Release of FITC-BSA from poly(l-lactic acid) microspheres analysis using flow cytometry

[Display omitted] ► The different sizes of PLA microspheres can be estimated by flow cytometry. ► Flow cytometry provides a fast and exact test to count release of PLA microspheres. ► Fluorescence released from microspheres can be detected by a LAS-3000 image analyzer. In this investigation, biodegr...

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Bibliographic Details
Published in:Colloids and surfaces, B, Biointerfaces B, Biointerfaces, 2012-01, Vol.89 (1), p.271-276
Main Authors: Kuo, Chih-Feng, Tsao, Nina, Chou, Hsin-Hao, Liu, Yi-Ling, Hsieh, Wen-Chuan
Format: Article
Language:English
Subjects:
Animals
aqueous solutions
biodegradability
bovine serum albumin
Cell Line
chloroform
colloids
Degradation
Drugs
emulsifiers
emulsions
Enzymes
evaporation
Flow Cytometry
fluorescein
Fluorescein-5-isothiocyanate - chemistry
glycerol
Glycerols
Homogenizers
Mice
Microsphere
Microspheres
Particle Size
particle size distribution
Phagocytosis
Poly(l-lactic acid) (PLA)
Polyesters - chemistry
Polylactic acid
polyvinyl alcohol
Polyvinyl alcohols
Serum Albumin, Bovine - chemistry
Solvent evaporation
solvents
surfactants
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Summary:[Display omitted] ► The different sizes of PLA microspheres can be estimated by flow cytometry. ► Flow cytometry provides a fast and exact test to count release of PLA microspheres. ► Fluorescence released from microspheres can be detected by a LAS-3000 image analyzer. In this investigation, biodegradable polymer poly(l-lactic acid) (PLA) microspheres were prepared by the W1/O/W2 solvent evaporation method. The inner phase was aqueous solution (W1) that contained bovine serum albumin that was labeled with fluorescein isothiocyanate (FITC-BSA). PLA was dissolved in chloroform with emulsifier sorbitan monooleate (span 80) as the dispersed phase (O). These two solutions (W1/O) were emulsified by a homogenizer to form a primary emulsion. Polyvinyl alcohol (PVA) used as surfactant, was applied in the formation of microspheres (W2). 0.5% (w/v) PLA was stirred at 3000rpm using a homogenizer. Microspheres with sizes of up to around 10μm were produced. These microspheres were separated by the glycerol gradient method, and take microspheres at part of 25% glycerol gradient concentration was analyzed by flow cytometry, indicating a more homogeneous particle size distribution than that not separated. The microspheres were degraded using several enzymes, and around 40% was degraded by 72h. This result reveals the effectiveness of drug delivery by PLA microspheres, which was evaluated by performing a drug release test and flow cytometric analysis. The FITC-BSA concentration in the supernatant increased with the experimental time. At the phagocytosis experiments, encapsulated with FITC-BSA drug of microspheres can be used by the cell, as particle size approximately 1μm.
ISSN:0927-7765
1873-4367
DOI:10.1016/j.colsurfb.2011.09.032