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Facile Dye-Initiated Polymerization of Lactide–Glycolide Generates Highly Fluorescent Poly(lactic-co-glycolic Acid) for Enhanced Characterization of Cellular Delivery
journal contribution
posted on 2020-03-06, 12:38 authored by Mohammad A. Al-Natour, Mohamed D. Yousif, Robert Cavanagh, Amjad Abouselo, Edward A. Apebende, Amir Ghaemmaghami, Dong-Hyun Kim, Jonathan W. Aylott, Vincenzo Taresco, Veeren M. Chauhan, Cameron AlexanderPoly(lactic-co-glycolic acid) (PLGA) is a versatile synthetic copolymer
that is widely used in pharmaceutical applications. This is because
it is well-tolerated in the body, and copolymers of varying physicochemical
properties are readily available via ring-opening polymerization.
However, native PLGA polymers are hard to track as drug delivery carriers
when delivered to subcellular spaces, due to the absence of an easily
accessible “handle” for fluorescent labeling. Here we
show a one-step, scalable, solvent-free, synthetic route to fluorescent
blue (2-aminoanthracene), green (5-aminofluorescein), and red (rhodamine-6G)
PLGA, in which every polymer chain in the sample is fluorescently
labeled. The utility of initiator-labeled PLGA was demonstrated through
the preparation of nanoparticles, capable of therapeutic subcellular
delivery to T-helper-precursor-1 (THP-1) macrophages, a model cell
line for determining in vitro biocompatibility and
particle uptake. Super resolution confocal fluorescence microscopy
imaging showed that dye-initiated PLGA nanoparticles were internalized
to punctate regions and retained bright fluorescence over at least
24 h. In comparison, PLGA nanoparticles with 5-aminofluorescein introduced
by conventional nanoprecipitation/encapsulation showed diffuse and
much lower fluorescence intensity in the same cells and over the same
time periods. The utility of this approach for in vitro drug delivery experiments was demonstrated through the concurrent
imaging of the fluorescent drug doxorubicin (λex =
480 nm, λem = 590 nm) with carrier 5-aminofluorescein
PLGA, also in THP-1 cells, in which the intracellular locations of
the drug and the polymer could be clearly visualized. Finally, the
dye-labeled particles were evaluated in an in vivo model, via delivery to the nematode Caenorhabditis elegans, with bright fluorescence again apparent in the internal tract after
3 h. The results presented in this manuscript highlight the ease of
synthesis of highly fluorescent PLGA, which could be used to augment
tracking of future therapeutics and accelerate in vitro and in vivo characterization of delivery systems
prior to clinical translation.
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nematode Caenorhabditis elegansdye-labeled particlesglycolic aciddye-initiated PLGA nanoparticlespunctate regionssubcellular deliveryTHPFacile Dye-Initiated Polymerizationparticle uptakefluorescence intensitydrug delivery experiments590 nmvivo modelcarrier 5- aminofluorescein PLGArhodamine -6G PLGAring-opening polymerization480 nmPLGA polymersinitiator-labeled PLGAintracellular locationsvivo characterization5- aminofluorescein3 hsubcellular spacesmodel cell lineEnhanced CharacterizationPLGA nanoparticlesphysicochemical propertiesdelivery systemstime periodsdrug delivery carriers24 hpolymer chainfuture therapeuticsSuper resolution confocal fluorescence microscopy imaging
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