Neutrophil-mediated transport of liposomes across the Madin Darby canine kidney epithelial cell monolayer.
Targeted drug delivery to peripheral blood neutrophils (PMNs) should be of therapeutic potential in various disease states. In addition, substances taken up by PMNs in the circulation may be delivered to an extravascular site via the naturally occurring cell infiltration. The present study employs an in vitro chemotaxis model to test whether particulate drug carriers such as liposomes can be transported across a cellular barrier by migrating PMNs. The system contained 10(7) human PMNs/ml, 0.3-micron liposomes at a total lipid concentration of 2.5 mM, and 10% autologous human serum in the apical side of a confluent Madin Darby canine kidney (MDCK) epithelial cell monolayer of 4.71 cm2. The MDCK cells were grown on a polycarbonate membrane with 3-micron pores without any extracellular matrix, and 10(-7) M f-Met-Leu-Phe was added to the basolateral side as a trigger of chemotaxis. The aqueous phase of the reverse-phase evaporation vesicles (REVs) contained lucifer yellow CH (LY) and [14C]sucrose. The lipid bilayer of the REVs was spiked with [3H]dipalmitoylphosphatidylcholine (DPPC). Transmission electron micrographs showed that, in response to the formyl peptide, PMNs adhered to the apical surface of MDCK cells, emigrated across the MDCK cell layer, passed through the 3-micron pores in the polycarbonate membrane, and finally, appeared in the bottom well. Epifluorescence micrographs showed that most, if not all, of the migrated PMNs contained punctate fluorescence derived from LY. Transport data over a 3.5-hr period indicated that those markers that appeared in the basal side were indeed transported by phagocytosis of REVs by PMNs and that intact serum was an essential component in the process.(ABSTRACT TRUNCATED AT 250 WORDS)
Cho
M J
MJ
Pharmaceutical Research and Development Division, Upjohn Company, Kalamazoo, Michigan 49001.
Scieszka
J F
JF
Cramer
C T
CT
Thompson
D P
DP
Vidmar
T J
TJ
D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D004285:Dogs; D004337:Drug Carriers; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D007546:Isoquinolines / Q000493:pharmacokinetics*; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D008081:Liposomes; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism* / Q000648:ultrastructure; D013395:Sucrose / Q000493:pharmacokinetics*
D004337:Drug Carriers; D007546:Isoquinolines; D008081:Liposomes; D013395:Sucrose; D009240:N-Formylmethionine Leucyl-Phenylalanine; C017475:lucifer yellow