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Laminin-Driven Epac/Rap1 Regulation of Epithelial Barriers on Decellularized Matrix

32 Pages Posted: 8 Jul 2019 Publication Status: Accepted

See all articles by Bethany M. Young

Bethany M. Young

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

Keerthana Shankar

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

Cindy K. Tho

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

Amanda R. Pellegrino

Duquesne University - Department of Biomedical Engineering and Nursing

Rebecca L. Heise

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

Abstract

Decellularized tissues offer a unique tool for developing regenerative biomaterials or in vitro platforms for the study of cell-extracellular matrix (ECM) interactions. One main challenge associated with decellularized lung tissue is that ECM components can be stripped away or altered by the detergents used to remove cellular debris. Without characterizing the composition of lung decellularized ECM (dECM) and the cellular response caused by the altered composition, it is difficult to utilize dECM for regeneration and specifically, engineering the complexities of the alveolar-capillary barrier. This study takes steps towards uncovering if dECM must be enhanced with lost ECM proteins to achieve proper epithelial barrier formation. To achieve this, the epithelial barrier function was assessed on dECM coatings with and without the systematic addition of several key basement membrane proteins. After comparing barrier function on collagen, fibronectin, laminin, and dECM in varying combinations as an in vitro coating, the alveolar epithelium exhibited superior barrier function when dECM was supplemented with laminin as evidenced by trans-epithelial electrical resistance (TEER) and permeability assays. Increased barrier resistance with laminin addition was associated with upregulation of Claudin-18, E-cadherin, and junction adhesion molecule (JAM)-A, and stabilization of zonula occludens (ZO)-1 at junction complexes. The Epac/Rap1 pathway was observed to play a role in the ECM-mediated barrier function determined by protein expression and Epac inhibition. These findings reveal potential ECM coatings and molecular therapeutic targets for improved regeneration with decellularized scaffolds or edema related pathologies.

Keywords: Decellularization Lungs Alveolar Laminin Barrier function Epac

Suggested Citation

Young, Bethany M. and Shankar, Keerthana and Tho, Cindy K. and Pellegrino, Amanda R. and Heise, Rebecca L., Laminin-Driven Epac/Rap1 Regulation of Epithelial Barriers on Decellularized Matrix (July 5, 2019). Available at SSRN: https://ssrn.com/abstract=3415223 or http://dx.doi.org/10.2139/ssrn.3415223

Bethany M. Young

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

United States

Keerthana Shankar

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

United States

Cindy K. Tho

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

United States

Amanda R. Pellegrino

Duquesne University - Department of Biomedical Engineering and Nursing

United States

Rebecca L. Heise (Contact Author)

Virginia Commonwealth University (VCU) - Department of Biomedical Engineering

United States

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