Inkjet printing of laminin gradient to investigate endothelial cellular alignment

To investigate the influence of the protein surface-density gradient on endothelial cell alignment, a novel approach for the fabrication of a laminin gradient on gold-coated substrates has been developed in this study. Our approach involves programmed inkjet printing of an alkanethiol (11-mercaptoundecanoic acid, C10COOH, MUA) gradient onto gold-coated substrates, followed by backfilling with 11-mercapto-1-undecanol (C11OH, MUD). The –COOH moieties were activated and then covalently linked with laminin. This treatment led to a surface-density gradient of laminin. Contact angle measurement, X-ray photoelectron spectroscopy (XPS) and fluorescence microscopy were employed to characterize the self-assembled monolayers (SAMs) and protein gradient, respectively. Results proved the feasibility of the fabrication of a protein gradient by using the inkjet printing technique. The self-assembled monolayer gradients displayed a high packing density, as indicated by dynamic contact angle measurement. More importantly, the gradient slope was easily tunable over a significant distance from 20 to 30 mm. The laminin gradient was clearly visible by fluorescence microscopy observation. Endothelial cells cultured on the surface-density gradient of laminin demonstrated a strong alignment tendency in parallel to the gradient. The higher the laminin density the more cells were observed. The result indicates that cell attachment is dependent on the surface density of laminin. This work broadens our methodology to investigate chemical stimuli-induced cell directional alignment. It is potentially important for understanding cell alignment/ingrowth behavior for angiogenesis and implant technology including tissue-engineered structures.