specificity, linearity and limits of detection and quantification were also determined. Optimization was conducted for ultrasound duration, pH and extraction phase volume. Efficiency of dispersive liquid-liquid microextraction (DLLME) and UADLLME were compared.

    Results

    Intra and between-day accuracies (2.3 -7.5%), recovery (89.4-115.5%) and precision (5.2-11.3%) were all acceptable. Calibration curve was linear in the concentration range of 150 ng/mL-10 µL/mL with R2 >0.9995 and equation of y=86.901x-5342.5. Limits of detection and quantification were 50 and 150 ng/mL, respectively. Specificity was measured by comparing retention times of saliva samples (containing methadone metabolites and other commonly used drugs) during UADLLME/GC-MS analysis and no interference was observed. Recovery of UADLLME was 1.4 of DLLME. Solvent and sample volumes required for UADLLME were 1/200 and 1/20 of DLLME. The greatest efficiency obtained at pH of 10, with ultrasound treatment duration of 5 minutes and extraction phase volume of 1000 µL.

    Conclusion

    Study found that UADLLME/GC-MS is a valid and efficient method for detection of methadone in oral fluid.

    Keywords: Dispersive liquid-liquidmicroextraction, Gas chromatography-massspectrometry, Ultrasound, Methadone, Saliva, Validation