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Biological Effects of Simvastatin Formulated as pMDI on Pulmonary Epithelial Cells

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ABSTRACT

Purpose

The aim of this study is to evaluate the biological effects of Calu-3 epithelial cells in response to the delivery of simvastatin (SV) via solution pressurized metered dose inhaler (pMDI).

Methods

SV pMDI was aerosolised onto Calu-3 air-interface epithelial cells using a modified glass twin stage impinger. The transport of SV across Calu-3 cells, mucus production, inflammatory cytokines production i.e., interleukin (IL) 6, 8 and tumour necrosis factor alpha (TNF- α) and oxidative stress from Calu-3 cells following treatment with SV pMDI was investigated and compared to untreated cells.

Results

It was found that SV had the ability to penetrate into the respiratory epithelium and convert into its active SV hydroxy acid (SVA) metabolite. Furthermore, the amount of mucus produced was significantly reduced when SV was deposited on Calu-3 compared to untreated cells. Additionally, SV delivered by pMDI reduces production of IL-6, 8 and TNF-α from Calu-3 following stimulation with lipopolysaccharide (LPS). SV also showed equivalent antioxidant property to vitamin E.

Conclusions

Treatment with SV solution pMDI formulation on Calu-3 cells reduces mucus production, inflammatory cytokines and oxidative stress. This formulation could potentially be used clinically as muco-inhibitory and anti-inflammatory therapy for treatment of chronic lung diseases.

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Abbreviations

%:

Percentage

AIC:

Air interface culture

ATCC:

American type culture collection

COPD:

Chronic obstructive pulmonary disease

Da:

Dalton

DCF:

Dichlorofluorescin

DMEM:

Dulbecco’s modified eagle’s medium

DMSO:

Dimethyl sulfoxide

ELISA:

Enzyme-linked immunosorbent assay

EtOH:

Ethanol

Flu-Na:

Fluorescein sodium

h:

Hour

HBSS:

Hank’s balanced salt solution

HFA:

Hydrofluoroalkane

HFA-134a:

1,1,1 2-tetrafluoroethane

HMG-CoA:

Hydroxy-3-methyl-glutaryl-Coenzyme

HPLC:

High-performance liquid chromatography

IL:

Interleukin

L:

Litre

L/min:

Litre per minutes

LPS:

Lipopolysaccharide

mg:

Milligram

mL:

Millilitres

mM:

Millimolar

mm:

Millimetre

MW :

Molecular weight

NADPH:

Nicotinamide adenine dinucleotide phosphate

NF-κB:

Nuclear factor kappa

PBS:

Phosphate buffer solution

pMDI:

Pressurised metered dose inhaler

RGB:

Red-green-blue

ROS:

Reactive oxygen species

s:

Second

SD:

Standard deviation

SV:

Simvastatin

SVA:

Simvastatin hydroxy acid

TEER:

Transepithelial electrical resistance

TNF-a:

Tumour necrosis factor alpha

TSI:

Twin stage impinger

v/v :

Volume per volume

w/w :

Weight per weight

μg:

Microgram

μg/mL:

Microgram per millilitre

μL:

Microliter

μm:

Micrometer

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ACKNOWLEDGMENTS AND DISCLOSURES

Professor Young is the recipient of an Australian Research Council Future Fellowship (project number FT110100996). A/Professor Traini is the recipient of an Australian Research Council Future Fellowship (project number FT12010063).

Alaa Tulbah is also grateful to Umm Al-Qura University for the scholarship and Saudi Government for their financial support.

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Authors and Affiliations

  1. Respiratory Technology, Woolcock Institute of Medical Research and Discipline of Pharmacology, Sydney Medical School, Sydney University, Sydney, NSW, 2037, Australia

    Alaa S. Tulbah, Hui Xin Ong, Wing-Hin Lee, Paul M. Young & Daniela Traini

  2. Faculty of Pharmacy, Umm Al Qura University, Makkah, Saudi Arabia

    Alaa S. Tulbah

  3. Department of Pharmacy, University of Parma, Parma, Italy

    Paolo Colombo

Authors
  1. Alaa S. Tulbah
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  2. Hui Xin Ong
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  6. Daniela Traini
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Correspondence to Daniela Traini.

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Tulbah, A.S., Ong, H.X., Lee, WH. et al. Biological Effects of Simvastatin Formulated as pMDI on Pulmonary Epithelial Cells. Pharm Res 33, 92–101 (2016). https://doi.org/10.1007/s11095-015-1766-3

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