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Polymers currently utilized for tissue engineering applications do not possess surfaces with nanostructured features. However, the tissue that the polymers will replace is composed of proteins that have nanometer dimensions. Undoubtedly, in situ, cells are accustomed to interacting with surface roughness values in the nanometer regime due to the presence of such proteins in natural tissue. Therefore, the objective of this paper was to design, synthesize and evaluate (using in vitro cellular models) poly-lactic-co-glycolic acid (PLGA) with nanostructured surface features to serve as the next generation of more efficient tissue engineered materials. For this purpose, nanostructured PLGA was created by treating conventional PLGA with various concentrations of NaOH for select periods of time. To eliminate surface chemistry changes created though the etching process, PLGA was cast from silastic molds of NaOH-treated nanostructured PLGA. Results provided the first evidence of increased numbers of vascular cells (specifically, endothelial and aortic smooth muscle cells) and bladder smooth muscle cells on nanostructured compared with conventional PLGA substrates. For this reason, the present results suggest, for the first time, that PLGA should incorporate a high degree of nanostructured surface roughness to enhance tissue regeneration for vascular and bladder applications.