Internalization of poly(D,L-lactic acid) nanoparticles by isolated human leukocytes and analysis of plasma proteins adsorbed onto the particles.
The objective of this work was to investigate the interactions of poly(D,L-lactic acid) nanoparticles prepared by a recently developed salting-out process, with lymphocytes and monocytes isolated from healthy human donors. Nanoparticles were labeled with a hydrophobic fluorescent dye and incubated with lymphocytes and monocytes, and their uptake was followed by flow cytometry in the presence and absence of plasma. Plasma protein adsorption increased nanoparticle uptake by monocytes, whereas a decrease of cellular binding of the nanoparticles to lymphocytes was noted. The cellular uptake for both cell types consisted in a passive adsorption and in an energy-requiring process, because the cells became 2-3 times more fluorescent when the incubation temperature was increased from 4 to 37 degrees C. When nanoparticles were coated with polyethylene glycol 20,000, uptake by monocytes decreased by 43 and 78% in phosphate-buffered saline and plasma, respectively; a similar decrease in nanoparticle uptake was observed for lymphocytes. Two-dimensional gel electrophoresis was performed to identify the plasma opsonins adsorbed onto the nanoparticle surface. Protein mappings for uncoated and polyethylene glycol-coated nanoparticles differed for two spot series. These spots, not yet clearly identified, may represent specific apolipoproteins involved in the metabolism of human lipoproteins, indicating the possible involvement of specific receptors in the uptake of the nanoparticles.
Leroux
J C
JC
School of Pharmacy, University of Geneva, Switzerland.
Gravel
P
P
Balant
L
L
Volet
B
B
Anner
B M
BM
Allémann
E
E
Doelker
E
E
Gurny
R
R
D000327:Adsorption; D001798:Blood Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D007773:Lactates; D019344:Lactic Acid; D007962:Leukocytes / Q000378:metabolism*; D008863:Microspheres; D009000:Monocytes / Q000378:metabolism; D010078:Oxazines; D011091:Polyesters; D011092:Polyethylene Glycols / Q000494:pharmacology; D011108:Polymers; D014343:Trypan Blue
D001798:Blood Proteins; D005456:Fluorescent Dyes; D007773:Lactates; D010078:Oxazines; D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; D011092:Polyethylene Glycols; C033616:poly(lactide); D014343:Trypan Blue; C044808:nile red