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.
Highlights
The development of drugs against pulmonary diseases requires testing of both safety and efficacy
To explore the reproducibility and identify critical factors for aerosol deposition with this device, we investigated the effect of different concentrations, settling times, and multiple depositions (Figure 2)
We describe a new, custom-designed device intended for aerosol deposition into single Transwell R inserts for drug delivery applications of in vitro cell culture models
Summary
The development of drugs against pulmonary diseases requires testing of both safety and efficacy. In this context there recently has been a growing interest in using in vitro cell culture models to replace, reduce, and refine animal experiments (Tannenbaum and Bennett, 2015; Ehrmann et al, 2020). Such tests were and still are performed with submerged cell culture models (Pulskamp et al, 2007; Rothen-Rutishauser et al, 2007; Metz et al, 2020). Modeling physiological situations when developing models and protocols for meaningful in vitro tests is, pivotal (Bastacky et al, 1995; Hiemstra et al, 2018)
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