Critical Variables in MTT Cytotoxicity Assessment of Liposomal Drug Delivery Systems

Tacio Hayasaki – student, FarmaTec - Laboratory for RD&I in Pharmaceutical Nanotechnology and Drug Delivery Systems, Samambaia Technology Park, Goiania - GO - Brazil; Lucas Sousa – student, FarmaTec - Laboratory for RD&I in Pharmaceutical Nanotechnology and Drug Delivery Systems, Samambaia Technology Park, Goiania - GO - Brazil; Lucas Santos – student, FarmaTec - Laboratory for RD&I in Pharmaceutical Nanotechnology and Drug Delivery Systems, Samambaia Technology Park, Goiania - GO - Brazil; Sebastião Mendanha – Professor, FarmaTec - Laboratory for RD&I in Pharmaceutical Nanotechnology and Drug Delivery Systems, Samambaia Technology Park, Goiania - GO - Brazil; Eliana Lima – Professor, FarmaTec - Laboratory for RD&I in Pharmaceutical Nanotechnology and Drug Delivery Systems, Samambaia Technology Park, Goiania - GO - Brazil

Introduction: Liposomes have profoundly impacted drug-delivery science by enabling encapsulation and controlled distribution of lipophilic and hydrophilic drugs, leading to enhanced pharmacokinetics and improved safety profiles. In drug development, in vitro cytotoxicity assays are essential for dose selection and toxicity screening. The MTT assay is a gold-standard method for free drugs, supported by standardized protocols. However, its application to liposomal systems requires careful assay design and validation. Using miltefosine-loaded liposomes, we evaluated how formulation characteristics influence MTT-based cytotoxicity and identified key parameters for accurate in vitro assessment.