Influence of micro-well biomimetic topography on intestinal epithelial Caco-2 cell phenotype.
A microfabrication approach was utilized to create topographic analogs of intestinal crypts on a polymer substrate. It was hypothesized that biomimetic crypt-like micro-architecture may induce changes in small intestinal cell (i.e. Caco-2 cell) phenotype. A test pattern of micro-well features with similar dimensions (50, 100, and 500 microm diameter, 50 microm spacing, 120 microm in depth) to the crypt structures found in native basal lamina was produced in the surface of a poly(dimethylsiloxane) (PDMS) substrate. PDMS surfaces were coated with fibronectin, seeded with intestinal-epithelial-cell-like Caco-2 cells, and cultured up to fourteen days. The cells were able to crawl along the steep side walls and migrated from the bottom to the top of the well structures, completely covering the surface by 4-5 days in culture. The topography of the PDMS substrates influenced cell spreading after seeding; cells spread faster and in a more uniform fashion on flat surfaces than on those with micro-well structures, where cell protrusions extending to micro-well side walls was evident. Substrate topography also affected cell metabolic activity and differentiation; cells had higher mitochondrial activity but lower alkaline phosphatase activity at early time points in culture (2-3 days post-seeding) when seeded on micro-well patterned PDMS substrates compared to flat substrates. These results emphasize the importance of topographical design properties of a scaffolds used for tissue engineered intestine.
Wang
Lin
L
Chemical Engineering Department, Northeastern University, 360 Huntington Ave., 342 Snell Engineering, Boston, MA 02115, USA.
Murthy
Shashi K
SK
Fowle
William H
WH
Barabino
Gilda A
GA
Carrier
Rebecca L
RL
D001672:Biocompatible Materials / Q000737:chemistry; D032701:Biomimetics; D018938:Caco-2 Cells / Q000166:cytology / Q000378:metabolism; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002454:Cell Differentiation; D003599:Cytoskeleton / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology; D008422:Materials Testing; D013499:Surface Properties; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods
D001672:Biocompatible Materials