Polymer substrate topography actively regulates the multicellular organization and liver-specific functions of cultured hepatocytes

This study examines the role of topography of porous synthetic polymer substrates in regulating the tissue-specific morphogenesis of cultured hepatocytes. Porous foams of amorphous 50/50 poly(D,L glycolic-co-lactic acid) (PGLA) with a wide range of controlled pore-size distributions (~1 to 100 μm) were used as culture model surfaces. We found that the induction of microporosity in PGLA substrates significantly improved cell attachment and viability in comparison to those observed on non-porous PGLA films. A detailed evaluation of cellular morphogenesis on the microporous matrices showed that hepatocellular organization was sensitively dependent on the topographical feature size of the foam surfaces. Foams with subcellular size voids (~3 μm) induced kinetics of two-dimensional hepatocyte reorganization, yet limited the extent of three-dimensional aggregation. In contrast, foams with supercellular size voids (~67-μm) restricted hepatocyte motility, thereby promoting the kinetics of 3D aggregation. At intermediate void sizes (~17 μm), both 2D and 3D reorganization kinetics were promoted. Albumin secretory kinetics progressively increased on all void size configurations, the most rapid and sustained kinetics observed in supercellular sized voids, which may serve to minimize cell-polymer contacts and maximize cell-cell contacts in 3D. Overall, these studies demonstrate that void topography of porous polymer substrates is a critical textural feature to induce short-term cell adhesion and viability, and to also selectively regulate the kinetics and extent of multicellular spreading versus 3D aggregation. By virtue of its effects on cell adhesion and morphogenesis, the void topography of nonphysiological polymer scaffolds also is a powerful variable to microengineer hepatospecific activity of tissue analogs.