Guided cell adhesion, orientation, morphology and differentiation on silicon substrates photolithographically micropatterned with a cell-repellent cross-linked poly(vinyl alcohol) film

In this work, silicon substrates with poly(vinyl alcohol) (PVA) patterns created by a simple, low-cost and high-fidelity photolithographic procedure were evaluated with respect to cell adhesion and alignment, viability, metabolic activity, proliferation and cell cycle progression using the human glioblastoma cell-line U87MG and human skin fibroblasts. In addition, rat adrenal pheochromocytoma cells (PC-12) were employed to evaluate a modified photolithographic protocol appropriate for adhesion of cells requiring extracellular matrix components to adhere on the surface and to demonstrate that the proposed patterned substrates could provide unhindered cell differentiation. Regarding U87MG cells and skin fibroblasts, it was found that as the stripes width increased from 10 to 50 μm, the percentage of cells attached to Si versus the total area (Si + PVA) increased from 78% and 72% to 98.5% and 94.5% (p 95%) to collagen-coated Si stripes when the pattern width was equal to or wider than 10 μm. Following treatment with nerve growth factor, approximately 80% (p < 0.05) of the adherent cells differentiated to neuron-like cells extending neurites exclusively within the pattern. Given that the proposed patterning procedure allows highly selective cell adhesion without affecting cell proliferation, metabolic activity, and differentiation it could serve as a useful tool in various fields including tissue engineering, cell-based sensors and analytical microsystems.