Regulating MC3T3-E1 Cells on Deformable Poly(ε-caprolactone) Honeycomb Films Prepared Using a Surfactant-Free Breath Figure Method in a Water-Miscible Solvent

Honeycomb poly(epsilon-caprolactone) (PCL) films with tunable pore diameters of 3.5, 6.0, and 10 mu m were fabricated directly from solutions in water-miscible, relatively nontoxic tetrahydrofuran using the breath-figure method without assistance of a surfactant. These honeycomb PCL films were characterized in terms of structures and enhanced hydrophobicity. Aiming at fostering bone tissue engineering outcomes, we cultured mouse preosteoblastic MC3T3-E1 cells on these honeycomb films as well as on the flat control, and evaluated their adhesion, spreading, proliferation, alkaline phosphatase (ALP) activity, and calcium content. These cell behaviors were further correlated with the expression levels of integrin subunits of alpha(1), alpha(2), beta(1), and bone-specific gene markers of ALP, collagen type I (COL I), osteocalcin (OCN), and osteopontin (OPN). Honeycomb PCL films remarkably promoted MC3T3-E1 cell adhesion, spreading, proliferation, differentiation, and gene expression. This effect was more prominent when the pore diameter was smaller in the studied range. In addition, honeycomb PCL films were stretched into groove-like structures, on which MC3T3-E1 cells were aligned with a smaller cell area, a higher percentage of aligned cells, and a higher cell elongation ratio when the pores were smaller.