By Nanotechnology Characterization Laboratory (NCL)
A thorough understanding of nanomedicine pharmacology and toxicology is essential to identify liabilities and optimize drug formulation. Pharmacology and toxicology properties are frequently not identified until in vivo preclinical studies are performed later in the development process. There are, however, several predictive in vitro assays that can be extremely informative for development of nanomedicines, such as drug release in physiologically relevant matrices, cytotoxicity in cell lines relevant to a specific indication, and assays to evaluate mechanisms of toxicity. In addition to these in vitro studies, the NCL also provides in vivo pharmacokinetic and toxicity studies of nanomaterials. In vivo studies are conducted in collaboration with the Frederick National Laboratory's Laboratory Animal Sciences Program, which includes the Molecular Histopathology Laboratory and Small Animal Imaging Program, among others. All in vivo studies are individually tailored for each tested nanoparticle and are collaboratively agreed upon with the developer before testing begins; in vivo studies are designed to mimic the intended clinical dose (adjusted for species), dosing regimen, and route of administration. In Vitro Cytotoxicity (general): GTA-1 MTT and lactate dehydrogenase release in porcine renal proximal tubule cells; GTA-2 MTT & LDH release in human hepatocarcinoma cells. In Vitro Cytotoxicity (apoptosis): GTA-5 Caspase 3 activation in porcine renal proximal tubule cells; GTA-6 Caspase 3 activation in human hepatocarcinoma cells; GTA-14 Caspase 3/7 activation in human hepatocarcinoma cells. In Vitro Oxidative Stress: GTA-3 Glutathione assay in human hepatocarcinoma cells; GTA-4 Lipid peroxidation assay in human hepatocarcinoma cells; GTA-7 Reactive oxygen species assay in primary hepatocytes. In Vitro Autophagy: GTA-11 Microtubule associated protein light chain 3-I to 3-II conversion by western blot; GTA-12 Autophagic dysfunction in porcine renal proximal tubule cells. In Vitro Drug Release: PHA-1 Radioactive blood partitioning; PHA-2 Ultrafiltration using a stable isotope tracer. In Vivo Pharmacology and Toxicology: NCL performs non–good laboratory practice (GLP) animal studies in rodents to determine absorption, distribution, metabolism, and excretion (ADME) and toxicity profiles. NCL toxicology studies provide identification of target organs of acute and repeat-dose toxicity and may aid in the selection of doses for GLP preclinical and phase 1 human trials. NCL ADME and pharmacokinetic studies track the various components of a nanoparticle formulation in blood and tissues and aid in determination of systemic and tissue exposure, the routes and rates of clearance, and systemic half-life. In vivo ADME-toxicity studies are tailored for each individual nanoparticle.
