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(Cluj-Napoca) The effect of metallic nanoparticles functionalized with natural extracts on oral carcinogenesis. In vitro and in vivo study
Romania (FASMR) - University of Medicine and Pharmacy "Iuliu Hatieganu", Cluj-Napoca
Department of Physiology, University of Medicine and Pharmacy “Iuliu Hatieganu”, Cluj-Napoca, Romania
Professor Doctor Simona Clichici
Professor Doctor Simona Clichici, Professor Doctor Filip Gabriela
Type of Research Project
- Basic science
What is the background of the project?
In recent years, the interest in herbal medicinal or fruits products, especially in the field of chemoprevention, has been raised enormously. Due to their poor solubility or a short half-life the incorporation in nanostructures can offer excellent platforms for delivering compounds to the desired site and targeted therapy. While scientists have been able to fine-tune and engineer the properties of nanoparticles by changing their size, shape, surface chemistry and even physical state, such a variety of possibilities means that dictating precisely how the particles behave at that small scale also becomes extremely difficult. This is of particular concern as we rely on the potential use of nanoparticles within the human body. Metallic nanoparticles are good carriers of large and small molecules, making them ideal for transporting drugs to human cells.Several studies demonstrated that silver nanoparticles (AgNPs) have remarkable free radical scavenging and anti-inflammatory properties and also antitumor effect. However, the interactions between AgNPs, natural compounds and tissues are complex, therefore careful evaluation of real efficacy and safety is a prerequisite for the development of a specific chemopreventive product.
What is the aim of the project?
The aim is to evaluate the applicability of metallic nanoparticles in chemoprevention by using in vitro and in vivo studies.
What techniques and methods are used?
This in vitro study includes the testing of various concentrations of silver nanoparticles and natural products respectively on normal cells (fibroblasts) and on dysplastic cells from oral mucosa. First of all, the effect will be quantified by evaluation of viability, oxidative stress markers (malondialdehyde, glutathione reduced/oxidized ratio, carbonyls protein and antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase). Oxidative stress is defined as the imbalance in the redox characteristics of some cellular environment which can be the result of either biochemical processes leading to the production of reactive species, exposure to damaging agents. The direct quantification of reactive oxygen and nitrogen species (ROS/RNS) will be assessed with mass spectrometry techniques, targeting the cell cultures both collected from serum and directly from tumours. Spectrofluorimetry methods will be used in order to analyse the inflamed tissue. Inflammation markers such as interleukin -6 and tumor necrosis factor alpha will be evaluated through the enzyme-linked immunosorbent assay (ELISA).This is a commonly used analytical biochemistry assay that uses a solid-phase enzyme immunoassay (EIA) to detect the presence of a ligand (commonly a protein) in a liquid sample using antibodies directed against the protein to be measured. The method of Real-time quantitative Polymerase Chain Reaction will be used to identify the apoptotic genes p53, Bcl-2, bax. The technique allows the sensitive, specific and reproducible quantitation of nucleic acids that encode the genes . For in vivo study, the oral carcinogenesis induced by chemical carcinogen topical application (4-nitroquinoline 1-oxide) will be used. The effects will be evaluated by oxidative stress markers and cytokine secretion and also by quantification of apoptosis.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The students will learn how to manipulate the laboratory animals such as laboratory rats, to anesthetize them, to take samples such as mucosa or biopsies from tumors. They will also learn how to prepare de nanoparticles. The selection of appropriate method for the preparation of nanoparticles depends on the physicochemical character of the polymer and the drug to be loaded. The emulsification-solvent evaporation technique involves two steps. The first step requires emulsification of the polymer solution into an aqueous phase. During the second step polymer solvent is evaporated, inducing polymer precipitation as nanospheres. The nanoparticles are collected by ultracentrifugation and washed with distilled water to remove stabilizer residue or any free drug and lyophilized for storage. The created nanoparticles will be afterwards used as treatment for the tumors, both in vitro and in vivo. The students will be under the supervision of the tutor.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Theoretical teaching will be provided by the tutor and supervisor. The students will receive some lectures for a better understanding of the project.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No specific skills required
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system
- Xi-Feng Zhang; Zhi-Guo Liu; Wei Shen and Sangiliyandi Gurunathan. SILVER NANOPARTICLES: SYNTHESIS; CHARACTERIZATION; PROPERTIES; APPLICATIONS AND THERAPEUTIC APPROACHES. International Journal of Molecular Sciences 2016 Sep 13; 17(9). pii: E1534. doi: 10.3390/ijms17091534
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