Poster 1: Cell and Tissue Studies

Apoorva Metkari, Mugdha Pol, Robert Witt (Helen F. Graham Cancer Center), and Xinqiao Jia

University of Delaware

Xerostomia is a salivary gland hypofunction caused by radiation therapy for head and neck cancers. Xerostomia severely affects patients’ quality of life and salivary gland tissue engineering offers a regenerative solution. Previously, we cultured primary human salivary stem/progenitor cells (hS/PCs) in hyaluronic acid (HA)-based matrices with varying proteolytic degradability and demonstrated that cell-mediated matrix degradation is necessary to maintain the pro-acinar stem/progenitor phenotype. However, the multicellular structures developed in these HA gels did not contain correctly polarized lumens. In the current study, we investigate the effects of ROCK inhibition on the phenotypic changes and polarization of hS/PCs. We also study the effects of matrix degradation and ROCK inhibition on stiffness of the cells via mitotracking microrheology. We demonstrate that ROCK inhibition led to the establishment of multicellular structures that were correctly polarized, as evidenced by apical localization of GM130, and basal localization of basement membrane proteins. We also saw an increased expression of acinar markers (AQP3, SLC12A2, AMY1A) at the gene and the protein level in constructs treated with an ROCK inhibitor. In summary, we show that ROCK inhibition promotes the formation of polarized acini structures in HA-based synthetic matrices. This finding is an important step forward in salivary gland tissue engineering.

Research Area: Tissue Engineering