Overview of Physicochemical Properties of Nanoparticles as Drug Carriers for Targeted Cancer Therapy.

The advent of nanotechnology has brought about revolutionary innovations in biological research techniques and medical practice. In recent years, various "smart" nanocarriers have been introduced to deliver therapeutic agents specifically to the tumor tissue in a controlled manner, thereby minimizing their side effects and reducing both dosage and dosage frequency. A large number of nanoparticles have demonstrated initial success in preclinical evaluation but modest therapeutic benefits in the clinical setting, partly due to insufficient delivery to the tumor site and penetration in tumor tissue. Therefore, a precise understanding of the relationships betweenthe physicochemical properties of nanoparticles and their interaction with the surrounding microenvironment in the body is extremely important for achieving higher concentrations and better functionality in tumor tissues. This knowledge would help to effectively combine multiple advantageous functions in one nanoparticle. The main focus of the discussion in this review, therefore, will relate to the main physicochemical properties of nanoparticles while interacting within the body and their tuning potential for increased performance.

Wharton Jelly Derived Mesenchymal Stem Cell's Exosomes Demonstrate Significant Antileishmanial and Wound Healing Effects in Combination with Aloe-Emodin: An in Vitro Study.

The aim of the present study was to explore the antileishmanial performance and wound healing effect of exosomes isolated from Wharton Jelly derived mesenchymal stem cells (WJ-MSCs) in combination with aloe-emodin. MSCs obtained from Wharton Jelly were characterized by flow cytometry. Exosomes were isolated from cultivated stem cells by ultacentrifugation method. Scanning Electron Microscopy (SEM), Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA) and flow cytometry were used for characterization of obtained exosomes. The cytotoxicities of characterized exosomes and aloe-emodin at different concentrations were investigated on L929 and J774 cell lines. Non-toxic concentrations of each agent were combined and their inhibitory efficacies on L.major promastigotes and amastigotes were investigated by different techniques such as MTT, parasite count and measurements of infection index. Finally, wound healing activities of combinations were examined on in vitro artifical wound model and compared with the use of exosomes alone. According to outcome of flow cytometic analysis, vesicles isolated from WJ-MSCs highly expressed the markers such as CD63 special for exosome profile. SEM and NTA results demonstrated that derived exosomes possessed dimensions between 150 to 200 nanometers and elicited the cup-shape specific to exosomes. Combinations including non-toxic dosages of exosomes and aloe-emodin demonstrated superior antileishmanial effectivenesses both on promastigotes and amastigotes in contrast to use of exosome alone since they lead to inhibition of promastigotes and amastigotes for 4 and 10-folds in comparison to control, respectively. Additionally, combinations elicited more rapidly and effective in vitro wound-healing performance in contrast to use of exosome alone. At the end of 24 h incubation application of combinations gave rise to wound closure at a rate of 72 %, while in the control group 52 % of wound area has not been healed, yet. These results reflect that mentioned combination has great potential to be used in treatment of cutaneus leishmaniasis (CL) since they have magnificient capacity to inhibit Leishmania parasites while enhancing wound healing.

Formulating and Characterizing an Exosome-based Dopamine Carrier System.

Exosomes between 40 and 200 nm in size constitute the smallest subgroup of extracellular vesicles. These bioactive vesicles secreted by cells play an active role in intercellular cargo and communication. Exosomes are mostly found in body fluids such as plasma, cerebrospinal fluid, urine, saliva, amniotic fluid, colostrum, breast milk, joint fluid, semen, and pleural acid. Considering the size of exosomes, it is thought that they may play an important role in central nervous system diseases because they can pass through the blood-brain barrier (BBB). Hence, this study aimed to develop an exosome-based nanocarrier system by encapsulating dopamine into exosomes isolated from Wharton's jelly mesenchymal stem cells (WJ-MSCs). Exosomes that passed the characterization process were incubated with dopamine. The dopamine-loaded exosomes were recharacterized at the end of incubation. Dopamine-loaded exosomes were investigated in drug release and cytotoxicity assays. The results showed that dopamine could be successfully encapsulated within the exosomes and that the dopamine-loaded exosomes did not affect fibroblast viability.

A nanotechnology-based new approach in the treatment of breast cancer: Biosynthesized silver nanoparticles using Cuminum cyminum L. seed extract.

The present study reports the anticancer activities of Cuminum cyminum L. (Cumin) seed extract, chemically synthetized silver nanoparticles (AgNPs) and biosynthesized silver nanoparticles (Bio-AgNPs) from Cumin seeds on human breast adenocarcinoma cell line (MCF-7) and human breast adenocarcinoma metastatic cell line (AU565). The synthetized nanoparticles were characterized by dynamic light scattering (DLS), UV-visible spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). The cytotoxic and anticancer effects of AgNPs and Bio-AgNPs were determined by MTT assay. According to the cytotoxicity analysis, Bio-AgNPs appears to be less toxic against J774 macrophage cells than AgNPs since IC50 values were measured as 0.75 and 1.25 µg/ml for AgNPs and Bio-AgNPs, respectively. On the other hand, Bio-AgNPs demonstrated significant inhibitory effects on human breast cancer cells at non-toxic concentrations such as 0.25 and 0.5 µg/ml. However, at increased concentrations, the lethal effects of AgNPs on breast cancer cells were higher than Bio-AgNPs. When cytotoxic and anticancer characteristics of Cumin extract were investigated, it was established that it did not show any inhibitory effect on J774 cells, while killing the half of MCF-7 cells at investigated concentrations. Interestingly, Cumin extract gave rise to no inhibitory effects against AU565 cells. On the other hand, AgNPs and Bio-AgNPs exhibited considerable anticancer activities on both cell lines. The inhibition percentages of AgNPs on MCF-7 and AU565 cell lines were respectively evaluated as 95% and 97% at the highest concentrations applied (12.5 µg/ml). Similarly, we determined that 87.5% and 96% of MCF-7 and AU565 cells were respectively inhibited when they were exposed to the highest concentrations of Bio-AgNPs. Considering relatively toxic-free features of Bio-AgNPs prepared from Cuminum cyminum L. seed extracts, it can be thought that this formulation will be a pioneer in development of nanotechnology-based new anticancer drug for the treatment of breast cancer in near future.

Current Approaches in Treatment of Diabetic Retinopathy and Future Perspectives.

Diabetes mellitus (DM) is a metabolic disease, which is the most common cause of low vision in developing countries and affects almost all systems of the body. In view of the increase in DM prevalence in the world, it would not be a surprise that diabetic retinopathy (DR) and other vascular complications related to diabetes become a serious public health problem. Currently, vascular endothelial growth factor, laser photocoagulation, and intravitreal steroids are the mainstays for DR treatment, but the efficacy of these treatment strategies remains insufficient. Therefore, new treatment modalities for DR have been developed, such as stem cell therapies, extracellular vesicular system, and nanodrug delivery systems. Although there have been several reviews in the literature on the treatment of DR, we have not confronted any review that has the titles of all these topics. With this review, we aim to present the pathophysiology of DR and to review the current and promising new treatment methods based on stem cells, extracellular vesicular system, and nanodrug delivery systems for the future of DR management.

An overview of nanotechnology-based treatment approaches against Helicobacter Pylori.

Introduction: Helicobacter Pylori (H.Pylori) is a pathogen that infects about 50% of the world's population and is known to be responsible for gastroduodenal diseases such as atrophic gastritis, peptic ulcer and stomach cancer. Nowadays, there is no treatment that ensures complete eradication. In addition, resistance to antibiotics used in the current treatment adversely affects the success rates in the fight against infection. Areas covered: This article take attention to treatment approaches using nanoparticles as an alternative to H.Pylori treatment to cope with increased antibiotic resistance. The purpose of this review is to provide an overview of the current limitations and new promising altenatives in treatment of H.Pylori, to highlight the location of nanotechnology to overcome treatment failures, and to emphasize the advantages of using membrane-coated nanoparticles for the first time. Expert opinion: Because of the current problems in the treatment of H.Pylori, there is increasing interest in alternative approaches including nanotechnology. The strong antibacterial effects of metallic nanoparticles, the advantages of polymeric nanoparticles in drug delivery and drug protection, and the prominent properties of membrane-coated nanoparticles in direct targeting demonstrate the significance of nanotechnology in developing new approaches for treatment of H.Pylori infection.

Current aspects in treatment of breast cancer based of nanodrug delivery systems and future prospects.

Breast cancer is one of the most common diseases worldwide. The risk of getting this disease in female is 30% and the mortality rate is 14%. The breast cancer treatment is based on surgery, chemotherapy and radiotherapy. However, an effective treatment method has not been developed. The main cause of failure in the treatment is cancer stem cells metastasis and chemo-resistance. The use of nanocarrier systems against breast cancer stem cells has great importance. Not only advantages of polymeric drug delivery systems are increasing the stability and reduce the side effects of drugs, but also they have disadvantages such as biocompatibility and long-term potential safety. However, in recent years, studies on exosomes provide several advantages. Exosomes usage as nanocarrier do not cause immunological reactions also the drug effectively transport into the cytosol of targeted cell and have more stability characteristics. Although there are studies about various nanocarrier systems in literature against breast cancer but in general, we have not found any review that brings them together and develops a systematic approach to solving the problem. This review mentions prospective new strategies based on various nanocarrier systems and emphasize the importance of exosome based on drug delivery systems in the treatment of breast cancer.

Problems in Stem Cell Therapy for Cardiac Repair and Tissue Engineering Approaches Based on Graphene and Its Derivatives.

BACKGROUND: Today, coronary artery disease is still one of the most important causes of mortality despite advanced surgical methods, pharmacotherapies and organ transplantation. These treatment modalities are intended to prevent further progression of myocardial infarction and do not involve the repair of the damaged part. Therefore, stem cell therapy has emerged as a new approach for the treatment of coronary artery disease. However, there are some restrictions that limit the use of these cells for desired repair. The leading limitation is that newly formed cardiomyocytes do not provide electrical integrity with local cells. OBJECTIVE: In this paper, we review the difficulties that limit the use of stem cell therapy in cardiac repair and emphasize the importance of the integration of stem cell with tissue scaffolds with conductivity. Furthermore, significance of using graphene scaffolds in cardiac tissue engineering is highlighted due to its conductivity features. RESULT: Recently, the fabrication of tissue scaffoldings has made it possible to create a biomimetic cellular environment while providing a new approach to solving these problems in treatment. Especially, the integration of stem cell therapy with graphene-based tissue scaffolds with electrical conductivity, is one of the promising new strategies to turn the success of two approaches of tissue engineering into synergistic effect in cardiac repair. CONCLUSION: Literature analysis has demonstrated that there are some limitations in use of stem cell therapy for successful treatment of cardiac repair and graphene-based tissue engineering approaches which are promising to solve these problems in the near future.

Current Approaches in Development of Immunotherapeutic Vaccines for Breast Cancer.

Cancer is the leading cause of death worldwide. In developed as well as developing countries, breast cancer is the most common cancer found among women. Currently, treatment of breast cancer consists mainly of surgery, chemotherapy, hormone therapy, and radiotherapy. In recent years, because of increased understanding of the therapeutic potential of immunotherapy in cancer prevention, cancer vaccines have gained importance. Here, we review various immunotherapeutic breast cancer vaccines including peptide-based vaccines, whole tumor cell vaccines, gene-based vaccines, and dendritic cell vaccines. We also discuss novel nanotechnology-based approaches to improving breast cancer vaccine efficiency.

Nigella sativa oil entrapped polycaprolactone nanoparticles for leishmaniasis treatment.

This study is the first to investigate the antileishmanial activities of Nigella sativa oil (NSO) entrapped poly-ɛ-caprolactone (PCL) nanoparticles on Leishmania infantum promastigotes and amastigotes in vitro. NSO molecules with variable initial doses of 50, 100, 150, and 200 mg were successfully encapsulated into PCL nanoparticles identified as formulations NSO1, NSO2, NSO3, and NSO4, respectively. This process was characterised by scanning electron microscope, dynamic light scattering, Fourier transform infrared, encapsulation efficiency measurements, and release profile evaluations. The resulting synthetised nanoparticles had sizes ranging between 200 and 390 nm. PCL nanoparticles encapsulated 98% to 80% of initial doses of NSO and after incubation released approximately 85% of entrapped oil molecules after 288 h. All investigated formulations demonstrated strong antileishmanial effects on L. infantum promastigotes by inhibiting up to 90% of parasites after 192 h. The tested formulations decreased infection indexes of macrophages in a range between 2.4- and 4.1-fold in contrast to control, thus indicating the strong anti-amastigote activities of NSO encapsulated PCL nanoparticles. Furthermore, NSO-loaded PCL nanoparticles showed immunomodulatory effects by increasing produced nitric oxide amounts within macrophages by 2-3.5-fold in contrast to use of free oil. The obtained data showed significant antileishmanial effects of NSO encapsulated PCL nanoparticles on L. infantum promastigotes and amastigotes.

Meglumine antimoniate-TiO2@Ag nanoparticle combinations reduce toxicity of the drug while enhancing its antileishmanial effect.

Currently, the treatment of leishmaniasis is increasingly insufficient as current antileishmanial drugs have many disadvantages such as toxic side effects, high cost, and growing drug resistance. In order to overcome these disadvantages, researchers have recently focused on combination therapy by using pentavalent antimonials in conjunction with other antileihmanial compounds. Our previous study found that TiO2@Ag nanoparticles (TiAgNps) demonstrated significant antileishmanial effects. However, a lethal dose of TiAgNps on L. topica promastigotes was found to be toxic for macrophage cells. Moreover, non-toxic concentrations of TiAgNps were ineffective in inhibiting L. topica promastigotes and amastigotes. Thus, we propose the use of TiAgNps in combination with other antileishmanial compounds like meglumine antimoniate (MA) at non-toxic concentrations, which may increase the efficacies of both agents and decrease their toxicities. Therefore, the aim of this study was to determine in vitro and in vivo antileishmanial efficacies of TiAgNps-MA combinations at non-toxic concentrations and develop a new approach for treatment that lowers the toxicities of pentavalent antimonials to minimal levels and enhances their effectiveness. In vitro screening was performed on L. topica promastigote and amastigote-macropage culture by using MTT assay to determine proliferation, perform infection index analysis, and to conduct a Griess reaction for nitric oxide production, while in vivo antileishmanial assays were applied on Balb/c mice with CL models. The results demonstrated that combinations including TiAgNps and MA at non-toxic concentrations were highly efficacious against both promastigotes and amastigotes, while MA application alone did not show any inhibitory effects. It was determined that combination applications decreased the proliferation of L. topica promastigotes 2- to 5-fold in contrast to use of MA alone, and was dependent on concentrations. Moreover, the use of combinations led to inhibition of L. topica amastigotes at rates ranging between 80% and 95%. Additionally, combinations were found to decrease metabolic activities of each form of the parasite at ranges between 7- to 20-fold, causing programmed-cell death and stimulation of macrophages for intensive production of nitric oxide, which is accepted as an important antileishmanial agent (p<0.05). Furthermore, Σ FIC analysis demonstrated that the tested combinations composed little additive, but mostly synergistic effects for inhibition of promastigotes and amastigotes. According to in vivo screening results, the combinations displayed high antileishmanial activities by successfully healing lesions and significantly reducing parasite burdens. Combined, these results show that TiAgNps-MA combinations were much more effective than use of MA alone at non-toxic concentrations and they possess high potential for development of new antileishmanial drugs to fight against leishmaniasis.

A nanotechnology based new approach for chemotherapy of Cutaneous Leishmaniasis: TIO2@AG nanoparticles - Nigella sativa oil combinations.

Since toxicity and resistance are the major drawbacks of current antileishmanial drugs, studies have been recently focused on combination therapy in fight against leishmaniasis. Combination therapy generally provides opportunity to decrease toxicity of applied agents and enhance their antimicrobial performance. Moreover, this method can be effective in preventing drug resistance. Highly antileishmanial effects of silver doped titanium dioxide nanoparticles (TiAgNps) and Nigella sativa oil were demonstrated in previous studies. However, toxicity is still an important factor preventing use of these molecules in clinic. By considering high antileishmanial potential of each agent and basic principles of combination therapy, we propose that use of combinations including non-toxic concentrations of TiAgNps and N. sativa oil may compose more effective and safer formulations against Leishmania parasites. Therefore, the main goal of the present study was to investigate antileishmanial effects of non-toxic concentrations of TiAgNps and Nigella sativa oil combinations on promastigote and amastigote-macrophage culture systems and also to develop nanotechnology based new antileishmanial strategies against Cutaneous Leishmaniasis. Numerous parameters such as proliferation, metabolic activity, apoptosis, amastigote-promastigote conversion, infection index analysis and nitric oxide production were used to detect antileishmanial efficacies of combinations. Investigated all parameters demonstrated that TiAgNps-N. sativa oil combinations had significant antileishmanial effect on each life forms of parasites. Tested combinations were found to decrease proliferation rates of Leishmania tropica promastigotes in a range between 1,5-25 folds and metabolic activity values between 2 and 4 folds indicating that combination applications lead to virtually inhibition of promastigotes and elimination of parasites were directly related to apoptosis manner. TiAgNps-N. sativa combinations also demonstrated killing effects on L. tropica amastigotes by decreasing infection index values of macrophages 5-20 folds, inhibiting their metabolic activities up to 5 fold, preventing amastigote-promastigote conversion and producing high amounts of nitric oxide. All these results emphasize high potential of TiAgNps-N. sativa oil combinations as new, safer and effective antileishmanial formulations against Cutaneous Leishmaniasis.

Human olfactory stem cells for injured facial nerve reconstruction in a rat model.

BACKGROUND: The purpose of this study was to show the efficacy of olfactory stem cells for injured facial nerve reconstruction in a rat model. METHODS: Olfactory stem cells were isolated from the olfactory mucosa of human participants. A 2-mm excision was performed on the right facial nerve of all rats. Reconstruction was performed with a conduit in group 1 (n = 9); a conduit and phosphate-buffered saline in group 2 (n = 9); and a conduit and labeled olfactory stem cell in group 3 (n = 9). Rats were followed for whisker movements and electroneuronography (ENoG) analyses. RESULTS: The whisker-movement scores for group 3 were significantly different from other groups (p < .001). ENoG showed that the amplitude values for group 3 were significantly different from group 1 and group 2 (p = .030; p < .001). Group 3 showed marked olfactory stem cell under a fluorescence microscope. CONCLUSION: This study suggests that olfactory stem cells may be used as a potent cellular therapy for accelerating the regeneration of peripheral nerve injuries. © 2016 Wiley Periodicals, Inc. Head Neck 38: E2011-E2020, 2016.

The effect of CO2 laser beam welded AISI 316L austenitic stainless steel on the viability of fibroblast cells, in vitro.

It has been determined by the literature research that there is no clinical study on the in vivo and in vitro interaction of the cells with the laser beam welded joints of AISI 316L biomaterial. It is used as a prosthesis and implant material and that has adequate mechanical properties and corrosion resistance characteristics. Therefore, the interaction of the CO2 laser beam welded samples and samples of the base metal of AISI 316L austenitic stainless steel with L929 fibroblast cells as an element of connective tissue under in vitro conditions has been studied. To study the effect of the base metal and the laser welded test specimens on the viability of the fibroblast cells that act as an element of connective tissues in the body, they were kept in DMEMF-12 medium for 7, 14, 28 days and 18 months. The viability study was experimentally studied using the MTT method for 7, 14, 28 days. In addition, the direct interaction of the fibroblast cells seeded on 6 different plates with the samples was examined with an inverted microscope. The MTT cell viability experiment was repeated on the cells that were in contact with the samples. The statistical relationship was analyzed using a Tukey test for the variance with the GraphPad statistics software. The data regarding metallic ion release were identified with the ICP-MS method after the laser welded and main material samples were kept in cell culture medium for 18 months. The cell viability of the laser welded sample has been detected to be higher than that of the base metal and the control based on 7th day data. However, the laser welded sample's viability of the fibroblast cells has diminished by time during the test period of 14 and 28 days and base metal shows better viability when compared to the laser welded samples. On the other hand, the base metal and the laser welded sample show better cell viability effect when compared to the control group. According to the ICP-MS results of the main material and laser welded samples which were kept in the cell culture medium for 18 months, it was determined that the Fe, Ni and Cr ion concentration released to the cell culture medium from the laser welded test sample was less than that of the main material.

Preparation, characterization and immunological evaluation: canine parvovirus synthetic peptide loaded PLGA nanoparticles.

BACKGROUND: Canine parvovirus 2 (CPV-2) remains a significant worldwide canine pathogen and the most common cause of viral enteritis in dogs. The 1 L15 and 7 L15 peptides overlap each other with QPDGGQPAV residues (7-15 of VP2 capsid protein of CPV) is shown to produce high immune response. PLGA nanoparticles were demonstrated to have special properties such as; controlled antigen release, protection from degradation, elimination of booster-dose and enhancing the cellular uptake by antigen presenting cells. Nevertheless, there is no study available in literature, about developing vaccine based on PLGA nanoparticles with adjuvant properties against CPV. Thus, the aim of the present study was to synthesize and characterize high immunogenic W-1 L19 peptide (from the VP2 capsid protein of CPV) loaded PLGA nanoparticle and to evaluate their in vitro immunogenic activity. RESULTS: PLGA nanoparticles were produced with 5.26 ± 0.05 % loading capacity and high encapsulation efficiency with 81.2 ± 3.1 %. Additionally, it was evaluated that free NPs and W-1 L19 peptide encapsulated PLGA nanoparticles have Z-ave of 183.9 ± 12.1 nm, 221.7 ± 15.8 nm and polydispersity index of 0.107 ± 0.08, 0.135 ± 0.12 respectively. It was determined that peptide loaded PLGA nanoparticles were successfully phagocytized by macrophage cells and increased NO production at 2-folds (*P < 0.05) in contrast to free peptide, and 3-folds (*P < 0.01) in contrast to control. CONCLUSION: In conclusion, for the first time, W-1 L19 peptide loaded PLGA nanoparticles were successfully synthesized and immunogenic properties evaluated. Obtained results showed that PLGA nanoparticles enhanced the capacity of W-1 L19 peptide to induce nitric oxide production in vitro due to its adjuvant properties. Depend on the obtained results, these nanoparticles can be accepted as potential vaccine candidate against Canine Parvovirus. Studies targeting PLGA nanoparticles based delivery system must be maintained in near future in order to develop new and more effective nano-vaccine formulations.

Isolation and diagnosis of Helicobacter pylori by a new method: microcapillary culture.

AIM: To investigate the performance of the microcapillary culture method (MCM) in Helicobacter pylori (H. pylori) isolation and diagnosis. METHODS: Microcapillary culture (MC), classical culture (CC), rapid urease (CLO) test, and histopathologic examination (HE) were performed with biopsy samples. Homogenized biopsy samples were loaded into capillary tubes and incubated for 48 h at 37 °C without providing a microaerophilic environment. Additionally, three or four loops of the homogenized sample were inoculated in a ready-to-use selective medium (Becton Dickinson, Helicobacter Agar, Modified) specific for the isolation of H. pylori and incubated at 37 °C in a microaerophilic atmosphere provided by CampyGen (Becton Dickinson, GasPack). Bacteria reproducing in microcapillary tubes were evaluated in an inverted microscope and also were evaluated after performing a CC with the content. Results obtained by CC, CLO test, and HE were compared with those of MC. The diagnostic performances of the methods used in this study were evaluated for specificity, sensitivity, positive predictive value (PPV), negative predictive value (NPV), and CI. RESULTS: H. pylori was found positive by CLO test + HE and/or CC culture in 26 patient antrum and corpus biopsy samples. In 25 (25/26) patient biopsy samples, H. pylori was isolated by MCM, whereas in only 14 (14/26) patient biopsy samples, H. pylori was isolated by CC. CLO test and HE were found positive in 17 (17/26) patient biopsy samples. Comparing the results of the isolation of H. pylori by MCM, CC, CLO test, and HE, the sensitivity of the MCM was found as 96%, the specificity as 80%, the PPV as 83%, the NPV as 95%, and the 95%CI as 0.76 (χ (2) = 31.51, P < 0.01) whereas the sensitivity of the CC was found as 54% (χ (2) = 19.15, P < 0.01), and the sensitivity of the CLO test and HE were found as 65% (χ (2) = 25.26, P < 0.01). CONCLUSION: This new microcapillary cultivation method for H. pylori has high diagnostic sensitivity compared with CC, HE, and CLO tests.

Synthesis and characterization of antigenic influenza A M2e protein peptide-poly(acrylic) acid bioconjugate and determination of toxicity in vitro.

The influenza A virus is a critical public health problem that causes epidemics and pandemics, and occurs widely all over the world. Various vaccines against the virus have not provided a solution to the problem. Different approaches, particularly M2e peptide-based vaccines, are available for developing universal vaccines against influenza A. However, it is important to select a suitable carrier to obtain an effective vaccine. Accordingly, studies on the usage of various carriers are ongoing. Particularly, polymer-based carriers have gained importance due to both drug delivery and adjuvant effects. Therefore, bioconjugate of the M2e protein peptide from the influenza A virus covalent bonded with poly(acrylic) acid was synthesized in our study for the first time. The characterization was performed using size-exclusion chromatography and fluorescence spectroscopy; subsequently, it was found that the bioconjugate of the examined lower doses (0.05 and 0.5 mg/ml) have no toxic effects on human cell lines. These results suggest that, in the future, the poly(acrylic) acid bioconjugate of the M2e peptide should be studied in vivo for universal vaccine development against the influenza A virus.

Investigation of antileishmanial activities of Tio2@Ag nanoparticles on biological properties of L. tropica and L. infantum parasites, in vitro.

Leishmaniasis is a public health problem which is caused by protozoon parasites belonging to Leishmania species. The disease threatens approximately 350 million people in 98 countries all over the world. Cutaneous Leishmaniasis (CL) and Visceral Leishmaniasis (VL) are the mostly commonly seen forms of the disease. Treatment of the disease has remained insufficient since current antileishmanial drugs have several disadvantages such as toxicity, costliness and drug-resistance. Therefore, there is an immediate need to search for new antileishmanial compounds. TiO2@Ag nanoparticles (TiAg-Nps) have been demonstrated as promising antimicrobial agents since they provide inhibition of several types of bacteria. The basic antimicrobial mechanism of TiAg-Nps is the generation of reactive oxygen species (ROS). Even though Leishmania parasites are sensitive to ROS, there is no study in literature indicating antileishmanial activities of TiAg-Nps. Herein, in this study, TiAg-Nps are shown to possess antileishmanial effects on Leishmania tropica and Leishmania infantum parasites by inhibiting their biological properties such as viability, metabolic activity, and survival within host cells both in the dark and under visible light. The results indicate that TiAg-Nps decreased viability values of L. tropica, and L. infantum promastigotes 3- and 10-fold, respectively, in the dark, while these rates diminished approximately 20-fold for each species in the presence of visible light, in contrast to control. On the other hand, non-visible light-exposed TiAg-Nps inhibited survival of amastigotes nearly 2- and 2.5-fold; while visible light-exposed TiAg-Nps inhibited 4- and 4.5-fold for L. tropica and L. infantum parasites, respectively. Consequently, it was determined that non-visible light-exposed TiAg-Nps were more effective against L. infantum parasites while visible light-exposed TiAg-Nps exhibited nearly the same antileishmanial effect against both species. Therefore, we think that a combination of TiAg-Nps and visible light can be further used for treatment of CL, while application of TiAg-Nps alone can be a promising alternative in VL treatment.

Generation of avirulent Leishmania parasites and induction of nitric oxide production in macrophages by using polyacrylic acid.

Polyacrylic acid (PAA) is one of the anionic synthetic polyelectrolytes and is used in various immunological and pharmaceutical applications. PAA has been used as adjuvant in veterinary vaccines, in particular. However, to our knowledge, there are no studies that document immunostimulant properties of PAA in Leishmania infection. The main aim of this study was to investigate the interaction of Leishmania parasites with PAA: the possible effects on the infectivity of Leishmania promastigotes; and, induction of nitric oxide (NO) production in macrophages in vitro. The cytotoxicity of PAA on both macrophages and Leishmania infantum promastigotes were determined by MTT assay. NO production in the macrophage culture supernatant was measured by the Griess method. A significant, dose-dependent and time-dependent decrease in the infection index was observed after PAA exposure. The value of this decrease was found to be between 93% and 100% for all concentration and time points. PAA (molecular weight (MW) 30, 100 kDa at 1mg/1h)-exposed parasites stimulate NO production significantly at 48 h post-infection (PI), when compared to the control. This study demonstrates that Leishmania parasites lost their virulence upon interaction with PAA, and this interaction induced NO production in infected macrophages. These results may have important implications in the development of anti-leishmanial vaccines and amelioration of immune response.

Microcapillary culture method: a novel tool for in vitro expansion of stem cells from scarce sources.

BACKGROUND AND AIMS: Although increasing numbers of studies report the derivation of stem cells from a variety of different tissues, derivation efficiencies greatly vary among different studies even for the same tissue source. Hence, a consistent and efficient isolation protocol has not yet been established to date. Several factors have so far been documented that influence and limit mesenchymal stem cell (MSC) isolation and cultivation, including the age and gender of the tissue donor, origin of the tissue, amount of sampled tissue material and cell culture characteristics including the choice of basal media, serum, gas composition, etc. The aim of the study was to investigate the microcapillary culture method (MCM) to establish an efficient and consistent isolation as well as cultivation protocol by comparing the results with other classic culture systems (flasks, center wells). METHODS: MSCs isolated from adipose tissue of different donors were observed comparatively under different culture systems (flasks, center wells, microcapillary tubes) and their proliferation and differentiation were investigated. Flow cytometry was used for immunophenotypic characterization of derived cells and histochemical staining (Oil Red O and Alizarin Red S) was applied for determining their differentiation capacity. RESULTS: It has been shown for the first time that AD-MSCs can consistently and efficiently be derived from a scarce amount of adipose tissue by MCM. CONCLUSIONS: Further and similar studies should be performed to determine whether this methodology can also be applicable for other MSC sources.