The project aims to develop a novel method to fabricate homogenous and non-aggregated nano-sized hydroxyapatite (HA) particles capable of delivering therapeutic nucleic acids into the cell nucleus. We will:
• fabricate HA nanoparticles using vortex mixture method and characterise their size distribution, and physicochemical and morphological properties;
• load plasmid DNA (pDNA) onto HA nanoparticles and determine the effects of particle characteristics on HA/DNA complexing efficiency, DNA stability, and the in vitro DNA release profile from the particles;
• determine the cytotoxicity and transfection efficiency of HA/DNA complexes;
• functionalise the surface of HA nanoparticles and assess their effects on DNA complexation and transfection efficiency
Gene therapy is a medical technology that has widely recognised potential for treating, preventing, and retarding a wide range of inherited and acquired human diseases. Using this technique, a therapeutic nucleic acid is transfected into the cell nucleus to modulate the synthesis of specific proteins or enzymes associated with certain diseases, including cancers, infection, and genetic disorders. The principle challenge of gene therapy is in the effective and safe delivery of the therapeutic nucleic acids into human cells.
Viruses are efficient delivery vectors for transfection due to their natural ability to deliver genetic material into the nucleus of a cell. However, the clinical application of these viral vectors has encountered serious problems. A synthetic non-viral vector in the form of an organic or inorganic nanoparticle could offer an attractive alternative. Unfortunately, cytotoxicity and low in vivo transfection efficiency presently precludes the use of non-viral vectors in gene therapy.
Studies have demonstrated high transfection efficiency of nano-sized calcium phosphate (CaP) vectors compared to other non-viral gene carriers. CaP in the form of hydroxyapatite and tricalcium phosphate have a proven track record of biocompatibility as bone graft biomaterials. The calcium ion of the CaP vector protects the therapeutic genes against both extra- and intra-cellular degradation and facilitates nucleic acid uptake into the nucleus of the targeted cells. The key challenge in using CaP vectors is the synthesis of non-aggregated and homogenous particles, within the nanometer size range (50 to 200 nm), under conditions that allow the complexation of therapeutic genes with the CaP vector.
The application of CaP nanoparticles in bone tissue engineering and remineralization of dental caries will be also investigated in this project. The project will collaborate with Biomaterials unit, Faculty of Dentistry, University of Sydney
The opportunity ID for this research opportunity is 1721