Multi-step In Vitro Hemocompatibility Testing Protocol Recapitulating the Foreign Body Reaction to Nanocarriers
Résumé
The development of drug nanocarriers based on polymeric, lipidic and ceramic biomaterials has been paving the way to precision medicine where the delivery of poorly soluble active compounds and personalised doses are made possible. However, the nano-size character of these carriers has been demonstrated to have the potential to elicit pathways of the host response different from those of the same biomaterials when engineered as larger size implants. Therefore, a specific regulatory framework needs to be made available that can offer robust scientific insights and provide safety data by reliable tests of these novel nano-devices. In this context, the present work presents a multi-step protocol for the in vitro assessment of the hemocompatibility of nanocarriers of different physicochemical properties. Poly(EthylButyl Cyanoacrylate) nanoparticles and lipid-based (LipImageTM 815) nanoparticles of comparable hydrodynamic diameter were tested through a battery of assays using human peripheral blood samples and recapitulating the main pathways of the host response upon systemic administration; i.e. protein interactions, fibrinogen-platelets binding, cytotoxicity and inflammatory response. The data showed the sensitivity and reproducibility of the methods adopted that were also demonstrated to determine individual variability as well as to discriminate between activation of pathways of inflammation and unintended release of inflammatory signalling caused by loss of cell integrity. Therefore, this multi-step testing is proposed as a reliable protocol for NP development and emerging regulatory frameworks.