title: Models and methods to evaluate transport of drug delivery systems across cellular barriers.
abstract: Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. Additionally, traversing cellular barriers in the body is crucial for both oral delivery of therapeutic NCs into the circulation and transport from the blood into tissues, where intervention is needed. NC transport across cellular barriers is achieved by: (i) the paracellular route, via transient disruption of the junctions that interlock adjacent cells, or (ii) the transcellular route, where materials are internalized by endocytosis, transported across the cell body, and secreted at the opposite cell surface (transyctosis). Delivery across cellular barriers can be facilitated by coupling therapeutics or their carriers with targeting agents that bind specifically to cell-surface markers involved in transport. Here, we provide methods to measure the extent and mechanism of NC transport across a model cell barrier, which consists of a monolayer of gastrointestinal (GI) epithelial cells grown on a porous membrane located in a transwell insert. Formation of a permeability barrier is confirmed by measuring transepithelial electrical resistance (TEER), transepithelial transport of a control substance, and immunostaining of tight junctions. As an example, ~200 nm polymer NCs are used, which carry a therapeutic cargo and are coated with an antibody that targets a cell-surface determinant. The antibody or therapeutic cargo is labeled with (125)I for radioisotope tracing and labeled NCs are added to the upper chamber over the cell monolayer for varying periods of time. NCs associated to the cells and/or transported to the underlying chamber can be detected. Measurement of free (125)I allows subtraction of the degraded fraction. The paracellular route is assessed by determining potential changes caused by NC transport to the barrier parameters described above. Transcellular transport is determined by addressing the effect of modulating endocytosis and transcytosis pathways.
authors.affiliation: Fischell Department of Bioengineering, University of Maryland.
authors.forename: Rasa
authors.initials: R
authors.lastname: Ghaffarian
authors.forename: Silvia
authors.initials: S
authors.lastname: Muro
mesh_terms: D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D004337:Drug Carriers / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D016503:Drug Delivery Systems / Q000379:methods*; D017097:Electric Impedance; D004847:Epithelial Cells / Q000378:metabolism*; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D007457:Iodine Radioisotopes / Q000737:chemistry; D007553:Isotope Labeling; D053758:Nanoparticles / Q000008:administration & dosage* / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods; D019108:Tight Junctions / Q000378:metabolism; D000519:alpha-Galactosidase / Q000008:administration & dosage / Q000493:pharmacokinetics
chemical_list: D004337:Drug Carriers; D007457:Iodine Radioisotopes; D000519:alpha-Galactosidase