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Nano-Engineered Erythrocyte Ghosts as Inhalational Carriers for Delivery of Fasudil: Preparation and Characterization

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Abstract

Purpose

Nanoerythrosomes (NERs), an engineered derivative of erythrocytes, have long been used as drug delivery carriers. These cell based carriers are biocompatible and biodegradable, and they exhibit efficient drug loading, targeting specificity and prolonged biological half-life. In this study, we have evaluated the feasibility of NERs as inhalable carriers for delivery of fasudil, an investigational drug for the treatment of pulmonary arterial hypertension.

Methods

We prepared NERs by hypotonic lysis of erythrocytes derived from rat blood followed by extrusion through polycarbonate membranes. The formulations were optimized and characterized for size, morphology, entrapment efficiency, stability, cellular uptake and in-vitro release profiles followed by monitoring of drug absorption and safety evaluation after intratracheal administration of fasudil-loaded NERs into rats.

Results

NERs were spherical in shape with an average size of 154.1 ± 1.31 nm and the drug loading efficiency was 48.76 ± 2.18%. Formulations were stable when stored at 4°C for 3 weeks. When incubated with rat pulmonary arterial smooth muscle cells (PASM), a significant amount of NERs was taken up by PASM cells. The drug encapsulated in NERs inhibited the rho-kinase activity upto 50%, which was comparable with the plain fasudil. A ~6–8 fold increase in the half-life of fasudil was observed when encapsulated in NERs.

Conclusion

This study suggests that nanoerythrosomes can be used as cell derived carriers for inhalational delivery of fasudil.

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Abbreviations

ALP:

Alkaline phosphatase

BALF:

Bronchoalveolar lavage fluid

EGM:

Endothelial growth medium

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

HBSS:

Hanks balanced salt solution

HRP:

Horseradish peroxidase

IV:

Intravenous

IT:

Intratracheal

LDH:

Lactate dehydrogenase

LPA:

Lysophosphatidic acid

NERs:

Nanoerythrosomes

PAE:

Pulmonary arterial endothelial

PAH:

Pulmonary arterial hypertension

PASMC:

Pulmonary arterial smooth muscle cells

PBS:

Phosphate buffered saline

ROCK:

Rho-kinase

rMYPT-1:

Recombinant myosin phosphatase target subunit-1

SD:

Sprague Dawley

TMB:

Tetramethylbenzidine

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Acknowledgements and Disclosures

The authors acknowledge Drs. Eva Nozik-Grayck and Kurt Stenmark at the University of Colorado, Denver for providing PASM and PAE cell lines. This work was supported in part by an American Recovery and Reinvestment Act Fund, NIH 1R15HL103431 to Dr. Fakhrul Ahsan.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter Drive, Amarillo, Texas, 79106, USA

    Nilesh Gupta, Brijeshkumar Patel & Fakhrul Ahsan

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  1. Nilesh Gupta
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  2. Brijeshkumar Patel
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  3. Fakhrul Ahsan
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Correspondence to Fakhrul Ahsan.

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Gupta, N., Patel, B. & Ahsan, F. Nano-Engineered Erythrocyte Ghosts as Inhalational Carriers for Delivery of Fasudil: Preparation and Characterization. Pharm Res 31, 1553–1565 (2014). https://doi.org/10.1007/s11095-013-1261-7

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  • DOI: https://doi.org/10.1007/s11095-013-1261-7

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