فهرست مطالب

Advanced Pharmaceutical Bulletin - Volume:11 Issue: 1, Jan 2021

Advanced Pharmaceutical Bulletin
Volume:11 Issue: 1, Jan 2021

  • تاریخ انتشار: 1399/09/01
  • تعداد عناوین: 24
|
  • Aziz Homayouni Rad, Leili Aghebati Maleki, Hossein Samadi Kafil, Hamideh Fathi Zavoshti, Amin Abbasi* Pages 1-5

    As many investigations have reported, there is a complicated relation between fermented foods, lactic acid bacteria (LAB), and human health. It seems that bioactive components such as prebiotics, probiotics, and postbiotics are key mediators of the complex and direct association between these factors. LAB activity in the matrix of fermented foods and improving their growth by prebiotic compounds ultimately results in the production of bioactive molecules (postbiotics), which possess specific biological and physiological properties. The term "postbiotics" refers to a complex of biological micro- and macromolecules, if consumed in adequate amounts, provides the host with different health-promoting effects. Different reports have suggested that postbiotics possess the ability to moderate the effectiveness of cancer treatment and reduce the side-effects of conventional therapies in cancer patients due to their anti-proliferative, anti-inflammatory and anti-cancer properties. Consequently, postbiotics, for their unique characteristics, have gained great scientific attention and are considered as a novel approach for adjuvant therapy in patients with cancer.

    Keywords: Cancer, Postbiotic, Probiotic, Health, Treatment
  • Javad Mahmoudi, Saeed Sadigh-Eteghad, Hanieh Salehi-Pourmehr, Afshin Gharekhani, Mojtaba Ziaee * Pages 6-7
  • Mehdi Pasalar, Babak Daneshfard *, Kamran Bagheri Lankarani Pages 8-9
  • Arti Gupta *, Sonia Pandey, Jitendra Singh Yadav Pages 10-27

    Tuberculosis (TB) is a contagious disease that has affected mankind. The anti-TB treatment has been used from ancient times to control symptoms of this disease but these medications produced some serious side effects. Herbal products have been successfully used for the treatment of TB. Gold is the most biocompatible metal among all available for biomedical purposes so Gold nanoparticles (GNPs) have sought attention as an attractive biosynthesized drug to be studied in recent years for bioscience research. GNPs are used as better catalysts and due to unique small size, physical resemblance to physiological molecules, biocompatibility and non-cytotoxicity extensively used for various applications including drug and gene delivery. Greenly synthesized GNPs have much more potential in different fields because phytoconstituents used in GNP synthesis itself act as reducing and capping agents and produced more stabilized GNPs. This review is devoted to a discussion on GNPs synthesis with herbs for TB. The main focus is on the role of the natural plant bio-molecules involved in the bioreduction of metal salts during the GNPs synthesis with phytoconstituents used as antitubercular agents.

    Keywords: Green Synthesis, Gold nanoparticles, Tuberculosis, Phytoconstituents
  • Saba Mehrandish, Shahla Mirzaeei* Pages 28-38

    Ocular fungal infections affect more than one million people annually worldwide. They can lead to impaired vision or even complete blindness, so they should be treated immediately to prevent such consequences. Although topical administration has always been the most common route of ocular drug delivery owing to high patient acceptance, reduced side effects, and the possibility of self-administration, its limited ocular bioavailability poses a major challenge. As a result, attention has recently been drawn to the design and development of novel drug delivery systems (NDDS) that can overcome the challenges of conventional dosage forms. This research is the first to review and classify the studies which have designed and developed topical ocular NDDS with the aim to compare the performance and antifungal activity of these novel systems with conventional forms. According to the results, all studies seemed to confirm the superiority of NDDS over conventional forms in cases of released and permeated drug and antifungal activity. The NDDS were used specifically to improve ocular delivery by slowing down the release rate, increasing drug permeation, and subsequently increasing the antifungal effects of the active pharmaceutical ingredients. Hence, further studies on NDDS may aid the optimization of ocular drug delivery of antifungal drugs.

    Keywords: Antifungal drugs, Conventional dosage forms, Novel drug delivery systems, Ocular drug delivery
  • Mahnaz Nazari, Ehsan Javandoost, Mehdi Talebi, Aliakbar Movassaghpour *, Masoud Soleimani Pages 39-55

    Platelet-derived microparticles (PMPs) are a group of micrometer-scale extracellular vesicles released by platelets upon activation that are responsible for the majority of microvesicles found in plasma. PMPs’ physiological properties and functions have long been investigated by researchers. In this regard, a noticeable area of studies has been devoted to evaluating the potential roles and effects of PMPs on cancer progression. Clinical and experimental evidence conflictingly implicates supportive and suppressive functions for PMPs regarding cancer. Many of these functions could be deemed as a cornerstone for future considerations of PMPs usage in cancer targeted therapy. This review discusses what is currently known about PMPs and provides insights for new and possible research directions for further grasping the intricate interplay between PMPs and cancer.

    Keywords: PMP, Platelet, Microparticle, Cancer
  • Mohammed M. Mehanna*, Amina Tarek Mneimneh Pages 56-67

    The drug delivery investigation field is continuously widened and adapted to overcome many factors such as poor drug solubility, absorption, rapid metabolism, the variability of drug plasma levels, cellular efflux and many others. Due to resemblance to body constituents and their biocompatibility, lipids offer a promising scheme for poorly water-soluble and lipophilic drugs. Various nanoparticles including vesicular systems, lipid particulate systems, and emulsion systems provide some unique benefits as pharmaceutical carriers in drug and biomolecules delivery systems. Nowadays synthesis is directed toward simple, costless techniques, therefore, self-emulsifying systems have gained superiority over the other carriers. Self nano-emulsifying systems composed of oil, surfactant, and co-surfactant emulsified upon contact with an aqueous medium, has been widely exploited. This review attempts to provide a comprehensive interpretation of different types of lipid-based carriers emphasizing on the self-nanoemulsifying system, why it is gaining interest, formulation, composition, and applications.

    Keywords: Drug delivery, Lipids, Solubility, Self-nanoemulsifying system
  • Mohamed Fathallah Abou El-Nour, Sayed Hamed Kenawy, Gehan T. El-Bassyouni, Esmat Mahmoud Aly Hamzawy * Pages 68-76
    Purpose

    Praziquantel (PZQ) is a well-known drug accredited by the World Health Organization (WHO) for the treatment of schistosomiasis. It shows poor efficiency in patients during the earliest infection phases. Therefore, the search for new alternative drugs was the intention of many researchers.

    Methods

    In the current study, the effect of different concentrations (ranging from 0.07-10 μg∕mL) of calcium silicate (CS) containing 5% copper oxide [CS-5%CuO] on golden hamster infected by Schistosoma mansoni and Schistosoma haematobium (Egyptian strains) was evaluated in both in vitro and in vivo. To the best of our knowledge, this is a novel study in investigating the efficiency of CS-5%CuO against both strains of schistosomes. The worms of S. mansoni and S. haematobium were tested in RPMI-1640 medium in vitro.

    Results

    The results declare that CS-5% CuO exhibited excellent anti-schistosomal activities on both in vitro and in vivo experiments for both Egyptians Schistosoma strains. The most potential effect of the CS-5% CuO was exhibited after 6 h by 10 μg∕mL with significant activity of (P value = 0.001).

    Conclusion

    Therefore, CS-5%CuO may become an innovative treatment for the schistosomiasis.

    Keywords: Anti-schistosomal activity, CS-5%CuO-containing calcium silicate, S. mansoni, S. haematobium
  • Wawaimuli Arozal, Melva Louisa, Deni Rahmat, Priska Chendrana, Ni Made Dwi Sandhiutami * Pages 77-85
    Purpose

    This study aimed to provide the method of preparation, characterization of curcumin-loaded chitosan-sodium tripolyphosphate (NaTPP) nanoparticle, and evaluate its pharmacokinetic profiles.

    Methods

    Curcumin-loaded chitosan-NaTPP nanoparticles were synthesized using ionic gelation methods. Curcumin was dissolved using surfactants and cosurfactants. Chitosan polymer was then mixed in the curcumin solution and dripped with NaTPP solution until nanoparticle formation. The mucoadhesive study was evaluated by measuring the fluorescence of curcumin within the prepared nanoparticles. The pharmacokinetic profiles of curcumin particles and nanoparticles were then assessed in rats by administering a single oral dose of 100 mg/kg BW. Blood samples were taken from nine predetermined time points, and curcumin plasma concentrations were then analyzed using UPLC-MS/MS.

    Results

    The particle size of the curcumin nanoparticles obtained were 11.5 nm. Entrapment efficiency (EE) of curcumin nanoparticles were exceeding 99.97%, and drug loading capacity (DLC) was 11.34%. The mucoadhesive properties of the nanoparticles were superior to that of curcumin particles. Pharmacokinetic evaluation in rats revealed that curcumin nanoparticles resulted in an increase of area under the curve (AUC), maximum concentration (Cmax), earlier time to reach maximum concentration (Tmax), and lower clearance (CL).

    Conclusion

    Curcumin-loaded chitosan-NaTPP nanoparticles is an effective formulation to improve curcumin plasma concentrations. Thus, enable its applications for the treatment of various diseases.

    Keywords: Curcumin, Chitosan, Nanoparticles, Mucoadhesive, Pharmacokinetics
  • Asmaa Nabil Elboraey*, Hanan Hassan Abo-Almaged, Ahmed Abd El-Rahman El-Ashmawy, Aya Rashad Abdou, Amani Ramadan Moussa, Laila Hassanian Emara, Hossam Mohammed El-Masry, Gehan El-Tabie El Bassyouni, Magda Ismail Ramzy Pages 86-95
    Purpose

    This study aimed to evaluate the biological and mechanical properties of the poly(methyl methacrylate) (PMMA) denture base material as a vehicle incorporating novel hydroxyapatite nanoparticles (HA-NP) loaded with metronidazole (MZ) drug.

    Methods

    HA-NP was prepared via wet-chemical-method, characterized by XRD, SEM/EDX, TEM, Fourier-transform infrared spectroscopy (FTIR), as well as the measurement of surface area and pore-size distribution. Four drug delivery formulas were prepared in the form of discs (10 x 2 mm) as follows: F1 (MZ/ HA-NP/PMMA), F2 (HA-NP/ PMMA), F3 (control-PMMA) and F4 (MZ/PMMA). Characterization of all formulas was performed using differential scanning calorimetry (DSC) and FTIR. MZ release rate, antimicrobial properties against three oral pathogens, cytotoxicity (MTT assay) and surface micro-hardness were also assessed. Statistical analysis of data was performed using one-way ANOVA test (P < 0.05).

    Results

    DSC thermograms showed compatibility among MZ, HA-NP and PMMA along with physical stability over 6 months storage period at room temperature. FTIR spectroscopy proved the absence of any possible chemical interaction with MZ. MZ-HA-NP/PMMA formula showed relatively better drug release compared to MZ-PMMA. Both formulas showed statistically significant antimicrobial potentials against two microbial strains. MTT demonstrated reduction in cell cytotoxicity after 96 hours with the least value for HA-NP. Surface micro-hardness revealed non-significant reduction compared with the control PMMA.

    Conclusion

    A novel biocompatible drug nanocarrier (HA-NP) was developed and incorporated in PMMA denture base material as a vehicle to allow prolonged sustained drug release to manage oral infections.

    Keywords: PMMA discs, Nanocarriers, HP-NP, Drug delivery, Cytotoxicity, Surface micro-hardness
  • Parvaneh Bayat, Parvaneh Pakravan *, Mojtaba Salouti, Jafar Ezzati Nazhad Dolatabadi Pages 96-103
    Purpose

    Cancer is an example of the most important growing diseases in human society and scientists are trying to treat it without considerable side effects on patient’s health. Solid lipids are colloidal nanoparticles that were used in drug delivery due to their several advantages.

    Methods

    In this work, surface modified targeted solid lipid nanoparticles (SLNs) were fabricated by nano-homogenizer using tripalmitin glyceride and stearic acid as lipid constituents. The size of nanoparticles and morphological evaluations were surveyed using particle size analyzer, scanning electron microscopy; Fourier transforms infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC).

    Results

    The particle size of 148.5 and appropriate polydispersity index were achieved for lipid nanoparticles with an entrapment efficiency of 86.1%. The FT-IR analysis confirmed the coupling of lysine to the free functional group of SLNs. DSC proved the conjugation of amino acid to the surface of carriers. The in vitro epirubicin (EPI) release test exhibited the further controlled release phenomenon for the lysine conjugated nanoparticles. The cytotoxicity assay showed lower IC50 of lysine conjugated SLNs of EPI on the investigated cell line.

    Conclusion

    These studies showed that the fabricated targeted carrier has a very remarkable anticancer effect on breast cancer cell lines in comparison with pure drug.

    Keywords: Solid lipid nanoparticles, Epirubicin, Targeted drug delivery, Cytotoxicity
  • Sahar Yazdani Ashtiani, Saman Ahmad Nasrollahi*, Atefeh Naeimifar, Aubid Nassiri Kashani, Aniseh Samadi, Somayeh Yadangi, Ehsan Aboutaleb, Poorya Abdolmaleki, Rassoul Dinarvand, Alireza Firooz Pages 104-110
    Purpose

    Vitiligo is a long-term common autoimmune disease in which growing patches of skin lose their color. There is no FDA-approved treatment for vitiligo. However, recent studies have demonstrated an immunosuppressive effect on vitiligo lesions in mouse models by simvastatin. A topical formulation was prepared containing simvastatin-loaded nano lipid carriers (simNLCs) for vitiligo treatment followed by evaluating their physicochemical characteristics and clinical safety.

    Methods

    Both the lipid phase and the aqueous phase were heated to 75°C separately, and then simvastatin was dispersed in the lipid phase added to the aqueous phase. The mixture was homogenized for 1 minute, then for Nanostructured Lipid Carriers (NLC) formation, the emulsion was sonicated using a probe sonicator. The simNLCs produced were evaluated for drug entrapment, particle size and morphology, zeta potential, polydispersity index, viscosity, drug content, in vitro drug release, in vivo skin safety test, and long-term stability studies.

    Results

    Dynamic light scattering, transmission electron microscopy and differential scanning calorimetry techniques proved the formation of a stable formulation containing spherical particles with nanoscale size. The drug entrapment efficiency and the drug-loading capacity were determined to be 99.27% and 3.9%, respectively. Human safety results indicated that adding simvastatin to lipid nanoparticles did not cause any changes to skin biophysical parameters.

    Conclusion

    The preparation method of simNLC developed in this study is a suitable method, and the nanoparticles fabricated were safe with acceptable long-term stability and drug entrapment.

    Keywords: Nano lipid carriers, Probe sonication, Simvastatin, Safety, Vitiligo
  • Amrutha Balliamada Monnappa, Jamballi Gangadharappa Manjunatha*, Aarti Sripathi Bhatt *, Kodi Malini Pages 111-119
    Purpose

    Isatin (IS) is a synthetically significant heterocyclic moiety with an influential pharmacodynamic indole nucleus and hence the electrocatalytic property of has been investigated.

    Methods

    The electrochemical analysis was demonstrated by cyclic voltammetry (CV) in the potential window of 0.2 V to 1.4 V using sodium dodecyl sulfate (SDS) modified carbon nano tube paste electrode (SDSMCNTPE) over a pH range of 6 to 8.5 in 0.2 M phosphate buffer solution (PBS). Surface morphology was studied by using Field emission-scanning electron microscopy (FESEM).

    Results

    The CV study discloses that under ideal condition oxidation of IS arises at a potential of 0.970 V accompanied with an exceptional stability, selectivity and sensitivity for the resultant SDSMCNTPE contrasting to bare carbon nano tube paste electrode (BCNTPE). Individual parameters like electrode surface area, effect of surfactant, detection limit, simultaneous detection of IS and resorcinol (RC) were studied at a scan rate of 0.1 V/s. Scan rate study uncovers the process is diffusion controlled. The oxidation peak current amplified linearly with the surge in concentration of IS under ideal condition. Detection limit (LOD) and limit of quantification (LOQ) in the solution of optimum pH (7.5) at a scan rate of 0.100V/s is 2.4×10-7 M and 8.2 × 10-7 M respectively.

    Conclusion

    The proposed electrode portrays excellent repeatability, reproducibility and reliability to resistant electrode fouling.

    Keywords: Cyclic Voltammetry, Carbon nanotube paste electrode, SDS, Isatin, Resorcinol
  • Yandi Syukri *, Annisa Fitria, Suci Hanifah, Muthiah Idrati Pages 120-129
    Purpose

    This study aimed to prepare, characterize, examine the stability and evaluation of the antibacterial activity of Indonesian propolis extract-loaded self-emulsifying (PESE).

    Methods

    Oil, emulsifier, and co-emulsifier were selected as the carrier for the PESE formulation through a propolis-extract solubility test on each carrier, followed by evaluation of the nanoemulsion region in a pseudo ternary phase diagram. Pre-concentrate of PESE was prepared with the addition of 150 mg/mL propolis extract followed by characterization for the transmittance, globule size, zeta potential, thermodynamic stability, robustness to dilution, and accelerated stability. The selected formulation was tested for antibacterial activity using a microdilution method.

    Results

    The PESE characterization produced a clear nanoemulsion with a globule size ranging from 13 to 45 nm and zeta potential of less than −38 mV. The PESE formulation with a composition of 150 mg/mL propolis extract, 20% castor oil, 40%–70% Kolliphor EL, and 10%–40% polyethylene glycol (PEG) 400 were thermodynamically stable. The PESE formulation with the composition of 20% castor oil, 40% Kolliphor EL, and 40% PEG 400 was the optimum formulation that passed the robustness to dilution evaluation and an accelerated stability test for 3 months. The antibacterial activity test on this formulation indicated improved activity against Escherichia coli and Staphylococcus aureus compared with that of propolis extract.

    Conclusion

    These studies demonstrated that PESE in optimum formulation could be used as an antibacterial, particularly in E. coli and S. aureus.

    Keywords: Antibacterial, Castor oil, Propolis extract, Self-emulsifying
  • Débora Antunes Neto Moreno, Mirela Santos Saladini, Fabia Judice Marquesudice Viroel, Murilo Melo Juste Dini, Thaisa Borim Pickler, Jorge Amaral Filho, Carolina Alves dos Santos, Valquíria Miwa Hanai-Yoshida, Denise Grotto, Marli Gerenutti, Stephen Hyslop, Yoko Oshima-Franco * Pages 130-136
    Purpose

    In this work, the potential usefulness of silver nanoparticles (AgNPs) for treating burn wounds was examined.

    Methods

    Second-degree burns were induced in male Wistar rats by touching the skin with a heated (70°C) metallic device for 10 s, after which the animals were randomly allocated to one of two groups: control (n=8, treated with sterile saline) and experimental (n=8, treated with AgNPs, 0.081 mg/mL; 50 µL applied to the burn surface). Seven, 14, 21 and 28 days after lesion induction two rats from each group were killed and blood samples were collected for a complete blood count and to assess oxidative stress. The livers were examined macroscopically and skin samples were collected for histological analysis.

    Results

    Macroscopically, wound healing and skin remodeling in the experimental group were similar to the saline-treated rats. Likewise, there were no significant differences in the histological parameters between the two groups. However, treatment with AgNPs caused a persistent reduction in white blood cell (WBC) counts throughout the experiment, whereas platelet counts increased on days 7 and 28 but decreased on days 14 and 21; there was also an increase in the blood concentration of reduced glutathione on day 7 followed by a decrease on days 21 and 28. There were no significant changes in blood glutathione peroxidase (GSH-Px) and catalase (CAT) activities or in the serum concentration of thiobarbituric acid reactive substances.

    Conclusion

    The findings of this study raise questions about the potential transitory effects of AgNPs based on the changes in WBC and platelet counts, blood glutathione concentrations and macroscopic hepatic alterations.

    Keywords: Hematological parameters, Oxidative stress, Second-degree burn, Silver nanoparticles, Wound healing
  • Sony Valsalan Soba, Merin Babu, Rajitha Panonnummal* Pages 137-149
    Purpose

    Purpose was to improve the skin compatibility and permeability of alpha phellandrene through an ethosomal gel formulation for the treatment of gout; as the oral use of the drug is reported to cause gastrointestinal disturbances and toxicities.

    Methods

    Alpha phellandrene loaded ethosomal formulation (APES) was prepared by cold method for the treatment of gout. APES were loaded into carbopol gel (APEG) by dispersion method. Physico-chemical characterizations of the APES were done by dynamic light scattering (DLS), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR) etc. In vitro release, permeation, haemo-compatibility and anti-inflammatory studies were conducted.

    Results

    APES showed a particle size of 364.83 ± 45.84 nm. The entrapment efficiency of the optimized formulation is found as 95.06 ± 2.51%. Hemolysis data indicated that APES does not cause any significant hemolysis. In vitro drug release studies were carried out using dialysis membrane technique and the amount of drug released from APES & APEG is found to be 95% and 94.21% respectively after 5 and6 hours. Kinetic data analysis revealed that APES & APEG follows first order and zero order release kinetics, respectively. The anti-inflammatory activity studies of the formulation are done by estimating its inhibitory effects on cyclooxygenase II (COX) II, lipoxygenase-5 (LOX-5), Myeloperoxidase (MPO), Inducible nitric oxide synthase (INOS) & cellular nitrite level using RAW 264.7 cells. The significant inhibition in the activities of the enzymes implies the anti-inflammatory activity of the formulations. Skin permeation study was carried out using porcine skin and revealed that the permeation of alpha phellandrene is increased from APES & APEG when compared with alpha-phellandrene solution (APS). Skin deposition study of APS, APES & APEG revealed better drug deposition from APEG (48.799 ± 1.547µg/cm2 ) after 24 hours when compared with APS & APES.

    Conclusion

    Overall results indicate that the ethosomal formulation of alpha phellandrene through transdermal route is an effective alternative for oral use of the drug.

    Keywords: Alpha phellandrene, Terpene, Ethosomal gel, Anti-inflammatory, Gout
  • Sanjeevani Shekhar Deshkar*, Monali Shivaji Jadhav, Satish Vasudeo Shirolkar Pages 150-162
    Purpose

    The present research work was designed to formulate and evaluate carbamazepine (CBZ) loaded nanostructured lipid carrier (NLC) based in-situ gel for nasal delivery.

    Methods

    The NLC formulation of CBZ was prepared by microemulsion technique followed by probe sonication and evaluated for particle size, zeta potential, entrapment efficiency and in vitro drug diffusion. NLC formulation was incorporated into in-situ gelling formulation using poloxamer 407 (P407), poloxamer 188 (P188) and mucoadhesive polymer. The effect of concentration of poloxamer 188 (X1 ), type of mucoadhesive polymer (X2 ) and concentration of mucoadhesive polymer (X3 ) on gelling temperature (Y1 ) and drug diffusion after 8 h (Y2 ) was studied using Box-Behnken design. In vivo anticonvulsant activity of optimized formulation was studied in Wistar rats by maximal electro-convulsion model (MES).

    Results

    The optimized CBZ NLC formulation, with 20% drug loading, 0.5:1 as Precirol:Capmul MCM ratio as lipid phase and 1:3 as Lipid:Smix ratio, resulted in 89.73±0.2% drug entrapment, 55.95±1.09% of drug diffusion after 8 h, particle size of 132.8 nm with polydispersity index of 0.302 and zeta potential of -29.2±6.1 mV. The in-situ gel formulation with 20% P407, 5% P188 and 0.2% chitosan was optimized and demonstrated excellent gelling ability, gelling temperature in the range of 30 to 35°C, 42.46% of drug diffusion in 8 h by Fickian diffusion mechanism and 31.34±0.76% of drug permeation through sheep nasal mucosa. In vitro anticonvulsant activity in MES model in rat demonstrated significant efficacy (71.95% protection against seizure in extension phase) as compared to plain in-situ nasal gel (50.26% protection against seizure in extension phase).

    Conclusion

    NLC based in-situ gelling formulation demonstrated its potential for nasal delivery of CBZ with improved anticonvulsant activity.

    Keywords: Carbamazepine, Nanostructured lipid carrier, In-situ gel, Nasal permeation, Flux, Box-Behnken design
  • Shirin Saberianpour, Reza Rahbarghazi *, Mahdi Ahmadi, Mohammad Nouri, Morteza heidarzadeh, Abbas Karimi, Souror Nemati Pages 163-170
    Purpose

    Here, we investigated the angiogenic potential of endothelial progenitor cells juxtaposed with mesenchymal stem cells (MSCs) inside alginate-gelatin microspheres with stromal derived factor-1α (SDF-1 α) for 7 days.

    Methods

    Six encapsulated groups were allocated including endothelial progenitor cells (EPCs), EPCs/SDF-1α, MSCs, MSCs/SDF-1α, EPCs+MSCs and EPCs+MSCs/SDF-1α. Cells were encapsulated with a mixture of 1% alginate and 2% gelatin hydrogel. Cell survival was examined by MTT assay. Endothelial differentiation was determined by flow cytometry and ELISA. Tubulogenesis assay and Ac-Dil-LDL uptake were used to detect functional activity. Cell migration was analyzed by Transwell insert and gelatin zymography analyses. By using real-time polymerase chain reaction (PCR), we measured the transcription of Akt and PK1.

    Results

    We found an increase in cell viability in MSCs/SDF-1α microspheres compared to EPCs group (P<0.05). EPC/MSCs co-culture contributed to the increase of CD133+ cells while we found high CD31 levels in MSCs group (P<0.05). Juxtaposition of EPC with MSCs increased tubulogenesis compared to SDF-1a-free condition (P<0.001). SDF-1α had the potential to increase in AC-LDL uptake in MSCs and EPCs/MSCs groups. Cells migration and MMP-9 activities increased after treatment with SDF-1α. SDF-1α upregulated PK1 and Akt in encapsulated cells, especially in a co-culture system.

    Conclusion

    Alginate-gelatin microspheres could alter the angiogenic potential of progenitor cells in the presence of SDF-1α

    Keywords: Endothelial Progenitor Cells, Mesenchymal Stem Cells, Alginate-gelatin microspheres, Angiogenesis
  • Yandi Syukri*, Muhammad Taher, Ronny Martien, Endang Lukitaningsih, Agung Endro Nugroho, ZainulAmiruddin Zakaria Pages 171-180
    Purpose

    Insulin resistance is a characteristic of non-insulin-dependent diabetes mellitus associated with obesity and caused by the failure of pancreatic beta cells to secrete sufficient amount of insulin. Andrographolide (AND) improves beta-cell reconstruction and inhibits fat-cell formation. This research aimed to improve the delivery of water-insoluble AND in self-nanoemulsifying (ASNE) formulation, tested in streptozotocin (STZ)-induced diabetic rats and 3T3-L1 preadipocyte cells.

    Methods

    A conventional formulation of AND in suspension was used as a control. The experimental rats were orally administered with self-nanoemulsifying (SNE) and suspension of AND for 8 days. Measurements were performed to evaluate blood glucose levels in preprandial and postprandial conditions. Immunohistochemistry was used to assess the process of islet beta cell reconstruction. In vitro study was performed using cell viability and adipocyte differentiation assay to determine the delivery of AND in the formulation.

    Results

    ASNE lowered blood glucose levels (day 4) faster than AND suspension (day 6). The histological testing showed that ASNE could regenerate pancreatic beta cells. Therefore, ASNE ameliorated pancreatic beta cells. The in vitro evaluation indicated the inhibition of adipocyte differentiation by both AND and ASNE, which occurred in a time-dependent manner. ASNE formulation had better delivery than AND.

    Conclusion

    ASNE could improve the antidiabetic activity by lowering blood glucose levels, enhancing pancreatic beta cells, and inhibiting lipid formation in adipocyte cells.

    Keywords: Adipocyte differentiation, Andrographolide, Self-nanoemulsifying, Pancreatic beta cells
  • Elfahmi Elfahmi*, Rizqiya Astri Hapsari, Tamara Chrysanthy, Vaniarta Synthiarini, Fifi Fitriyah Masduki, Agus Setiawan, Toshiya Muranaka Pages 181-187
    Purpose

    Artemisinin, a secondary metabolite in Artemisia annua is one of primary choice for the treatment of malaria, it is naturally produced in low concentration from this plant. This study was aimed to clone key enzymes of artemisinin production in order to enhance its production through the semi-synthetically production in Saccharomyces cerevisiae.

    Methods

    Two key enzymes in artemisinin biosynthetic pathway which are farnesyl phosphate synthase (fps) and amorpha-4,11-diene synthase (ads) genes were transformed into S. cerevisiae using pBEVY vector. Successful transformation was checked by polymerase chain reaction (PCR) method and sequencing analysis

    Results

    Recombinant plasmids which are pBEVY-GU_ads and pBEVY_GL_fps were successfully constructed. The optimized ads gene was amplified using PCR with a couple of primers that are designed in order to provide the homolog recombination between ads gene with the expression plasmid of pBEVY-GU respectively. While the A. annua optimized fps gene was cloned using classical method. Transformants were grown in selective media Synthetic Defined (SD) without leucine for transformants contain plasmid pBEVY-GL_fps and media without uracil for transformants contain plasmid pBEVY-GU_ads. Confirmation of colonies was done by PCR with primers to amplify fps and ads. DNA from yeast was isolated from positive colonies then transformed to E. coli. Plasmid from E. coli was isolated for restriction analysis and sequencing. Protein expression was induced by cultivating the yeast in the media with 2% galactose.

    Conclusion

    Based on PCR, restriction and sequencing analysis, it could be concluded that fps and ads genes were successfully constructed, transformed and expressed in S. cerevisiae.

    Keywords: Amorpha-4, 11-diene synthase, Artemisinin, Farnesyl phosphate synthase, Malaria, Saccharomyces cerevisiae
  • Edy Meiyanto*, Ulfatul Husnaa, Ria Fajarwati Kastian, Herwandhani Putri, Yonika Arum Larasati, Annisa Khumaira, Dyaningtyas Dewi Putri Pamungkas, Riris Istighfari Jenie, Masashi Kawaichi, Beni Lestari, Takashi Yokoyama, Jun-ya Kato Pages 188-196
    Purpose

    The current study aims to evaluate the in vitro cytotoxic and cell migration effects of synthetic curcumin and its analogues on HER2 and nuclear factor kappa B (NFκB) pathways, as well as the in vivo inhibitory effect on cancer growth of metastatic breast cancer.

    Methods

    Cell viability, protein expression, and protein localization were determined in vitro using MTT assay, western blotting, and immunofluorescence, respectively. Meanwhile, scratch wound healing assay and gelatin zymography were conducted to investigate the metastasis inhibitory effect. The in vivo anti-tumor ability was evaluated in xenograft mouse model using triple-negative breast cancer (TNBC) cells.

    Results

    Curcumin, PGV-0, and PGV-1 exhibited cytotoxic effect against HER2-overexpressing breast cancer cells. Although PGV-1 showed the best activity in the single cytotoxic assay, curcumin showed the strongest synergism with doxorubicin. Curcumin and PGV-0 inhibited membrane localization of HER2. In contrast, PGV-1 neither inhibited localization nor decreased the expression of HER2, nonetheless showed the most potent inhibition against nuclear localization of p65 indicating the different mechanisms of curcumin, PGV-0, and PGV-1. Regarding cancer metastasis, curcumin and PGV-1 showed inhibitory activities against cell migration and inhibited MMP-2 and MMP-9 protein expression. Lastly, PGV-1 was more potent compared to curcumin to suppress the tumor formation of metastatic breast cancer xenograft model in nude mice.

    Conclusion

    Overall, our study strengthens the potency of curcumin analogue, PGV-1, for treating several types of cancer, including metastatic breast cancer.

    Keywords: Breast cancer, Curcumin analogues, HER2, Metastasis, NFκB, Triple-negative breast cancer
  • Hossein Mehralizadeh, Mohammad Reza Aliparasti, Mehdi Talebi, Shabnam Salekzamani, Shohreh Almasi, Morteza Raeisi, Mehdi Yousefi, AliAkbar Movassaghpour* Pages 197-203
    Purpose

    Acute myeloid leukemia (AML) is the most prevalent acute leukemia in adults. It possesses different cytogenetic and molecular features. The expression of Wilms tumor-1 (WT1), brain and acute leukemia, cytoplasmic (BAALC) and ETS-related gene (ERG) might be considered as prognostic factors in AML patients. The aim of this study was to determine the mRNA expressions of WT-1, BAALC and ERG genes in bone marrow of mononuclear cells and their effects on complete remission in the Iranian AML patients, pre- and post- chemotherapy.

    Methods

    Forty AML patients with normal karyotype were evaluated. The mRNA gene expressions were measured with quantitative real-time PCR in bone marrow of mononuclear cells of AML patients at the baseline and after chemotherapy. The subtypes of AML and flow cytometry panel were also assessed. Complete remission (CR) after the treatment was addressed for all patients.

    Results

    The mRNA expressions of WT-1, BAALC and ERG were significantly decreased after the treatment (p= 0.001, 0.017, 0.036). WT-1 mRNA expression was inversely correlated with CR after chemotherapy (P =0.024). There was also significant correlation between baseline expression of BAALC and CR (P=0.046). No significant correlation was observed between ERG and CR pre- and post- chemotherapy (P =0.464 and 0.781). There was also significant correlation between BAALC mRNA expression and CD34+ (P <0.001).

    Conclusion

    The present study showed that WT-1 decreased significantly after standard chemotherapy which could have favorable effects on CR. Also, the high expression of BAALC could have a poor prognostic role in AML patients. The identification of these gene expressions can be an efficient approach in targeted therapy among AML patients.

    Keywords: Acute myeloid leukemia, WT-1, BAALC, ERG, Gene expression, Chemotherapy
  • Solmaz Ghaffari*, Faezeh Alihosseini, Seyed Mahdi Rezayat Sorkhabadi, Sepideh Arbabi Bidgoli, Seyyedeh Elaheh Mousavi, Setareh Haghighat, Ahoo Afshar Nasab, Nooshin Kianvash Page 204
  • Sara Shojaei Zarghani, Samin Abbaszadeh, Mohammad Alizadeh, Maryam Rameshrad, Alireza Garjani, Hamid Soraya* Page 205