Authors

Justus C. Horstmann,1,2 Chelsea R. Thorn,3 Patrick Carius,1,2 Florian Graef,1,† Xabier Murgia,1,*† Cristiane de Souza Carvalho-Wodarz,1,* and Claus-Michael Lehr1,2

Affiliations

1Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Saarbrücken, Germany
2Department of Pharmacy, Saarland University, Saarbrücken, Germany
3Clinical and Health Science, University of South Australia, Adelaide, SA, Australia
Edited by: Nihal Engin Vrana, Sparta Medical, France
Reviewed by: Eder Lilia Romero, National University of Quilmes, Argentina; Anja Lena Thiebes, RWTH Aachen University, Germany; Catherine Fromen, University of Delaware, United States

Topic

Cell Biology, Medical Research

Abstract

The deposition of pre-metered doses (i.e., defined before and not after exposition) at the air–liquid interface of viable pulmonary epithelial cells remains an important but challenging task for developing aerosol medicines. While some devices allow quantification of the deposited dose after or during the experiment, e.g., gravimetrically, there is still no generally accepted way to deposit small pre-metered doses of aerosolized drugs or pharmaceutical formulations, e.g., nanomedicines. Here, we describe a straightforward custom-made device, allowing connection to commercially available nebulizers with standard cell culture plates. Designed to tightly fit into the approximately 12-mm opening of either a 12-well Transwell® insert or a single 24-well plate, a defined dose of an aerosolized liquid can be directly deposited precisely and reproducibly (4.8% deviation) at the air–liquid interface (ALI) of pulmonary cell cultures. The deposited dose can be controlled by the volume of the nebulized solution, which may vary in a range from 20 to 200 μl. The entire nebulization-deposition maneuver is completed after 30 s and is spatially homogenous. After phosphate-buffered saline (PBS) deposition, the viability and barrier properties transepithelial electrical resistance (TEER) of human bronchial epithelial Calu-3 cells were not negatively affected. Straightforward in manufacture and use, the device enables reproducible deposition of metered doses of aerosolized drugs to study the interactions with pulmonary cell cultures grown at ALI conditions.

DOI: https://dx.doi.org/10.3389%2Ffbioe.2021.643491

Extract

An inverted fluorescent microscope (Olympus IX53) connected to CoolLed pE-300 illuminator system was used with a 2 × objective to visualize the deposition of sodium fluorescein-LCNPs on the membranes, from the bottom side up.

Product Associated Features

Stable and precise illumination with the pE-300 Series enables quantitative analysis, which is vital for measuring the performance of the novel device developed in this research.

Product Type

pE-300white, pE-300lite, pE-300ultra

Journal

Front. Bioeng. Biotechnol.

Year of Publication

2021

Country of Publication

Australia, France, Germany