13^th International Conference and Exhibition on Nanomedicine and Pharmaceutical Nanotechnology July 24-25, 2017 Rome, Italy

Javier Batista Perez has his expertise in biophysical characterization of protein-protein interaction. His extensive experience in building customized surface chemistry for surface plasmon resonance imaging (SPRi) screening allowed him to design novel SPRi methods to study antibody-antigen interactions in complex samples such as cell lysate. Currently, he is acquiring experience in high-throughput biological screening and DNA sequencing. He is working on a project that intends to develop a more reliable diagnostic method for gout as well as other crystal arthropathies

Crystal arthropathies, as Gout, are often challenging to diagnose and are frequently mismanaged, in part due to the lack of sensitive and reliable diagnostics. Optical microscopy of monosodium urate monohydrate crystals (MSU) is the gold-standard diagnostic approach, but is cumbersome and often not performed, and has poor sensitivity and high inter-operator variability. Consequently, there is a pressing need to develop new diagnostic strategies for Gout, leading to improved patient outcomes and enabling new research tools that will aid our understanding of the complex pathogenesis of this condition. On this research peptides that bind to MSU were identified by medium of a combination of phage display screening, next generation sequencing and peptide chemistry. Furthermore, a novel method to detect the presence of MSU, based on its interaction with gold nanoparticles coated with the identified peptides, was developed. We consider that the method developed here can be used as new diagnostic method for Gout which will help to a more precise diagnosis of this condition.

Silvia Mellace is a PhD student at the University of Calabria. Her PhD is in Translational Medicine, with the main research interests in design and characterization of novel micro/nano drug delivery systems for the improvement of therapeutics delivery. In particular, her studies are focused on formulations for topical application. Currently, she is doing an internship at the School of Pharmacy, University College of Cork, investigating in vitro and in silico methods for the effective prediction of drug dermatopharmacokinetics using dissolvable microneedles arrays.

In the last decade, a number of different drug delivery systems have been formulated to enhance the transport of therapeutics across the skin barrier. In this study, we combined dissolvable microneedles (DMN) and liposomes as simple and minimally invasive strategy to deliver free or nanoencapsulated drug into or across the skin. Physicochemical properties of liposomes, in particular particle surface charge and size, have a significant effect on their permeation and subsequent distribution through microchannels created in the skin by DMN. Hence, the main focus of this work was to assess the effects that microneedle fabrication process may have on liposome characteristics and stability. Ketoprofen-loaded liposomes were formulated using thin film hydration, extrusion and lyophilisation methods. Micromolding technique was used to fabricate DMN containing free ketoprofen and ketoprofen-loaded liposomes. The vesicles surface charge and size were evaluated before and after DMN fabrication and used as a measure of the liposomes ability to maintain their structure during stress conditions of DMN fabrication process. The results revealed that ketoprofen-loaded liposomes can be successfully incorporated within DMN and used for microneedle-mediated intradermal and transdermal drug delivery. The rigorous processing conditions during DMN fabrication didn’t affect liposomes characteristics. However, the amount of drug that could be delivered using DMN when ketoprofen is encapsulated in liposomes is probably not sufficient to achieve a therapeutic effect in comparison to free drug, despite a high ketoprofen encapsulation efficiency of 83.9%. Further work is required to improve the loading efficiency of microneedles containing liposomal nanoencapsulated drug and to understand the behaviour of these formulations within the biological environment. Our findings contribute to the further development of effective, painless and minimally invasive technologies for intradermal and transdermal drug delivery.

Neeranut Kuanchertchoo has her expertise in waste utilizations and applications in biomedical materials as well as water purification. Her research emphasizes on development of nanofiber membranes using electrospinning technique. Her laboratory skills are synthesis, characterization and application in biomaterials, separations and purification. She earns many years experiences in both teaching and performing researches.

Fruit peels contain many useful substances which can be used as antioxidant, antimicrobials, and antivirus, etc. In this work, Punica granatum peels were cleaned, extracted with water and freeze dried into powder. The crude extracts were characterized using FTIR. The spectra represented the peaks at 3,271 cm-1 (OH stretching), 1,610 cm-1 (N-C=O stretching of amine groups) and 1,375 cm-1 (C=O stretching of ketone or aldehyde) of polyphenol. Antibacterial activities of water crude extracts were also evaluated by agar diffusion method. It was found that Staphylococcus aureus ATCC 25923, Staphylococcus aureus (MRSA) DMST 20654, Staphylococcus epidermidis ATCC 12228 and Acinetobacter lwoffii ATCC 15309 were inhibited. Crude extracts from Punica granatum peels showed high efficiency to be used as natural antibacterial agent with low cost. For application, polyvinyl alcohol and chitosan solution (20:1 in volume) were mixed with crude extracts from Punica granatum peels and formed into nanofiber using electrospinning technique. SEM micrograph showed structure of nanofiber with some beads. Polyvinyl alcohol and chitosan naofiber trends to be a suitable matrix for releasing of Punica granatum peel extracts and applied as wound dressing.

Young Bong Kim received his Doctorate from Sogang University in Korea and trained at the NIAID/NIH in the United States. Since his appointment as a Professor at Konkuk University in 2003, he has been working on several vaccines against pathogenic viruses such as HIV, MERS-CoV and ZIKA virus.

Middle East respiratory syndrome Coronavirus (MERS-CoV) has emerged as a new pathogen that can transmit between humans as well as animals and humans, causing severe complications and high mortality rates. MERS-CoV continues to spread throughout the pandemic spread around the globe. MERS was first discovered at the end of 2012 and has caused more than 1,800 infections and 650 deaths. Increased MERS cases and no licensed MERS vaccines highlight the need for safe and effective vaccine development for MERS. The MERS-CoV spike (S) protein is responsible for receptor binding and virion entry into the cell and is highly immunogenic and induces neutralizing antibodies. In this study, we have expressed the eS1-770 MERS-CoV Spike protein fused with human Fc4 (eS1-770-Fc4) using baculovirus system and purified. Different doses of the eS1-770-Fc4 vaccine candidate were intramuscular injected into mice, and blood samples were collected every 10 days after immunization. The eS1-770-Fc immunized sera showed high titers of MERS-Cov antibodies and neutralizing activity against MERS-CoV. Only 1 ug of eS1-770-Fc4 elicited enough immunogenicity without adjuvant. The eS1-770-Fc could be a potential vaccine candidate.

Badriah Hifni received her Master’s degree at King Abdulaziz University of Jeddah (Saudi Arabia) for her work on the genotoxicity and cytotoxicity of carboplatin in somatic and germ cells of adult male swiss albino mice under the supervision of Dr. Salwa Mohammad. Currently, she is in her third year of Doctor of Philosophy (PhD) program at University College Dublin. The focus of her PhD is the investigation of the cellular uptake of functionalized carbon nanoparticles under the supervision of Prof. Jeremy Simpson and Dr. Susan Quinn.

Nanomaterials possess excellent structural, electronic and optical properties and are seen to offer huge potential in nanomedicine applications. Due to their biocompatible chemical composition, carbon nanoparticles have received significant attention in recent years, yet many fundamental questions still remain about their mechanism of internalisation by cells. The aim of this project is to address this lack of knowledge for two forms of carbon particles, carbon nanohorns (CNHs) with high surface area and chemical inertness, and carbon nanodots (C-dots) that display good water dispersibility, chemical inertness and high biocompatibility. A range of chemical synthesis methods were used to generate C-dots. These were characterised using a range of techniques including AFM, DLS and FTIR and their cellular uptake was assessed by utilising in vitro HeLa cell-based assays to determine how internalisation occurs and the pathways they follow in cells. Three families of C-dots have been successfully synthesised by three different approaches, namely microwave, electrochemical and solvothermal synthesis. The resulting C-dots showed emission profiles in the blue, green and red parts of the spectrum under different excitation conditions. A maximum emission peak at 500 nm is observed when the green C-dots are excited at 440 nm, whereas the CNHs emit at 568 nm when excited at 551 nm. Upon characterisation, AFM and DLS methods determined the average diameter of the C-dots and CNHs to be ~ 4 nm and 198 nm respectively. MTT cytotoxicity assays suggest that the blue and red C-dots do not induce any significant toxicity to cells, while the green C-dots at 25 µg/mL cause an approximately 60% reduction in cell viability. Overall, different classes of carbon nanoparticles with low cytotoxicity can be readily synthesised and subsequently utilised to study their trafficking in mammanlian cells. Future work will focus on unravelling the specific pathways involved in this trafficking process with a view to better designing carbon nanomaterials to act as vehicles for drug delivery.

Mona Kabiri is a PhD Candidate of Pharmaceutical Nanotechnogy at School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. She is a Guest Researcher at Institute of Experimental Hematology and Transfusion Medicine, University of Bonn, Bonn, Germany. She finished her PhD thesis about protein vaccine delivery system based on PLGA polymeric nanoparticles and nano-adjuvants. She has published three articles in ISI journals and has three national and US-patents.

Among the novel delivery systems, polymeric nanoparticles have been considered as promising carriers for therapeutic applications. The use of biocompatible and biodegradable polymers such as polylactic-co-glycolic acid (PLGA) as protein and drug carriers has long been of interest in controlled-release technology. PLGA nanoparticles act as potential carriers for several classes of lipophilic and hydrophilic drugs such as anticancer agents, immunomodulators and hormones, as well as macromolecules including nucleic acids, proteins, peptides, and antibodies. The PLGA nanoparticles were prepared by water-in-oil-in-water (w/o/w) (double emulsion) method. In this method, the aqueous protein solution (BSA) was added to the PLGA organic solution. The mentioned method is best suited to encapsulate water-soluble drugs like peptides, proteins and vaccines. The nanoparticles were characterized in terms of particle size, poly dispersity index (PDI) and zeta potential, as well as entrapment efficiency of I-125 labeled BSA. Furthermore, particle morphology and size are determined with scanning electron microscopy (SEM). As estimated by dynamic light scattering (DLS), the mean size of the PLGA nanoparticles were about 150 nm and 185 nm (n=5) for blank PLGA and PLGA-BSA after lyophilization, respectively. Also, The DLS analysis indicated a unimodal particle size distribution, with PDI of 0.2 and zeta potential of -38 mV. PLGA polymers were non porous, smooth surfaced and spherical in structure under SEM with a mean particle size ranging from 70 to 100 nm. The loading efficiency of the radiolabeled protein in PLGA nanoparticles was 91.2%. Radiolabeled protein encapsulated in PLGA nanoparticles can be useful for therapeutic projects.

Yuan-Chia Chang is a senior researcher of Biomedical Technology and Device Research Laboratories (BDL), Industrial Technology Research Institute (ITRI), Tawan, R.O.C. ITRI is a nonprofit R&D organization engaging in applied research and technical services. The vision of BDL is to become a premier global hub for the research and development of innovation-based biomedical technology and medical devices. Dr. Chang graduated from Chang Gung University in Medical Technology, obtained his MS degree in Biomedical Engineering department from National Yang-Ming University, and obtained his PhD degree in Chemical Engineering from National Tsing-Hua University in Taiwan, R.O.C. He majors and specialties are focused on bio-polymer application and drug delivery system.

Head and neck carcinoma is the sixth most common cancer worldwide. 50-60% of patients with stage III or IV disease present locoregional recurrence within 2 years. Given the limitations of surgical resection and the low effectiveness of radiotherapy and chemotherapy, the treatment of lymphatic metastatic tumors remains a great challenge. Nanoparticles are potential tools for the lymphatic drug delivery but the major hurdle of existing nanoparticles is poorly retained in the draining lymph nodes following injection (less than 2% injected dose). Hyaluronic acid (HA) is seen as an ideal carrier for the treatment of lymphatic tumors due to it is the ligand for CD44 receptor and cleared primarily by the lymphatic system. In this work, we performed an improved Cisplatin-incorporated HA nanocomplexes (CPHC008) and evaluated the potential use as carriers for lymphatic drug delivery and tumor lymphatic metastasis inhibition. HA/Boc-His/PEG graft copolymers were mixed with Cisplatin (CDDP) followed by stirring. The mixture was sonicated and purified by ultrafiltration and 0.22 μm filtration to form the CDDP incorporated HA nanocomplexes (CPHC008). The size and zeta potential were 200 nm and -30 mV respectively. Transmission electron microscopy showed the formation of spherical nanoparticles. The drug release behavior from CPHC008 in PBS showed more stable than native HANP. In vitro cellular binding/uptake confirmed the CPHC008 had specificity target the CD44 positive cells. As a result of active targeting properties, CPHC008 could increase platinum levels in the cervical lymph nodes and against lymph node metastases was significant. Taken together, we have designed a HA-based nanocomplexes to increase platinum levels in the lymphatics, where early metastasis is most likely to occur. This lymphatic delivery platform may offer significant advantages for the use of platinum medicines in the management of locally carcinoma, treating microscopic lymph node disease with better efficacy than current high-dose systemic chemotherapy.

Safoura Karami is Master of Medical Toxicology candidate in Pharmaceutical Sciences Branch, Islamic Azad University, Faculty of Pharmacy. Her thesis is effect of N-acetylcystein loaded niosomes on indicators of liver toxicity in male Wistar rats were poisoned with acetaminophen, under supervision of Prof. Amir Nili-Ahmadabadi.

Aim: Acetaminophen (APAP)-induced liver damage is an important health problem and represents the most frequent cause of drug-induced liver failure in the world. N-acetylcysteine (NAC), a sulfhydryl donor is considered the antidote of choice for APAP overdose. The aim of this study is to improve the therapeutic efficacy of NAC in APAP-induced liver injury using niosomes, as a nanocarrier. Methods: Nanoparticles were synthesized using reverse phase evaporation technique and characterized for shape morphology, zeta potential, particle size, and drug-release properties. In the part of in vivo study, male wistar rats were divided into seven groups. The four groups were treated with saline, empty niosomes, NAC and niosomes-NAC, respectively, as control groups. The other three groups were challenged with an oral dose of APAP (2 g/kg); then normal saline, NAC and niosome-NAC were administered to the animal 4 hours later. These animals were sacrificed 48 h post-APAP treatment. Finally, liver function and oxidative stress biomarkers were assayed in tissue and serum samples. Results: APAP administration resulted in hepatic damage as observed by increases in serum aminotransferase enzymes and hepatic tissue levels of NO, LPO as well as decreases in TAC and TTM. Treatment of animals with niosomes-NAC was remarkable more effective than free NAC in reducing serum aminotransferase enzymes; ALT (P<0.05) and AST (P<0.01). In addition, these changes were well related with the decreasing of oxidative damages in liver tissue. Conclusion: The efficiency of Niosomal NAC was more than free NAC in decreasing of APAP-related negative changes; therefore niosome-NAC may have further impact in the treatment of hepatotoxicity induced by APAP.

Alia Moosavian is an Assistant Professor of Pharmaceutical Nanotechnology at Mashhhad University of Medical Sciences. Her work focuses specifically on the drug delivery for cancer treatment. Her main areas of research interest are liposomal drug delivery systems and aptamers as new targeting agents for drug delivery.

Employing targeting ligands with high affinity to tumor receptors is an important strategy to increase treatment efficacy. The use of aptamers as targeting agent is increasingly prevalent in drug delivery systems. Mucin1 (MUC1) is a glycoprotein that is over-expressed on the surface several cancer cells and plays important role in metastasis and invasion. 5TR1-aptamer is a DNA aptamer, which targets MUC1 receptors. We investigated the anti-tumor activity and therapeutic effectiveness of 5TR1-aptamer-PEGylated liposomal Doxorubicin (PLD) delivery system in C26 tumor-bearing mice. The in vitro experiments demonstrated enhanced cytotoxicity and cellular uptake of PLD at the presence of 5TR1 aptamer into MUC1+C26 cell line. Biodistribution study indicated that aptamer conjugation increased tumor accumulation of PLDs. Pharmacokinetic analysis showed despite higher clearance rate, selective delivery of doxorubicin to tumor tissue was increased in 5TR1- Doxil group. In C26 bearing tumor mice, treatment with 5TR1-Doxil exhibited significant deceleration in tumor growth and enhanced survival. The results suggested that 5TR1 aptamer is promising ligand for active targeting which improves therapeutic efficiency of PLD in cancer therapy.

Jocelyn L Miranda is a Professor in the field of Pharmaceutical Practice. She plans to further continue her studies by taking up PharmD in 2018. Her expertise has rooted from years of construing journal and performing laboratory experiments related to microbiological and pharmaceutical research. Her drive to suppress the spread of mosquito-borne viral diseases by eliminating the vectors emanates from a previous dengue infection that almost took her life. As a Licensed Pharmacist, she saw the need to come up with a new larvicidal agent that is easy to prepare, effective and environmentally safe. Lately, nano formulated products has caught her attention and eventually urged her to develop this study. She has worked with her mentor Maria Cleofe Badang, a Chemist, to have deeper understanding of nanoparticles’ physical and chemical characteristics. Results obtained from this study present an environment friendly insecticide that can serve as alternative for traditional mosquito larvae control worldwide.

Mosquito-borne diseases have been a perennial problem in tropical countries. The outbreak of malaria in the 1950’s resulted to the massive use of DDT insecticide, which was eventually banned due to its adverse effects on humans and the environment. In recent years, the emergence of mosquito-borne diseases such as dengue fever, chikungunya and zika viral infections caught worldwide concern and prompted the WHO to address this problem from its root cause – the eradication of the vector. These diseases were vectored by the same type of mosquito, the Aedes aegypti. The aim of this study was to investigate the larvicidal potential of green-synthesized silver nanoparticles (AgNPs) using leaf extract of Cracker Plant (Ruellia tuberosa) against III and IV instar larvae of A. aegypti. Synthesis of the AgNPs were initially detected within 5 minutes after mixing of the extract and 1 mM AgNO3 solution, with optimum reaction time of 12 h as determined by UV-Visible Spectrophotometry. The spectral wavelength shift from 320 nm to 420 nm due to Surface Plasmon Resonance (SPR) proved the formation of AgNPs. Phytochemicals in Cracker Plant served as capping and stabilizing agents in the AgNPs formation. Scanning Electron Microscopy (SEM) revealed synthesized AgNPs to be spherical and oval in shape with sizes ranging from 28-50 nanometers. Fourier Transform Infrared Spectroscopy (FTIR) showed functional groups associated with flavonoids such as luteolin and triterpenoids such as lupeol and betulin. Larvicidal activity was assessed for 24h with varying concentrations of AgNPs. The recorded 90% lethal concentration (LC90) was found to be 105.42 ppm. Bioassay was done using Ovicidal/Larvicidal (OL) pellets and de-ionized water as positive and negative control, respectively. Results showed Cracker Plant AgNPs as larvicidal agent and can be used for mosquito larvae control.

Arianna Mancini has her expertise in drug delivery and nanomedicine in oncology. The principal field of her research is about anticancer compounds, especially pyrazolo[3,4-d]pyrimidines compounds, that represent a valid approach against several cancer types (i.e. glioblastoma, neuroblastoma). Her work covers the preparation and the characterization of pharmaceutical nanotechnologies (i.e. liposomes, albumin nanoparticles, carbon nanodots) and their in vivo administration and pharmacokinetic parameters evaluation. Her knowledge includes elements of biophysics and molecular imaging (magnetic resonance), as instruments for real time monitoring of gadolinium-loaded liposomal nanotechnology.

Pyrazolo [3, 4-d] pyrimidines are a class of compounds with a good antitumor activity against several cancer cell lines. The activity of pyrazolo[3,4-d]pyrimidines has been related to the inhibition of some TKs families, such as Abl and Src. Recently, remarkable results have been obtained in several xenograft mouse models.[2] Despite the promising anticancer activity, these molecules showed a poor aqueous solubility. This issue could limit the future development of pyrazolo[3,4-d]pyrimidines as clinical drug candidates. With the aim of improving solubility profile and consequently the pharmacokinetic properties of our compounds, we have explored albumin nanoparticles and stealth liposomes as possible nanocarrier systems.[1] For this study, we have chosen four compounds (1–4), previously characterized for their activity against neuroblastoma by enzymatic, cytotoxic and in vivo assays.[2-4] For each selected compound (1-4), albumin nanoparticles (AL 1-4) and stealth liposomes (LP 1-4) were prepared and characterized regarding size and ζ-potential distribution, polidispersity index, entrapment efficiency and activity against SH-SY5Y human neuroblastoma cell line. The most promising nanosystem, namely LP-2, was chosen to perform further studies: confocal microscopy, in vitro stability and drug release in physiological conditions, and 24 hours biodistribution test in male Sprague-Dawley rats. Herein we demonstrated that the cytotoxic activity of 1-4 compounds against SH-SY5Y cell line is likely to be exerted by their encapsulation in liposomes, thanks to the release of the active compound in the cellular cytoplasm after the nanoparticles uptake. Importantly, the liposomal system resulted stable in physiological conditions. Finally, the 24 hours biodistribution assay proved the beneficial properties of LP-2 (longer circulation residential time). Further preclinical in vivo studies will allow the determination of the full pharmacokinetic profile and the therapeutic efficacy of this novel formulation.

Alexandra Scheer completed her Master's degree in Medical Biology in 2015. Since October 2015, she has been working on her doctoral thesis at the Institute of Physiological Chemistry at the University Hospital Essen in the working group of Dr. Katja B. Ferenz. Within the scope of this work she is involved in the development and characterization of artificial oxygen carriers.

Statement of the Problem: Although „Patient Blood Management“-programs are implemented in many hospitals1, availability of approved artificial oxygen carriers are missing for clinical use both in Europe and USA2. Therefore albumin-derived perfluorocarbon-based nanocapsules (nanocapsules) were explored as a novel artificial oxygen carrier3,4. Methodology & Theoretical Orientation: To investigate an essential supply of oxygen, we studied our nanocapsules in a model of massive normovolemic hemodilution (NH) followed by an in vivo fluorescence microscopy analysis (IVM) to assess the microcirculation5. NH of 16 healthy wistar rats was performed stepwise below the critical hematocrit of a rat (10%) with nanocapsules and without ones (control), respectively. During NH systemic parameters (e.g. heart rate, mean arterial pressure (MAP), respiration, pH, pO2, lactate) were continuously monitored. At the end of NH rats were further observed up to 100 min and an IVM of the right leteral liver lobe was performed. Vessel diameters of sinusoids, post sinusoidal venules and the number of perfused sinusoidal vessels (%) were determined5. Findings: As expected, nanocapsule-diluted animals showed a higher pO2 and sO2 during the observation period while pCO2 was somewhat lower. At the beginning of the dilution period the nanocapsule-diluted animals showed a higher MAP. The pH and base excess were not affected by hemodilution. Interestingly, the microcirculation was also not affected significantly in the nanocapsules diluted animals. Conclusion & Significance: The results illustrate that the microcirculation was not affected in both groups. Likewise, the parameters essential for survival (e.g. pO2, pCO2) were improved in the nanocapsule-diluted animals. Based on the encouraging results, the nanocapsules will be examined in further clinically relevant settings.