title: Cytotoxic effects of pamidronate on monolayers of human intestinal epithelial (Caco-2) cells and its epithelial transport.
abstract: Pamidronate (APD) is a new drug in the treatment of osteolytic bone diseases. Caco-2 cells were used to study the cytotoxic effects of APD on intestinal epithelium and also the transport (mechanism) of APD across the intestinal epithelium. We investigated the cytotoxic effect of APD by combining two spectrophotometric assays [neutral red (NR) uptake and lactate dehydrogenase (LDH) release] with a morphological assay (electron microscopy). The amount of APD transported across the Caco-2 monolayer was measured by HPLC. The permeability of the monolayer was studied by determining the transepithelial electrical resistance (TEER). The results show that after exposing the Caco-2 cells to increasing concentrations of APD [dose range calculated on the basis of relevance to the oral dose administered to patients] the NR uptake decreased while LDH loss increased, which is indicative of a cytotoxic effect of APD. Ultrastructural alterations, including a widening in intercellular spaces and, at higher doses, complete cell death, were observed. The transport percentage of nontoxic doses of APD was low, while the TEER decreased with increasing doses of APD. In conclusion, APD is cytotoxic for Caco-2 cells. As the transport percentage of nontoxic doses of APD is low and APD reduces the TEER, it is hypothesized that APD is transported paracellularly.
authors.affiliation: Department of Clinical Pharmacy, University Hospital Leiden, The Netherlands.
authors.forename: I M
authors.initials: IM
authors.lastname: Twiss
authors.forename: R
authors.initials: R
authors.lastname: de Water
authors.forename: J
authors.initials: J
authors.lastname: den Hartigh
authors.forename: R
authors.initials: R
authors.lastname: Sparidans
authors.forename: W
authors.initials: W
authors.lastname: Ramp-Koopmanschap
authors.forename: H
authors.initials: H
authors.lastname: Brill
authors.forename: M
authors.initials: M
authors.lastname: Wijdeveld
authors.forename: P
authors.initials: P
authors.lastname: Vermeij
mesh_terms: D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002463:Cell Membrane Permeability; D003110:Colonic Neoplasms; D004164:Diphosphonates / Q000493:pharmacokinetics* / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007408:Intestinal Absorption / Q000187:drug effects*; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000187:drug effects*; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D008854:Microscopy, Electron; D008871:Microvilli / Q000187:drug effects / Q000378:metabolism; D008954:Models, Biological; D009499:Neutral Red / Q000493:pharmacokinetics; D000077268:Pamidronate
chemical_list: D004164:Diphosphonates; D009499:Neutral Red; D007770:L-Lactate Dehydrogenase; D000077268:Pamidronate