Medical imaging represents a cornerstone in the early detection, diagnosis, and monitoring of cancer progression, playing a crucial role in guiding therapeutic decisions. Despite continuous advancements, conventional imaging techniques still face significant challenges related to sensitivity and specificity, which can lead to diagnostic inaccuracies. One of the most promising strategies to overcome these limitations involves the use of nanotechnology-based imaging agents, which exploit the Enhanced Permeation and Retention (EPR) effect to achieve selective tumour targeting and improved imaging contrast. By leveraging nanoscale properties, these agents enhance the accumulation of contrast materials within tumour tissues, thereby increasing the resolution and accuracy of imaging modalities. This study presents the development and characterization of an innovative nanosystem specifically designed for positron emission tomography (PET) imaging. The system is based on a self-assembling amphiphilic dendrimer that has been functionalized with Gallium-68, a widely used positron-emitting radionuclide known for its excellent imaging capabilities. This dendrimer spontaneously forms highly uniform nanomicelles in aqueous environments, enabling efficient tumour targeting. The ability of these nanomicelles to accumulate preferentially in malignant tissues significantly enhances the sensitivity and specificity of PET imaging, addressing key limitations of conventional imaging agents. A major focus of this research is the detailed investigation of the behaviour of this nanosystem in the bloodstream, with particular attention to its interactions with human serum albumin (HSA), the most abundant plasma protein. Upon entering the circulatory system, nanocarriers often undergo a process known as protein corona (PC) formation, where plasma proteins adsorb onto their surface. This phenomenon can profoundly influence the physicochemical properties, stability, biodistribution, and cellular uptake of nanocarriers, ultimately affecting their in vivo performance. Understanding these interactions is therefore crucial for optimizing the efficacy and safety of nanomedicine-based imaging systems. This work integrates key principles from Biomedical Engineering, Bioengineering, and Bionanotechnology to enhance the development of advanced imaging tools. The application of Biomaterials plays a crucial role in designing the functionalized dendrimer, ensuring optimal biocompatibility and performance in biological environments. Furthermore, Bioprocessing methodologies were employed to synthesize and characterize the nanosystem, ensuring reproducibility and efficiency in its fabrication. To elucidate the interaction dynamics between the dendrimeric nanocarrier and HSA, a comprehensive suite of spectroscopic and calorimetric techniques was employed. Circular Dichroism (CD) spectroscopy was used to assess potential conformational changes in HSA upon binding, while UV-Vis and Fluorescence Spectroscopy provided insights into the binding affinity and quenching mechanisms. Furthermore, Isothermal Titration Calorimetry (ITC) was utilized to determine the thermodynamic parameters governing the interaction, allowing for a deeper understanding of the binding forces involved in protein corona formation. The experimental findings reveal a strong interaction between the functionalized dendrimer and HSA, leading to the formation of a stable protein corona. This interaction is characterized by specific thermodynamic signatures that suggest a combination of hydrophobic and electrostatic forces playing a key role in the binding process. The formation of this stable PC has important implications for the pharmacokinetics and biodistribution of the nanosystem, influencing its circulation time and potential targeting efficiency. These insights are essential for the rational design and further optimization of nanocarriers intended for biomedical imaging applications. In conclusion, the results of this study contribute valuable knowledge to the field of nanotechnology-enhanced medical imaging, providing a deeper understanding of the interplay between nanocarriers and biological environments. The findings underscore the potential of amphiphilic dendrimer-based nanosystems as next-generation PET imaging agents, offering improved diagnostic accuracy and tumour specificity. Future research will focus on in vivo validation and further refinement of these nanocarriers to enhance their clinical applicability in precision cancer diagnostics. The integration of Molecular, Cellular, and Tissue Engineering principles will be crucial for optimizing the interactions between nanocarriers and biological systems. Moreover, Drug Delivery strategies will be explored to extend the application of this nanosystem beyond imaging, potentially enabling theranostic applications. By advancing the development of highly specific and sensitive imaging tools, this work represents a step forward in the integration of nanotechnology with molecular imaging for improved patient outcomes
