Project title: Development of novel Gold NanoRods Polymeric NanoParticles structures for targeted diagnostic and therapeutic applications
Acronym: GNR-PNP
Coordinator: SiTec consulting

Type: Feasibility study
Annuality: Third
TP/LS membership: CMDI/HMNA 1
Subjects involved: Dirivet, University of Turin, Ephoran Multi Imaging

Status: Completed
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Abstract: Gold NanoRods (GNRs) are elongated gold nanoparticles with distinctive physicochemical characteristics directly related to their shape. GNRs can be used as diagnostic agents in biological imaging, with high sensitivity and contrast efficiency, using various technologies such as dark-field microscopy, optical imaging, and photoacoustic imaging. In this project, SiTec Consulting and Partners (Dirivet SaS; University of Turin; Ephoran Srl ) investigated the developability of innovative nanostructures based on Gold Nanorods (GNR). In-vitro , on cell lines, and in-vivo, the potential application of GNR-PNP in some major oncological areas was evaluated, both as a diagnostic contrast agent and as a possible therapeutic agent based on photothermal therapy with laser irradiation.

Name: Hana Danan, Ph.D
Organization: SiTec Consulting
Phone: +39 3493283430


The overall aim of the GNR-PNP project is to define the developability of innovative nanostructures based on Gold Nanorods (GNRs), to study the ‘efficacy of various ligands, and to evaluate their potential for application in some major oncological therapeutic areas, either as a diagnostic contrast agent or as a therapeutic agent based on photothermal therapy with near-infrared (NIR) laser irradiation. Nanostructures based on GNRs, due to the peculiar plasmonic absorption physical properties of these, enable the combination of diagnostic properties with therapeutic use in a single system (“Theranostics”) and represent one of the potentially most viable application areas of biomedical nanotechnology. A further objective of the project is the exploitation of an innovative chemical synthesis protocol, for the preparation of GNRs in an aqueous environment, which allows for the simple and reproducible production of amphiphilic particle nanostructures less than 100 nm in size, containing GNRs derivatized with different ligands, and stable in an aqueous environment at physiological pH (GNR-PNPs), by a dual phase transfer process. Due to these chemical-physical characteristics,
GNR nanostructures obtainable by the dual phase-transfer protocol are expected to be biocompatible, and easily formulated for systemic and/or localized administrations.


To achieve the project goals, the partners completed the following activities:

  • Strategies for the synthesis of GNRs, and of their incorporation into polymeric nanoparticles, were outlined. In addition, targetting agents were selected.
  • Several cell lines, representative of frequently diagnosed neoplasms, were chosen. Specifically selected were MCF-7, cells derived from metastasis of breast adenocarcinoma; CaCo-2: from colon adenocarcinoma; A549: from lung carcinoma; PC-3: from prostate carcinoma; IGROV-1, cells derived from ovarian carcinoma.
  • Some initial batches of “blank” GNR-PNPs, i.e., without conjugated ligand, were made, characterized for particle size, zeta potential, and elemental gold (Au) content.
  • In vitro Optical Imaging and CT/X-Ray Scan tests were performed on the GNR-PNP samples to assess whether or not the gold loading (of GNR) within the colloidal particles should be increased. A response “baseline” was defined, and the gold loading (of GNR) within the colloidal particles was determined. Based on the initial results , Au loading in the nanoparticles was increased, and a marker for fluorescent OI (cyanine) was bound to the surface of the nanoparticles.
  • The most suitable molecules for targetting were selected, and the synthetic activities of derivatized GNR-PNPs with the appropriate functional groups for conjugation were defined.
  • To evaluate possible cytotoxic effects of the conjugated GNR-PNP constructs, cell proliferation tests, using the MTT viability assay, were performed on the various tumor lines. In addition, cellular internalization (uptake) of nanoparticles was measured, and the effect of specific ligands was evaluated.

A preliminary in vivo study (nude mice) was performed to evaluate the biodistribution of GNRPNP-Cyanine after IV administration. This study confirmed the possibility of monitoring GNR-PNP-Cy in vivo by Optical Imaging (OI). In vivo research activity was focused on identifying the diagnostic modality that would best highlight the potential of GNRs-PNPs constructs. They evaluated themselves:

  • X-ray computed tomography , which exploits the x-ray mass absorption characteristics of gold.
  • Optical Imaging (OI), With OI it was possible to perform distribution studies of nanoparticles both
  • in the healthy animal, both in induced tumor models, and evaluate binding on tumor models that overexpress folate receptors, or fatty acid transport systems.
  • Photo-acoustic imaging, which allowed exploiting the ability of GNRs-PNPs to absorb near-infrared laser radiation by combining optical/NIR images with those obtained by ultrasound scanning


  • The possibility of effectively incorporating GNR into polymeric nanoparticles has been demonstrated, and its synthesis has been modified to conjugate molecules with different chemical and biological characteristics to the surface.
  • Moreover, it has been shown in vitro that such modifications significantly affect cellular uptake
    of the constructs, without inducing relevant toxicity.
  • In vivo, it has been shown that the different imaging techniques now state of the ‘art are definitely applicable in the detection of GNR-PNP systems, and that the potential exists for possible application in cancer therapy.
  • In addition, results obtained by photoacoustic imaging confirm intratumoral uptake of circulating nanoparticles, and suggest the need for dose adjustments, and optimization of the physical characteristics of GNR-PNPs (size and surface charge).
  • Lipid ligands with particular affinity toward specific tumor cell lines have been identified

It is expected that the results obtained in this project, appropriately publicized, will foster entrepreneurial interest in this area, and the genesis of one or more preclinical and possibly clinical development projects, involving a network of academic and industrial partners to add to the current established “core of partners.” Such projects will be able to generate an economic return in the short to medium term, and foster the growth of the businesses involved. In addition, the possibility of using GNR-PNPs for selective eradication of cancer cells with localized therapy will increase diagnostic and therapeutic options in oncology, with a possible improvement in patients’ quality of life.


  • Other private partners: DiriVet SaS, Ephoran Multi Imaging Solutions Srl
  • Other public partners: University of Turin, Faculty of Veterinary Medicine, Department of Pharmacology and Toxicology
  • External Collaborators for SiTec Consulting (outside the Region): Dr. Mauro Comes Franchini, University of Bologna , Faculty of Industrial Chemistry
  • Total number of partners: 4
  • Number of employed researchers (fixed-term and permanent and cocopro) involved: 6
  • Total budget: €157. 270,00
  • Funding: €72. 675,00
  • Number of presentations at conferences and seminars: 2
  • Number of patents filed: 1 patentability evaluation
  • Number of permanent or cocopro jobs created: 1
  • Number of jobs retained at the end of the project: 6
  • Number of public researchers involved: 2