ABSTRACT
The in vitro endothelial response of primary bovine aortic endothelial cells (BAECs) was investigated on a flat Ti surface vs a nanostructured TiO2 nanotube surface. The nanotopography provided nanoscale cues that facilitated cellular probing, cell sensing, and especially cell migration, where more organized actin cytoskeletal filaments formed lamellipodia and locomotive morphologies. Motile cell protrusions were able to probe down into the nanotube pores for contact stimulation, and focal adhesions were formed and disassembled readily for enhanced advancement of cellular fronts, which was not observed on a flat substrate of titanium. NOx and endothelin-1 functional assays confirmed that the nanotubes also up-regulated an antithrombic cellular state for maintaining vascular tone. The enhanced endothelial response to TiO2 nanotubes is significant for a potential modification of vascular stent surfaces in order to increase the rate and reliability of endothelialization and endothelial cell migration onto the stent for repairing arterial injury after activation.
Subject(s)
Cell Movement , Endothelial Cells/cytology , Endothelial Cells/metabolism , Nanotubes/chemistry , Titanium/metabolism , Actins/metabolism , Animals , Cattle , Cell Shape , Cells, Cultured , Endothelin-1/metabolism , Microscopy, Electron, Scanning , Nanotubes/ultrastructure , Nitric Oxide/metabolism , Surface Properties , Vinculin/metabolismABSTRACT
We describe here a new in vitro protocol for structuring cardiac cell cultures to mimic important aspects of the in vivo ventricular myocardial phenotype by controlling the location and mechanical environment of cultured cells. Microlithography is used to engineer microstructured silicon metal wafers. Those are used to fabricate either microgrooved silicone membranes or silicone molds for microfluidic application of extracellular matrix proteins onto elastic membranes (involving flow control at micrometer resolution). The physically or microfluidically structured membranes serve as a cell culture growth substrate that supports cell alignment and allows the application of stretch. The latter is achieved with a stretching device that can deliver isotropic or anisotropic stretch. Neonatal ventricular cardiomyocytes, grown on these micropatterned membranes, develop an in vivo-like morphology with regular sarcomeric patterns. The entire process from fabrication of the micropatterned silicon metal wafers to casting of silicone molds, microfluidic patterning and cell isolation and seeding takes approximately 7 days.
