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Efficacy of Piroxicam Plus Cisplatin-Loaded PLGA Nanoparticles in Inducing Apoptosis in Mesothelioma Cells

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Abstract

Purpose

Combined treatment based on cisplatin-loaded Poly(D,L-lactic-co-glicolic)acid (PLGA) nanoparticles (NP-C) plus the NSAID piroxicam was used as novel treatment for mesothelioma to reduce side effects related to cisplatin toxicity.

Methods

PLGA nanoparticles were prepared by double emulsion solvent evaporation method. Particle size, drug release profile and in vitro cellular uptake were characterized by TEM, DLS, LC/MS and fluorescence microscopy. MSTO-211H cell line was used to analyse NP-C biological efficacy by FACS and protein analysis.

Results

Cisplatin was encapsulated in 197 nm PLGA nanoparticles with 8.2% drug loading efficiency and 47% encapsulation efficiency. Cisplatin delivery from nanoparticles reaches 80% of total encapsulated drug in 14 days following a triphasic trend. PLGA nanoparticles in MSTO-211H cells were localized in the perinuclear space NP-C in combination with piroxicam induced apoptosis using a final cisplatin concentration 1.75 fold less than free drug. Delivered cisplatin cooperated with piroxicam in modulating cell cycle regulators as caspase-3, p53 and p21.

Conclusions

Cisplatin loaded PLGA nanoparticles plus piroxicam showed a good efficacy in exerting cytotoxic activity and inducing the same molecular apoptotic effects of the free drugs. Sustained cisplatin release allowed to use less amount of drug, decreasing toxic side effects. This novel approach could represent a new strategy for mesothelioma treatment.

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Abbreviations

ACN:

Acetonitrile

C:

Cisplatin

C/P:

Cisplatin plus piroxicam

CE:

Collision energy

COX:

Cyclooxygenase

CTRL:

Untreated cells

DDTC:

Diethyldithiocarbamate

DLS:

Dynamic light scattering

DMF:

N,N-dimethylformamide

DP:

Declustering potential

DPH:

1,6-Diphenyl-1,3,5-hexatriene

ECL:

Enhanced ChemiLuminescence

EE:

Encapsulation efficiency

EP:

Entrance potential

FACS:

Fluorescence activated cell sorting

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

HPLC:

High performance liquid chromatography

i :

Cisplatin release phases

k i :

Release constant value for each release phase

LC/MS:

Liquid chromatography/mass spectrometry

LE:

Loading efficiency

NP:

PLGA-nanoparticles

NP-C:

Cisplatin-loaded PLGA nanoparticles

NP-C/P:

Cisplatin-loaded nanoparticles plus piroxicam

NP-F:

Fluorescent PLGA nanoparticles

NSAID:

Non-steroidal anti-inflammatory drug

PBS:

Phosphate buffered saline

PDI:

Poly dispersion index

PI:

Propidium iodide

PLGA:

Poly(D,L-lactic-co-glicolic)acid

Pt-(DDTC)2 :

Cisplatin-DDTC derivative

PVA:

Polyvinyl alcohol

PVDF:

Polyvinylidene fluoride

RCC1:

Regulator of chromosome condensation 1

RIPA:

Radio-immunoprecipitation assay buffer

RPMI-1640:

Roswell Park Memorial Institute culture medium

SDS-PAGE:

Sodium dodecyl sulphate - polyacrylamide gel electrophoresis

SIM:

Selected ion monitoring

TEM:

Transmission electron microscopy

TOC:

Total organic carbon

w/o:

Water/oil

w/o/w:

water/oil/water

y i (0):

Cisplatin concentration at the beginning of each release phase

y i (t):

Cisplatin amount released over the time

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ACKNOWLEDGMENTS AND DISCLOSURES

We thank INBB for supporting Dr. Maria Teresa Piccolo and Dr. Ciro Menale with fellowships; Dr. Laura Pisapia for helping in FACS analysis; Dr. Rosarita Tatè for helping in microscopy analysis; the FACS and the IM IGB Facilities. We thank Prof. Maria Rosaria Chiummo for help in the revision of the manuscript. We also thank the Neapolitan section of the IIT, in particular Dr. RaffaeleVecchione, for using Zetasizer.

Author information

Authors and Affiliations

  1. Department of Experimental Medicine, Second University of Naples, Naples, Italy

    Ciro Menale, Carla Nicolucci & Nadia Diano

  2. Gene Expression & Molecular Genetics Laboratory, Institute of Biosciences and Bioresources, CNR, via P. Castellino, 111-80131, Naples, Italy

    Ciro Menale, Maria Teresa Piccolo, Ilaria Favicchia, Maria Grazia Aruta & Stefania Crispi

  3. National Institute of Biostructures and Biosystems, Viale Medaglie d’Oro, 305-00136, Rome, Italy

    Ciro Menale, Maria Teresa Piccolo, Alfonso Baldi, Damiano Gustavo Mita, Stefania Crispi & Nadia Diano

  4. Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Naples, Italy

    Maria Teresa Piccolo & Alfonso Baldi

  5. Department of Experimental Medicine, University of Messina, Messina, Italy

    Carla Nicolucci

  6. Biophysics Laboratory, Institute of Genetics and Biophysics, CNR, Naples, Italy

    Vincenzo Barba, Damiano Gustavo Mita & Nadia Diano

Authors
  1. Ciro Menale
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  2. Maria Teresa Piccolo
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  4. Maria Grazia Aruta
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  8. Damiano Gustavo Mita
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Corresponding authors

Correspondence to Damiano Gustavo Mita or Stefania Crispi.

Additional information

Ciro Menale and Maria Teresa Piccolo equally contributed to the work.

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Menale, C., Piccolo, M.T., Favicchia, I. et al. Efficacy of Piroxicam Plus Cisplatin-Loaded PLGA Nanoparticles in Inducing Apoptosis in Mesothelioma Cells. Pharm Res 32, 362–374 (2015). https://doi.org/10.1007/s11095-014-1467-3

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  • DOI: https://doi.org/10.1007/s11095-014-1467-3

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