فهرست مطالب

Advanced Pharmaceutical Bulletin - Volume:9 Issue:4, 2019
  • Volume:9 Issue:4, 2019
  • تاریخ انتشار: 1398/08/28
  • تعداد عناوین: 20
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  • Sbonelo Khanyile, Priscilla Masamba, Babatunji Emmanuel Oyinloye, Londiwe Simphiwe Mbatha, Abidemi Paul Kapp Pages 510-520

    Chlorotoxin (CTX) is a minute 4 kDa protein made up of 36 amino acid residues, commonlyknown for its binding affinity to chloride channels and matrix metalloproteinase-2 (MMP-2)of glioma tumors of the spine and brain. This property and the possibility of conjugating thispeptide to nanoparticles have enabled its diverse use in various biotechnological and biomedicalapplications for cancer treatment, such as in tumor imaging and radiotherapy. Because of thefascinating biological properties CTX possesses, elucidating its mechanism of action may holdpromise for the development of new and effective therapeutic drugs, as well as more sensitiveand highly specific cancer-screening kits. This article therefore reviews the currently knownapplications of CTX and suggests diverse ways in which it can be applied for the design ofimproved drugs and diagnostic tools for cancer.

    Keywords: Cancer, Chlorotoxin, Diagnostics, Matrix metalloproteinase, Therapeutics, Tumor
  • Someshwar Komati*, Suryakanta Swain, Muddana Eswara Bhanoji Rao, Bikash Ranjan Jena, Vishali Dasi Pages 521-538

    Innovations in pharmaceutical research are striving for designing newer drug therapies toeradicate deadly diseases. Strategies for such inventions always flourish with keys and objectivesof minimal adverse effects and effective treatment. Recent trends in pharmaceutical technologyspecify that mucoadhesive drug delivery system is particularly appropriate than oral controlrelease, for getting local systematic delivery of drugs in GIT for an extended interval of time ata predetermined rate. However, it is somehow expensive and unpleasant sensation for somepatients, but still it is needful for getting short enzymatic activity, simple administration withoutpain and evasion of fast pass metabolism. Usually the vehicles employed in drug delivery ofmucoadhesive system have a significant impact that draws further attention to potential benefitslike improved bioavailability of therapeutic agents, extensive drug residence time at the site ofadministration and a comparatively faster drug uptake into the systemic circulation. The drugrelease from mucoadhesive multiparticulates is contingent on several types of factors comprisingcarrier need to produce the multiparticles and quantity of medication drug contained in them.Mucoadhesion is characterized by selected theories and mechanisms. Various strategiesemergent in mucoadhesive multiparticulate drug delivery system (MMDDS) by in-vitro as wellas ex-vivo description and characterization are also critically discussed. Apart from these, theprimary focus during this review is to highlight current patents, clinical status, and regulatorypolicy for enhancement of mucoadhesive multi-particulate drug delivery system in the present scenario.

    Keywords: Mucoadhesive materials, Multiparticulate systems, In-vitro, in-vivo methods, Isolated loop technique, Updated patents
  • Faramarz Rahmatizadeh, Shiva Gholizadeh, Ghaleh Aziz, Khodadad Khodadadi, Maryam Lale Ataei, Esmaeil Ebrahimie, Jafar Soleimani Rad*, Maryam Pashaiasl Pages 539-558

    Cancer has long been considered as a heterogeneous population of uncontrolled proliferation ofdifferent transformed cell types. The recent findings concerning tumorigeneses have highlightedthe fact that tumors can progress through tight relationships among tumor cells, cellular, andnon-cellular components which are present within tumor tissues. In recent years, studies haveshown that mesenchymal stem cells (MSCs) are essential components of non-tumor cells withinthe tumor tissues that can strongly affect tumor development. Several forms of MSCs have beenidentified within tumor stroma. Naïve (innate) mesenchymal stem cells (N-MSCs) derived fromdifferent sources are mostly recruited into the tumor stroma. N-MSCs exert dual and divergenteffects on tumor growth through different conditions and factors such as toll-like receptorpriming (TLR-priming), which is the primary underlying causes of opposite effects. Moreover,MSCs also have the contrary effects by various molecular mechanisms relying on direct cellto-cell connections and indirect communications through the autocrine, paracrine routes, andtumor microenvironment (TME).Overall, cell-based therapies will hold great promise to provide novel anticancer treatments.However, the application of intact MSCs in cancer treatment can theoretically cause adverseclinical outcomes. It is essential that to extensively analysis the effective factors and conditionsin which underlying mechanisms are adopted by MSCs when encounter with cancer.The aim is to review the cellular and molecular mechanisms underlying the dual effects ofMSCs followed by the importance of polarization of MSCs through priming of TLRs.

    Keywords: Mesenchymal stem cells, Dual effects, Bidirectional effects, Anti-tumor, Cell-cell interactions, Secretory factors, Tumor progression
  • Department of Pharmaceutics, Faculty of Pharmacy, Kuwait University, Health Sciences Center, Gabriya, Kuwait. Pages 559-570
    Purpose

    To enhance the dissolution rate of the poorly soluble drug atorvastatin calcium (ATC) bycocrystallization with selected coformers. Enhancement of the dissolution rate and solubility of thedrug, which is classified as Class II of the Biopharmaceutical Classification System (BCS), is expectedto enhance the bioavailability.

    Methods

    Two methods were used for preparing the cocrystals, solvent drop grinding (SDG) andsolvent evaporation (SE) method using 1:1, 1:3, and 1:10 drug-coformer molar ratios. Glucosaminehydrochloride (GluN) and nicotinamide (NIC) were investigated as coformers. The cocrystals,their physical mixtures, and the raw ATC were characterized by fourier transform infrared (FTIRspectroscopy), differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD), massspectroscopy (MS), scanning electron microscopy (SEM), solubility, and dissolution rate studies.

    Results

    SDG and SE were effective in improving the dissolution rate of ATC with both coformers.Drug: coformer ratio 1:3 was optimum. The solubility values for ATC, GluN-, and NIC-cocrystals were26, to 35 and 50 μg/mL, respectively. The dissolution rate of ATC from cocrystals was > 90% after 5minutes, compared to 41% untreated ATC.

    Conclusion

    Cocrystallization significantly improved the solubility and dissolution, in comparison tothe untreated ATC.

    Keywords: Atorvastatin, Glucosamine, Nicotinamide, Cocrystallization, Characterization
  • Ali Darini, Touba Eslaminejad, Seyed Noroddin Nematollahi Mahani, Mehdi Ansari* Pages 571-577
    Purpose

    The main aim of the present study was to design, fabrication and physicochemicalcharacteristics of the magnetogel nanospheres as carriers for Cisplatin in the in vitro environment.

    Methods

    Magnetic nanospheres were synthesized by using a chemical co-precipitation methodand coated by sodium alginate through double emulsion method. Then cisplatin was encapsulatedinto β-cyclodextrin -sodium alginate grafted magnetic nanospheres. The physicochemicalproperties of the sodium alginate grafted magnetic nanospheres were characterized by using FTIR,particle size analyzing, vibrating sample magnetometry, thermogravimetric and SEM analysis.Also the drug entrapment efficiency, content and in vitro release profile were investigated.

    Results

    Size distribution results revealed that the particles size was distributed in the rangeof 50± nm. Also morphological properties showed that particles are separated and sphericalwith the grafted layers of the polymer. The release profile data were in the acceptable rangecompared to the blank (cisplatin solution).

    Conclusion

    It could be concluded that the sodium alginate grafted magnetic nanospheres couldact as a slow and controlled release system to deliver cisplatin.

    Keywords: Alginate, Beta-cyclodextrin, Cancer, Cisplatin, Controlled release, Drug delivery systems, Magnetic nanoparticles
  • Saman Mohammadi, Abbas Pardakhty, Maryam Khalili, Reza Fathi, Maryam Rezaeizadeh, Saeedeh Farajzadeh, Azadeh Mohebbi, Mahin Aflatoonian Pages 578-583
    Purpose

    Combination of benzoyl peroxide (BPO) with topical antibiotics can lead to higherefficacy and less bacterial resistance, but it in turn increases adverse effects such as skinirritability and dryness. In this study, the efficacy of combination therapy of niosomal BPO 1%and clindamycin (CL) 1% is compared with niosomal CL in acne vulgaris.

    Methods

    This is a double-blind clinical trial study on 100 patients with acne vulgaris inAfzalipour hospital in Kerman. Patients were randomly divided into 2 groups (case and control).The case group received niosomal combination of BPO 1% and CL 1%.The control groupreceived niosomal CL1%. The efficacy of treatment protocols was evaluated in 2nd, 4th, 8thand 12th weeks of treatment by counting lesions (severity and grading acne lesions) and qualityof life (QoL). Furthermore, side effect were evaluated at each treatment visits.

    Results

    The reduction in mean percentage of acne lesions in case group (treated with BPO 1%and CL1%) (64.21%) was higher than control group (treated with niosomal CL 1%) (59.04%),but the statistical difference was not significant. Sum of excellent and good results were foundin 80% and 76.1% of case and control groups, respectively (P = 0.377). Also adding BPO to thetreatment formulation in case group did not increase adverse effects, as statistical differencebetween 2 groups was not significant.

    Conclusion

    Combination of niosomal BPO 1% and CL 1% in treatment of acne vulgaris showedhigher efficacy with no increase in adverse effects in comparison with niosomal CL 1%, but thestatistical difference was not significant.

    Keywords: Acne, Benzoyl peroxide, Clindamycin, Niosome
  • Saeed Mohammad Soleymani, Anayatollah Salimi* Pages 584-592
    Purpose

    Finasteride is a pharmaceutical agent that treats hair loss and acne with hormonalpatterns. Due to its poor water solubility, and the smaller surface area in comparison to totalskin surface area, penetration of the drug into hair follicles and skin is low. The aim of thisresearch was to formulate, characterize and evaluate in vitro skin permeability of finasteridemicroemulsions (MEs).

    Methods

    Finasteride MEs were prepared using a pseudo-ternary phase diagram method withan appropriate ratio of oil mixture, surfactant-co-surfactant mixture and water. MEs containing1% finasteride were prepared with a suitable amount of oily phase and surfactant and cosurfactant.The physicochemical properties of these MEs and in vitro skin permeability of MEswere evaluated.

    Results

    The results showed that the mean droplet size range of ME samples was 5–17 nm andpH was 5.1–5.7. The viscosity of MEs ranged from 86.4–209.6 cps. The drug release profileshowed that 49.510% of the drug was released (ME-F-6) over the 24 hours of the experiment.The kinetics of drug release from all selected MEs were approximately described by Higuchiand first-order modeling. All ME formulations with different compositions and propertiessignificantly increased flux and permeability coefficient from rat skin. The selected MEs exhibit99.9% finasteride after six months of storage.

    Conclusion

    This study showed that any change in the content and composition of MEs couldchange the physical and chemical properties in addition to ME permeability parameters. TheMEs increased permeability of the skin to finasteride.

    Keywords: Dermal drug delivery, Finasteride, Microemulsion, Permeability, Release
  • Ghobad Mohammadi, Shahla Mirzaeei*, Shiva Taghe, Pardis Mohammadi Pages 593-600
    Purpose

    The purpose of the present study was to improve the ocular delivery for ketorolactromethamine (KT) used to treat inflammation of the eye.

    Methods

    Eudragit nanoparticles loaded with KT were prepared and incorporated in polyvinylalcohol (PVA) and hydroxyethyl cellulose (HEC) films. Nanoparticles were characterized byFourier transform-infrared (FT-IR), scanning electron microscopy (SEM). Physicochemicalproperties and encapsulation effciency were investigated for nanoparticles. Also, the insertswere evaluated for their physiochemical parameters like percentage moisture absorption,percentage moisture loss, thickness and folding endurance.

    Results

    Mean particle size and zeta potential were in range of 153.8-217 nm and (-10.8) -(-40.7) mV, respectively. The results show that the use of a surfactant has not led to any majorchange on drug loading. The loading increases with the amount of polymer. The insert had athickness varying from 0.072 ± 0.0098 to 0.0865 ± 0.0035 mm. The thicknesses of the insertsand the folding endurance increased with the total polymer concentration. The physicochemicalproperties showed that the Eudragit® L-100 nanoparticles loaded PVA-HEC films could be aneffective carrier for KT.

    Conclusion

    For the first time, inserts of Eudragit nanoparticles were successfully prepared forophthalmic drug delivery system to prevent frequent drug administration and enhance patientcompliance.

    Keywords: Ketorolac tromethamine, Inserts of Eudragit® L100 nanoparticles, Ophthalmic drug delivery, PVA
  • Hamid Rahmani, Ali Fattahi*, Komail Sadrjavadi, Salar Khaledian, Yalda Shokoohinia Pages 601-608
    Purpose

    The aim of this study is to prepare 5-fluorouracil (5-FU) loaded silk fibroin nanoparticles(SFNPs) and to achieve a controlled release delivery system with the high loading capacity.

    Methods

    SFNPs with 1:1, 1:3, and 1:10 ratios of 5-FU to silk fibroin were prepared. SFNPswere characterized by Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD)analysis, Scanning electron microscope (SEM), and Transmission electron microscope (TEM).Loading efficiency, in vitro release, and cell viability were studied for optimal SFNPs.

    Results

    The ratio of 1:1 was optimal formulation with the size and polydispersity index (PDI)of 221.03 nm and 0.093 before freeze drying, and 286.7 nm and 0.154 after freeze dryingby lactose, respectively. The loading efficiency and loading content of this ratio were 52.32%and 34.35%, respectively. FT-IR and XRD analysis indicated the conformational change (fromrandom coil to β-sheet) in the structure of nanoparticles by increasing amount of the drug, whichcaused the smaller size, the higher loading efficiency, and the slower release pattern. The drugloadednanoparticles reached to the half maximal inhibitory concentration (IC50) that werecomparable with free drug on MCF7 (human breast cancer) cell line.

    Conclusion

    This study was planned to achieve a promising controlled release drug deliverysystem for carrying 5-FU, as a potent anticancer drug. SFNPs were found proper candidates fordelivery of a hydrophilic drug such as 5-FU.

    Keywords: SF, 5-Fluorouracil, Nanoparticle, Precipitation, Cytotoxicity
  • Ilham Kuncahyo, Syaiful Choiri*, Achmad Fudholi, Ronny Martien, Abdul Rohman Pages 609-618
    Purpose

    Recently, a self-nanoemulsifying drug delivery system (SNEDDS) has showngreat improvement in the enhancement of drug bioavailability. The selection of appropriatecompositions in the SNEDDS formulation is the fundamental step towards developing asuccessful formulation. This study sought to evaluate the effectiveness of fractional factorialdesign (FFD) in the selection and screening of a SNEDDS composition. Furthermore, the mostefficient FFD approach would be applied to the selection of SNEDDS components.

    Methods

    The types of oil, surfactant, co-surfactant, and their concentrations were selected asfactors. 26 full factorial design (FD) (64 runs), 26-1 FFD (32 runs), 26-2 FFD (16 runs), and 26-3 FFD(8 runs) were compared to the main effect contributions of each design. Ca-pitavastatin (Ca-PVT)was used as a drug model. Screening parameters, such as transmittance, emulsification time,and drug load, were selected as responses followed by particle size along with zeta potentialfor optimized formulation.

    Results

    The results indicated that the patterns of 26 full FD and 26-1 for both main effects andinteractions were similar. 26-3 FFD lacked adequate precision when used for screening owing tothe limitation of design points. In addition, capryol, Tween 80, and transcutol P were selected tobe developed in a SNEDDS formulation with a particle size of 69.7 ± 5.3 nm along with a zetapotential of 33.4 ± 2.1 mV.

    Conclusion

    Herein, 26-2 FFD was chosen as the most efficient and adequate design for theselection and screening of SNEDDS composition. The optimized formulation fulfilled therequirement of a quality target profile of a nanoemulsion.

    Keywords: Fractional factorial design, SNEDDS, Screening, Optimization, Statistical approach
  • Wira Eka Putra, Muhaimin Rifa’i* Pages 619-623
    Purpose

    Accumulating evidence shows the genus of Sambucus exerts a broad spectrum ofmedicinal potencies such as anticancer, antiviral, antibacterial, and antidiabetes. Based on theprevious studies, we hypothesized that bioactive compounds of Sambucus might alter severalbiological systems, including the immune system. Therefore, this study extensively aimed toevaluate the immunomodulatory activities of Sambucus javanica extracts in 7,12-dimethylbenz[a]anthracene (DMBA)-treated BALB/c mouse.

    Methods

    The experimental mice were orally administrated with 2.8 mg.kg-1 BW of DMBA forten times within a month. After that, the mice were treated by S. javanica berries and leavesextracts for 2 weeks. Subsequently, the inflammation rate was evaluated by using flow cytometryanalysis, whereas the necrosis incidences were observed by hematoxylin & eosin staining.

    Results

    Based on the results, we found the expression of tumor necrosis factor alpha (TNF-α)and interferon gamma (IFN-ɣ) were increased however after treated by S. javanica berries andleaves extracts were significantly decreased. In the same way, necrosis incidence was increasedin the DMBA-treated group however it was diminished with S. javanica extracts treatment.

    Conclusion

    Together, these results suggested that S. javanica extracts have immunomodulatoryactivities to suppress inflammation and reduce necrosis incidence in experimental mice.

    Keywords: DMBA, BALB, c mice, Immunomodulation, Inflammation, Sambucus javanica
  • Sepideh Nikfarjam, Mohammad Reza Tohidkia*, Tayebeh Mehdipour, Ramin Soleimani, Ali Akbar Rahim Rahimi, Mohammad Nouri Pages 624-631
    Purpose

    Generation of antibodies which potentially discriminate between malignant andhealthy cells is an important prerequisite for early diagnosis and treatment of gastric cancer(GC). Comparative analysis of cell surface protein landscape will provide an experimental basisfor biomarker discovery, which is essential for targeted molecular therapies. This study aimedto isolate phage-displayed antibody fragments recognizing cell surface proteins, which weredifferentially expressed between two closely related GC cell lines, namely AGS and MKN-45.

    Methods

    We selected and screened a semisynthetic phage-scFv library on AGS, MKN-45, andNIH-3T3 cell lines by utilizing a tailored selection scheme that was designed to isolate phagescFvsthat not only recognize the differentiated AGS cells but also distinguish them from NIH-3T3 fibroblasts and the poorly differentiated MKN-45 cells.

    Results

    After four rounds of subtractive whole cell panning, 14 unique clones were identifiedby ELISA screening and nucleotide sequencing. For further characterization, we focused on fourphage-scFvs with strong signals in screening, and their specificity was confirmed by cell-basedELISA. Furthermore, the selected phage-scFvs were able to specifically stain AGS cells with38.74% (H1), 11.04% (D11), 76.93% (G11), and 69.03% (D1) in flow cytometry analysis whichsupported the ability of theses phage scFvs in distinguishing AGS from MKN-45 and NIH-3T3cells.

    Conclusion

    Combined with other proteomic techniques, these phage-scFvs can be applied tomembrane proteome analysis and, subsequently, identification of novel tumor-related antigensmediating proliferation and differentiation of cells. Furthermore, such antibody fragments canbe exploited for diagnostic purposes as well as targeted drug delivery of GC.

    Keywords: Gastric cancer, Phage display, Proteomics, scFv, Tumor targeting, Whole cell panning
  • Esmaeil Abasi, Shahram Shahabi, Majid Golkar, Peyman Khezri, Habib Mohammadzadeh Hajipirloo* Pages 632-639
    Purpose

    Propranolol as a novel adjuvant, was used to evaluate the immunogenic effect of threedoses of recombinant SAG-1 (rSAG-1) antigen of Toxoplasma gondii in BALB/c mice for findingthe optimal dose, and was compared with efficacy of tachyzoite lysate antigen (TLA).

    Methods

    Eight different groups of 15 BALB/c mice received different volumes of theimmunogenic material (three doses of r SAG-1 and one dose of TLA antigens), with or withoutpropranolol adjuvant, subcutaneously. The control group mice received only PBS. Three weeksafter the last immunization, the serum levels of IgG2a, IgG1 and IgG total antibodies againstTLA, splenic interleukin-5 (IL-5) and Interferon-gamma (IFN-γ) (produced against TLA) and thesplenic lymphocyte proliferation after adding TLA were measured to evaluate humoral andcellular immune responses. Challenge test was performed by subcutaneously injection of 1000alive and active tachyzoites in to five mice per each group and survival days for each group ofmice were recorded.

    Results

    The mice group that received propranolol adjuvant and 20 μg of r SAG-1 antigenper dose of injection showed significantly more IFN-γ production, more proliferation ofsplenic lymphocytes and higher anti-TLA-specific IgG2a production (three main indexes forcell mediated immunity) in comparison with other groups. Moreover, in the challenge test,this group of mice had a significantly increased survival time, indicating the positive effect ofpropranolol in the more stimulating of cellular immunity that is necessary for toxoplasmosisprevention or suppress.

    Conclusion

    Our results showed that T. gondii rSAG-1 antigen in combination with propranololas adjuvant (which can induce Th1 related responses) are good candidates for further study toa vaccine design.

    Keywords: Adjuvant, Propranolol, r SAG-1, Toxoplasma gondii, Vaccine
  • Sayed Sharif Balkhi, Zohreh Hojati* Pages 640-648
    Purpose

    Interferon beta (IFN-β) is used to combat multiple sclerosis (MS) disease. CreatingR27T and V101F mutations (mHuIFN-β-27 and mHuIFN-β-101) is one of the tasks performedto improve human interferon beta (HuIFN-β) half-life, function and expression. In this work,the impact of R27T and V101F mutations in recombinant IFN-β on its binding to interferonreceptors were studied by molecular docking.

    Methods

    This work was performed through in silico study. The simulation of mutation wasperformed using the online Rosetta Backrub software and checked using server verify3D.Comparison of access to the solvent of the amino acids in the structures created was performedusing the asaview online server. Also, the effect of mutations on the fold of the protein wasreviewed by the online HOPE server. The molecular docking was performed between HuIFN-βand the external region of IFNAR receptor using the online ClusPro2 protein-protein dockingserver.

    Results

    The comparison of the values of the negative binding energy (ΔGbind) obtained fromprotein-protein molecular docking between IFNAR receptor and HuIFN-β, mHuIFN-β-27,mHuIFN-β-101 and mHuIFN-β-27-101 ligands did not show a significant difference, and thesedifferences do not see any meaningful relationship between them (P > 0.9999).

    Conclusion

    Regarding these results, it can be concluded that these mutations do not have anegative effect on the composition of the complex rHuIFN-β/IFNAR. So, they do not interferewith the binding of the IFN-β to the receptor. It is concluded that the quality of the rHuIFN-β isimproved by introducing these two mutations.

    Keywords: Interferon Beta, Molecular Docking, Insilico, IFNAR, mHuIFN-β-27-101
  • Mansoureh Mirabdollahi, Shaghayegh Haghjooy Javanmard, Hojjat Sadeghi, Aliabadi* Pages 649-654
    Purpose

    Several attempts have been made to identify the mechanisms by which mesenchymalstem cells (MSCs)-derived secretome exert anti-tumor or tumorigenic effects, but still furtherinvestigations are needed to explore this subject. Thus, in this study we want to examine theexpression of different cytokines in secretome of hWJ-MSCs and their effects on cytokineexpression profile of the MCF-7 tumor cells.

    Methods

    The hWJ-MSCs were isolated and characterized according to the International Societyfor Cellular Therapy criteria. Then, secretome of hWJ-MSCs was collected and freeze-dried, and20 mg/mL of the freeze-dried secretome was used to treat MCF-7 cancer cells for 48 hours.Afterwards, the expression levels of 12 cytokines including IL-1a, IL-1b, IL-2, IL-4, IL-6, IL-8,IL-10, IL-12, IL-17A, TNFα, IFNγ and GM-CSF in secretome of hWJ-MSCs alone as well as insupernatant of tumor cells before and after treatment with hWJ-MSCs secretome were evaluated.

    Results

    Our results indicate that MCF-7 cells express significant amount of IL-6 and IL-8.Moreover, significant amounts of IL-1a, IL-1b, IL-8, IL-6 and GM-CSF were detected in secretomeof hWJ-MSCs. Furthermore, IL-1a, IL-2 and IL-4 were expressed significantly by MCF-7 cellsafter their treatment with hWJ-MSCs-derived secretome.

    Conclusion

    According to our findings, the hWJ-MSCs derived secretome contains differentcytokines which can exert either anti-tumor or tumorigenic effects.

    Keywords: Human Wharton’s Jelly mesenchymal stem cells (hWJ-MSCs), MCF-7, Tumor cells, Secretome, Cytokine expression profile
  • Ahmed Saeed Arafa, Amany Elsayed Ragab*, Abdel, Rahim Sayed Ibrahim, Wael Saad Abdel, Mageed, Mahmoud Emam Nasr Pages 655-661
    Purpose

    Strictosidine-β-D-glucosidase (SGD) is considered as a key enzyme in the productionof bisindole alkaloids in Catharanthus roseus. The present study illustrated the production of ashort sequence of this enzyme in Escherichia coli without codon optimization.

    Methods

    Strictosidine-β-D-glucosidase (sgd) gene short sequence (1434 bp), which lacksthe conserved sequence KGFFVWS and the localization peptide sequence at the C-terminal,was amplified from cDNA of C. roseus leaves, cloned and expressed in Escherichia coli. Theactivity of the produced protein in cell free lysate was tested using total alkaloid extract of C.roseus leaves.

    Results

    HPLC and LC-MS analysis of the assay mixture revealed the disappearance of thestrictosidine peak.

    Conclusion

    SGD short sequence can be produced in Escherichia coli in active form withoutcodon optimization.

    Keywords: Catharanthus roseus, Escherichia coli, Strictosidine-β-D-glucosidase, Enzyme assay, Overexpression
  • Faraz Sigaroodi, Hajar Shafaei*, Mohammad Karimipour, Mohammad Amin Dolatkhah, Abbas Delazar Pages 662-667
    Purpose

    Natural biomaterials are a key base in tissue engineering, and collagen, as the maincontent of the extracellular matrix (ECM), is frequently used in tissue engineering. Aloe verahas some therapeutic effects on ulcers, therefore, the use of this natural resource has alwaysbeen considered for improving collagen function. We aimed to evaluate the effect of Aloe vera/Collagen blended on cell viability, cell attachment, and angiogenic potential by determining ofintegrin α1β1 and platelet endothelial cell adhesion molecule (PECAM-1) genes expression inhuman adipose-derived stem cells (hASCs).

    Methods

    In this study, hASCs after harvesting of adipose tissues from abdominal subcutaneousadipose tissue and isolation, were cultured in four groups of control, collagen gel, Aloe veragel, and Aloe vera/collagen blended in vitro environment at 24h and then cell viability wasassessed by MTT (3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium) assay. Integrin α1β1 andPECAM-1 genes expression were evaluated by real-time RT-PCR.

    Results

    The results of MTT showed that the combination of Aloe vera/collagen was retained thecell viability at the normal range and improved it. In real-time RT-PCR results, integrin α1β1 andPECAM-1 gene expression were increased in the Aloe vera/collagen blended group comparedto the control group.

    Conclusion

    For tissue engineering purposes, Aloe vera improves collagen properties in theculture of hASCs by increasing the expression of the integrin α1β1 and PECAM-1 genes.

    Keywords: Adipose-derived stem cells, Aloe vera gel, Collagen, Integrin, PECAM-1, Tissue engineering
  • Haniyeh Abuei, Abbas Behzad, Behbahani, Fatemeh Faghihi, Ali Farhadi*, Gholam Reza Rafiei Dehbidi, Mohammad Pirouzfar, Farahnaz Zare Pages 668-673
    Purpose

    Despite all the efforts for discovery of efficient anti-cancer therapeutics, cancer is stilla major health concern worldwide. p28 is a bacterial small peptide which has been widelyinvestigated due to its preferential cell internalization and anti-cancer activities. Intracellularly,p28 offers its anti-cancer traits by impeding the degradation of tumor-suppressor protein "p53".In this study, we investigated the potency of p28 in inducing apoptosis or decreasing cellviability in p53-null "HeLa" cell line.

    Methods

    The coding sequence for p28 peptide was obtained from Pseudomonas aeruginosaby PCR amplification of the p28 gene. The coding gene was cloned in pET-28a vector andtransformed into E. coli bacterial host. Subsequently, the expressed peptide was purified usingNi-NTA chromatography system and introduced into the target cells. The anti-proliferative andapoptotic activity of p28 on HeLa and HEK-293 cells were investigated using MTT and PEAnnexinV Flowcytometry assays.

    Results

    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Westernblotting confirmed the expression of p28 peptide in the bacterial host. Bradford assay revealeda concentration of 0.05 mg/mL for the purified p28. MTT assay of cells treated with p28 atconcentrations of 0, 0.5, 1, 2 and 2.5 μM indicated 24h viability values of 97%, 89%, 88%,87% and 84% for HeLa cells, respectively. Data obtained from flowcytometry analyses revealed24h apoptosis rate of 7.17%, 8.05%, 8.63% and 8.84% for HeLa cells treated with 0, 0.5, 1,and 2 μM p28, respectively.

    Conclusion

    MTT and flowcytometry apoptosis assays suggest no statistically significant effectof p28 on the viability and apoptosis status of p53-null HeLa cells when results compared todata obtained from HEK-293 cells (P > 0.05). These results imply that anti-cancer efficacy of p28is directly dependent on the presence of p53, suggesting p28 as an inappropriate therapeuticagent for treatment of cancers with negative p53 status.

    Keywords: p28, p53, Null HeLa cells, Apoptosis, Viability
  • Nail Salavatovich Akhmadiev, Albina Midkhatovna Galimova, Vnira Rakhimovna Akhmetova*, Veronika Radievna Khairullina, Rozaliia Akramovna Galimova, Eduard Feliksovich Agletdino, Askhat Gabdrahmanovich Ibragimov, Valery Alekseevich Kataev Pages 674-684
    Purpose

    In order to investigate mechanisms underlying the hepatoprotective action of S,Spalladaheterocycle,inhibition of cytochromes P450 has been modeled by molecular dockingof four palladaheterocycle stereoisomers to the active sites of an enzymatic oxidase system. Toobtain a deeper insight into biochemical aspects providing a basis for the therapeutic effects offive-membered palladacycles (as mixture of stereoisomers), a number of preclinical trials hasbeen conducted

    Methods

    2D and 3D structures of palladaheterocycle stereoisomers were obtained viaconverting into SDF files by means of software MarvinSketch. Binding of palladaheterocycle atthe active sites of cytochromes P450 2E1 and P450 2C9 has been studied by molecular dockingusing LeadIT 2.3.2. Hepatoprotective activity of palladaheterocycle at 2.5, 25 and 250 mg/kgdoses has been studied based on a model of acute intoxication by CCl4 using in vivo methods.

    Results

    By molecular docking it was identify amino acid fragments responsible for bindingwith palladacyclic isomers. The tested compound is comparable, in terms of its activity tothe hepatoprotective drug SAM according to the in vivo and in vitro experiments such asanimal survival data, the efficiency of correction of the cytolytic syndrome, the liver excretoryfunction, carbohydrate, protein and lipid metabolism, and the correction efficiency of the liverantitoxic function (the latter has been determined based on the results of a hexobarbital controlexperiment).

    Conclusion

    Taking into account results obtained in vivo, in vitro and in silico, it can be concludedthat the five-membered S,S-palladaheterocycle effectively protect the liver against acute damagecaused by CCl4, via activation of catalase and glucuronyltransferase, as well as via inhibition ofthe oxidative stress enzymes.

    Keywords: Cytochrome P450 CYP2E1, Hepatoprotective activity, Liver, Molecular docking simulation, Palladaheterocycle, Preclinical study
  • Andrea Rasmussen, Kaylee Murphy, David W. Hoskin* Pages 685-689
    Purpose

    Gingerol homologs found in the rhizomes of ginger plants have the potential to benefithuman health, including the prevention and treatment of cancer. This study evaluated the effectof 10-gingerol on ovarian cancer cell (HEY, OVCAR3, and SKOV-3) growth.

    Methods

    Cell growth was measured by MTT assays, flow cytometry was used to assess cellproliferation, cytotoxicity and cell cycle progression, and western blotting was used to measurecyclin protein expression.

    Results

    Ovarian cancer cells that were treated with 10-gingerol experienced a time- anddose-dependent decrease in cell number, which was due to a reduction in cell proliferationrather than a cytotoxic effect. Reduced proliferation of 10-gingerol-treated ovarian cancercells was associated with an increased percentage of cells in G2 phase of the cell cycle anda corresponding reduction in the percentage of cells in G1. Ovarian cancer cells also showeddecreased cyclin A, B1, and D3 expression following exposure to 10-gingerol.

    Conclusion

    These findings revealed that 10-gingerol caused a G2 arrest-associated suppressionof ovarian cancer cell growth, which may be exploited in the management of ovarian cancer.

    Keywords: Cell cycle, Proliferation, Ginger, 10-Gingerol, Ovarian cancer