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Research Papers

Biodegradable Nanoparticles Surface Modification Techniques With cIBR Peptide Targeting to LFA-1 Expressing Leukemic Cells

[+] Author and Article Information
Rungsinee Phongpradist

Department of Pharmaceutical Sciences,
Faculty of Pharmacy,
Chiang Mai University,
Chiang Mai 50200, Thailand
e-mail: auan_rx@hotmail.com

Chuda Chittasupho

Department of Pharmaceutical Technology,
Srinakharinwirot University,
Nakornnayok 26120, Thailand

Nutjeera Intasai

Division of Clinical Microscopy,
Department of Medical Technology,
Faculty of Associated Medical Sciences,
Chiang Mai University,
Chiang Mai 50200, Thailand

Teruna J. Siahaan

Professor

Cory J. Berkland

Professor
Department of Pharmaceutical Chemistry,
School of Pharmacy,
The University of Kansas,
Lawrence, KS 66047

Pimlak Charoenkwan

Associate Professor
Department of Pediatrics,
Faculty of Medicine,
Chiang Mai University,
Chiang Mai 50200, Thailand

Songyot Anuchapreeda

Assistant Professor
Division of Clinical Microscopy,
Department of Medical Technology,
Faculty of Associated Medical Sciences,
Chiang Mai University,
Chiang Mai 50200, Thailand

Chadarat Ampasavate

Assistant Professor
Department of Pharmaceutical Sciences,
Faculty of Pharmacy,
Chiang Mai University,
Chiang Mai 50200, Thailand
e-mail: aimchadarat@windowslive.com

1Corresponding author.

Manuscript received July 30, 2012; final manuscript received December 26, 2012; published online March 26, 2013. Assoc. Editor: Henry Hess.

J. Nanotechnol. Eng. Med 3(4), 041005 (Mar 26, 2013) (9 pages) doi:10.1115/1.4023896 History: Received July 30, 2012; Revised December 26, 2012

The lymphocyte function associated antigen-1 (LFA-1) is evaluated for a targeting carrier in leukemia. The cIBR peptide was utilized as the targeting moiety for the drug carrier in direct targeting to LFA-1 expressing cancer cells. This study aims to evaluate the effects of the cIBR peptide conjugation on the specific targeting delivery to the leukemic cell line. Poly (D, L lactide-co-glycolide) (PLGA) nanoparticles were conjugated to the cIBR peptide by three different approaches (coupling, head, and tail) in order to evaluate the nanoparticles' characters, targetability, uptake, drug releasing, and cytotoxicity of each approach. The prepared PLGA nanoparticles were spherical lin shape with a size range of 200–450 nm. The targetability and uptake of three types of cIBR-conjugated nanoparticles (cIBR-NPs) were evidenced and quantified by flow cytometry. The coupling approach presented the highest targetability, uptake, drug releasing, and cytotoxicity followed by the head and tail approaches, respectively. The peptide conjugation method onto the nanoparticles surface was proven to be a key factor for the nanoparticles' physicochemical characteristicss and their efficient delivery.

FIGURES IN THIS ARTICLE
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Copyright © 2013 by ASME
Topics: Nanoparticles , PLGA , Drugs
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References

Figures

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Fig. 1

Scheme of the cIBR-Pluronic-PTX-loaded PLGA-NPs (coupling) preparation

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Fig. 2

Scheme of the (a) head to tail conjugation (head), and (b) tail to head conjugation (tail)

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Fig. 3

cIBR-conjugated PLGA-nanoparticle morphology; (a) cIBR-PEG-NPs (tail), (b) cIBR-PEG-NPs (head), and (c) cIBR-NPs (coupling) by transmission electron microscopy (TEM)

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Fig. 4

The standard curve of paclitaxel for the calculation of the entrapment efficiency

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Fig. 5

The flow cytometry results of the cellular uptake of cIBR-NPs (coupling), cIBR-PEG-NPs (tail), and cIBR-PEG-NPs (head) by Molt-3. Gray-filled histograms represent the fluorescent background of cells, whereas black-filled histograms represent the uptake fluorescent intensity level of three cIBR-conjugated nanoparticles.

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Fig. 6

Kinetic binding profiles of untargeted-NPs, cIBR-PEG-NPs (tail), cIBR-PEG-NPs (head), and cIBR-NPs (coupling) at 5, 30, and 60 min incubations of Molt-3 cells. Data are presented as mean ± S.D. (n = 3), *** indicates p < 0.0001.

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Fig. 7

HPLC chromatogram of free paclitaxel (a) and paclitaxel-loaded nanoparticles separated on a Hypersil ODS 5 μm column (4 × 250 mm i.d.). The mobile phase was a mixture of acetronitrile and water (60:40 (v/v)) delivered at a flow rate of 1.0 ml/min. Detection: 227 nm.

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Fig. 8

In vitro drug release profile of the cIBR-PTX-NPs (coupling), cIBR-PEG-PTX-NPs (tail), and cIBR-PEG-PTX-NPs (head) in a phosphate buffer (pH 7.4) over 30 days at 37 °C. The error bars indicate the standard deviation of mean for n = 3 independent experiments.

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Fig. 9

Cytotoxicity profiles of blank untargeted-nanoparticles and three blank cIBR-conjugated nanoparticles on the Molt-3 cell lines, measured by the MTT assay. Results are represented in triplicate.

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Fig. 10

Cytotoxicity results from free paclitaxel (PTX), cIBR-PTX-NPs, cIBR-PEG-PTX-NPs (head), and cIBR-PEG-PTX-NPs (tail) to the Molt-3 cell lines. Results are represented in triplicate.

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