Cell-compatible, multicomponent protein arrays with subcellular feature resolution.

Recent developments in micro/nano-scale technology have enabled the generation of extracellular matrix (ECM) protein microarrays with well-defined geometries. These patterned surfaces have shown utility for the study and control of a variety of cellular behaviors.[1–7] In particular, the patterning of proteins with feature sizes smaller than a single cell have demonstrated potential application for use as tools to control cellular activity.[4, 8] To date, most research has been limited to studies with single protein factors due to technical limitations of existing printing methods. Herein, we describe the development of a microscale direct writing (MDW) technology for the generation of complex ECM protein arrays at subcellular feature size with multiple components. Automated printing techniques based on atomic force microscopy were developed to allow programmable generation of cell-compatible surfaces with multiple ECM proteins, at a subcellular feature size of 6–9 microns. Cell-compatible, two component ECM protein arrays were systematically generated with varying spacing and composition. These arrays were then studied for their effects on cellular attachment and spreading of a model cell line, human myofibroblasts. Interestingly, the precise tuning of spacing and placement two components at subcellular resolution can lead to an increase in cellular alignment. Given the complexity of the in vivo cellular microenvironment, we believe the MDW methods described here could prove generally applicable for the study and optimization of biomaterial surfaces.

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