NanoSystems

In the last years, nanotechnology is emerging as a field of interest for research and industry. Nanotechnologies include all the emerging technologies dealing with the manipulation of matter at the atomic and molecular level, specifically with the design, the characterization and the synthesis of nanomaterials (NM). NMs can be applied in many fields including medicine, agriculture, food safety, cosmetics and industrial applications such as sensors and catalysts. The interest in nanostructured materials comes from the fact that they exhibit properties and functions that differ from bulk-size materials. At the nanoscale the increase of the surface per unit mass results in a higher chemical reactivity, higher resistance to mechanical stress and an alteration of the electrical properties, such as an increase of conductivity.

Magnetic NPs (MNPs), in particular iron oxide nanoparticles (IONPs), are a class of NPs with the potential to revolutionize the biomedical field thanks to their small size, comparable to those of biological molecules, and their physical properties, like superparamagnetism and high coercivity, which allow their manipulation with an external magnetic field. Another important feature of IONPs is the presence of a large surface that can be coated with molecules that stabilize and protect the surface from biodegradation, subsequently allowing the conjugation of different biological molecules such us enzymes, antibiotics or drugs.

 

THERMAL TUNING OF MULTI ENZYMATIC CASCADES (HOTZYMES PROJECT)

Metal-based nanoparticles are used in very diverse fields ranging from environmental remediation to magnetic resonance imaging, targeted therapy and multi-enzyme processes. This last application has been exploited in the European project (FET-OPEN HOTZYMES project; https://www.hotzymes.eu) of which our lab is an active partner. This project involves the remote and tunable activation of enzymatic processes, producing several novel metal-based nanoparticles with a broad spectrum of magnetic characteristics and surface functionalization for the optimization of enzymatic binding.
Our Laboratory is involved in work packages aimed to assess the stability and safety of these newly synthesized nanoparticles. In particular, for what concern the safety we have focus our attention to epigenetic modifications using NIH-3T3 mouse embryo fibroblast cell lines as model. 

METHODS:

  • Cytotoxicity: cell viability of NIH-3T3 cells line, exposed to different nano systems, is assessed by ATP CellTiter Glo assay.
  • Epigenetic modifications are investigated by Chromatin immunoprecipitation (ChIP) assay for the histone modifications anti-H3, anti-H3K27ac, anti-H3K4me, anti-H3K27me3, anti-H3K9me2 and anti-H3K9me3 and then Genome-wide DNA sequencing (ChIP-seq).

 

NANO- ANTIBIOTIC

One of the aims of the research conducted in our Lab is the development of a nano- antibiotic system, using IONPs as a platform for the conjugation of two glycopeptide antibiotics: teicoplanin and vancomycin. In the era of antimicrobial resistance, the use of nanoconjugated antibiotics is regarded as a promising approach for preventing and fighting infections caused by resistant bacteria thanks to the possibility to direct conjugated antibiotics to infection sites, facilitating tissue penetration and disturbing biofilm formation, with a concomitant reduction of side effects and resistance.

METHODS:

  • Synthesis of IONPs by a co-precipitation method.
  • Functionalization with (3-aminopropyl) triethoxysilane (APTES): to allow the presence of functional groups (-OH or -NH2) exposed on their
  • Conjugation of vancomycin and teicoplanin.
  • NPs characterization by TEM analysis and evaluation of dissolution.
  • Evaluation of antimicrobial efficacy through classical microbiological methods (i.e. growth kinetic analysis, minimal bacteriostatic and bactericidal concentrations and agar diffusion assay).
  • Cytotoxicity of nano-antibiotic on different cells line (i.e. NIH-3T3, SKOV-3).

Department of biotechnology and Life Science – DBSV