Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery
<b>Background/Objectives</b>: In vitro models play a crucial role in preclinical respiratory research, enabling the testing and screening of mucosal formulations, dosage forms, and inhaled drugs. Mucociliary clearance (MCC) is an essential defense mechanism in mucosal drug delivery but i...
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| author | Janik Martin Veronika Neubauer Rebecca Rittersberger Simon Treitler Patrick Kopp Cemre Günday Iman Shrimo Annabelle Dabbars Frank Rosenau Akif Emre Türeli Nazende Günday-Türeli Oliver Haedicke-Peters Katharina Schindowski |
| author_facet | Janik Martin Veronika Neubauer Rebecca Rittersberger Simon Treitler Patrick Kopp Cemre Günday Iman Shrimo Annabelle Dabbars Frank Rosenau Akif Emre Türeli Nazende Günday-Türeli Oliver Haedicke-Peters Katharina Schindowski |
| author_sort | Janik Martin |
| collection | DOAJ |
| description | <b>Background/Objectives</b>: In vitro models play a crucial role in preclinical respiratory research, enabling the testing and screening of mucosal formulations, dosage forms, and inhaled drugs. Mucociliary clearance (MCC) is an essential defense mechanism in mucosal drug delivery but is often impaired in respiratory diseases. Despite its importance, standardized in vitro MCC assays are rarely reported. Furthermore, many published methods primarily measure cilia beat frequency (CBF), which requires high-speed cameras that are not accessible to all laboratories. Therefore, this study aimed to develop a physiologically relevant, differentiated in vitro model of the respiratory epithelium that incorporates both beating cilia and functional MCC. We chose porcine airway mucosa as an alternative to human tissue due to ethical considerations and limited availability. The established model is designed to provide a reproducible and accessible method for a broad range of research laboratories. <b>Methods</b>: The previously published tracheal mucosal primary cell (TMPC DS) model, derived from porcine tissue, lacked the presence of beating cilia, which are crucial for effective MCC analysis. For accurate MCC assessment, beating cilia are essential as they play a key role in mucus clearance. To address this limitation, the here-described ciliated tracheal mucosal primary cell (cTMPC) model was developed. cTMPCs were isolated from porcine tissue and cultured under air–liquid interface (ALI) conditions for 21 days to promote differentiation. This model was evaluated for cell morphology, tight junction formation, ciliated and mucus-producing cells, barrier function, gene expression, and tracer/IgG transport. MCC and the model’s suitability for standardized MCC assays were assessed using an inverted microscope. In contrast to the TMPC DS model, which lacked beating cilia and thus could not support MCC analysis, the cTMPC model allows for comprehensive MCC studies. <b>Results</b>: The developed differentiated in vitro model demonstrated key structural and functional features of the respiratory epithelium, including well-differentiated cell morphology, tight junction integrity, ciliated and mucus-producing cells, and effective barrier function. Functional MCC was observed, confirming the model’s potential for standardized clearance assays. <b>Conclusions</b>: This differentiated in vitro model closely replicates the structural and functional characteristics of in vivo airways. It provides a valuable platform for studying mucociliary clearance, toxicology, drug uptake, and evaluating mucosal formulations and dosage forms in respiratory research. |
| format | Article |
| id | doaj-art-a66f420f4c1e436f86a5106caaba293c |
| institution | OA Journals |
| issn | 1999-4923 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
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| series | Pharmaceutics |
| spelling | doaj-art-a66f420f4c1e436f86a5106caaba293c2025-08-20T02:28:28ZengMDPI AGPharmaceutics1999-49232025-04-0117446210.3390/pharmaceutics17040462Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug DeliveryJanik Martin0Veronika Neubauer1Rebecca Rittersberger2Simon Treitler3Patrick Kopp4Cemre Günday5Iman Shrimo6Annabelle Dabbars7Frank Rosenau8Akif Emre Türeli9Nazende Günday-Türeli10Oliver Haedicke-Peters11Katharina Schindowski12Institute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyMyBiotech GmbH, Industriestraße 1 B, 66802 Überherrn, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Pharmaceutical Biotechnology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, GermanyMyBiotech GmbH, Industriestraße 1 B, 66802 Überherrn, GermanyMyBiotech GmbH, Industriestraße 1 B, 66802 Überherrn, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, Germany<b>Background/Objectives</b>: In vitro models play a crucial role in preclinical respiratory research, enabling the testing and screening of mucosal formulations, dosage forms, and inhaled drugs. Mucociliary clearance (MCC) is an essential defense mechanism in mucosal drug delivery but is often impaired in respiratory diseases. Despite its importance, standardized in vitro MCC assays are rarely reported. Furthermore, many published methods primarily measure cilia beat frequency (CBF), which requires high-speed cameras that are not accessible to all laboratories. Therefore, this study aimed to develop a physiologically relevant, differentiated in vitro model of the respiratory epithelium that incorporates both beating cilia and functional MCC. We chose porcine airway mucosa as an alternative to human tissue due to ethical considerations and limited availability. The established model is designed to provide a reproducible and accessible method for a broad range of research laboratories. <b>Methods</b>: The previously published tracheal mucosal primary cell (TMPC DS) model, derived from porcine tissue, lacked the presence of beating cilia, which are crucial for effective MCC analysis. For accurate MCC assessment, beating cilia are essential as they play a key role in mucus clearance. To address this limitation, the here-described ciliated tracheal mucosal primary cell (cTMPC) model was developed. cTMPCs were isolated from porcine tissue and cultured under air–liquid interface (ALI) conditions for 21 days to promote differentiation. This model was evaluated for cell morphology, tight junction formation, ciliated and mucus-producing cells, barrier function, gene expression, and tracer/IgG transport. MCC and the model’s suitability for standardized MCC assays were assessed using an inverted microscope. In contrast to the TMPC DS model, which lacked beating cilia and thus could not support MCC analysis, the cTMPC model allows for comprehensive MCC studies. <b>Results</b>: The developed differentiated in vitro model demonstrated key structural and functional features of the respiratory epithelium, including well-differentiated cell morphology, tight junction integrity, ciliated and mucus-producing cells, and effective barrier function. Functional MCC was observed, confirming the model’s potential for standardized clearance assays. <b>Conclusions</b>: This differentiated in vitro model closely replicates the structural and functional characteristics of in vivo airways. It provides a valuable platform for studying mucociliary clearance, toxicology, drug uptake, and evaluating mucosal formulations and dosage forms in respiratory research.https://www.mdpi.com/1999-4923/17/4/462epithelial airway modelinhaled drug deliveryciliabiopharmaceuticsmonoclonal antibodiestherapeutic antibodies |
| spellingShingle | Janik Martin Veronika Neubauer Rebecca Rittersberger Simon Treitler Patrick Kopp Cemre Günday Iman Shrimo Annabelle Dabbars Frank Rosenau Akif Emre Türeli Nazende Günday-Türeli Oliver Haedicke-Peters Katharina Schindowski Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery Pharmaceutics epithelial airway model inhaled drug delivery cilia biopharmaceutics monoclonal antibodies therapeutic antibodies |
| title | Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery |
| title_full | Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery |
| title_fullStr | Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery |
| title_full_unstemmed | Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery |
| title_short | Development and Characterization of a Primary Ciliated Porcine Airway Model for the Evaluation of In Vitro Mucociliary Clearance and Mucosal Drug Delivery |
| title_sort | development and characterization of a primary ciliated porcine airway model for the evaluation of in vitro mucociliary clearance and mucosal drug delivery |
| topic | epithelial airway model inhaled drug delivery cilia biopharmaceutics monoclonal antibodies therapeutic antibodies |
| url | https://www.mdpi.com/1999-4923/17/4/462 |
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