In vitro studies of the fate of advanced polymeric carriers and the formation of a protein corona on their surface

Nanomedicine is a huge area of opportunity for pharmaceutical companies and the biotechnology industry. Modern nanocarriers now make it possible to encapsulate water-insoluble hydrophobic drugs and intractable compounds that need to be protected from blood enzymes such as esterases or nucleases. Polymeric nanocarriers significantly improve the therapeutic index of drugs by increasing their accumulation in specific organs, tissues or cells while reducing their toxicity and side effects. However, a successful transition from laboratory to clinical studies requires a proper evaluation of the interaction of carriers with biomolecules and cells in vivo. It is essential to know the kinetics of drug release and the degradation modes of the nanocarrier both in vitro and in vivo. In this project, we will investigate the biological fate, including in vivo stability, degradation kinetics and biodistribution of materials based on polylactide/poly(lactic-co-glycolic acid) nanoparticles functionalized with polyamines and phosphates. A variety of imaging techniques will be used. The intracellular localisation, nanoparticle aggregation, conjugate stability and drug release mechanism will first be investigated in vitro using advanced techniques such as confocal microscopy, flow cytometry, Raman spectroscopy and fluorescence correlation spectroscopy. The research will be carried out using fluorescence correlation spectroscopy. We also intend to investigate the effect of the generated shells on the polymer on their 'masking' and the formation of a protein corona on them. In order to study the biodistribution of nanoparticles and conjugates with 'payloads', we will further investigate the radio-labelling of the obtained materials, the localisation of which will be determined in vivo using ultrasensitive molecular imaging methods that allow real-time observation of nanoparticles, such as computed tomography (SPECT) and positron emission tomography (PET).The research carried out in this project will contribute to the understanding of the complex interactions of nanoparticles with biological systems. The project will answer fundamental questions about the stability of nanomaterials and their conjugates with drugs in biological systems at the cellular level and under in vivo conditions. The results of the work will be published in peer-reviewed scientific journals and presented at international scientific conferences.The proposed research solutions are unique and provide a novel way to assess the suitability of nanomaterials for medical applications. The methods developed in the project will contribute to the development of tools to support the development of nanomedicine and the rational design and production of nanomaterials.