Electrospun fiber-mediated siRNA delivery and Gene silencing

While substrate topography influences cell responses, RNA interference (RNAi) has also emerged as a potent method for understanding and directing cell fate. In particular, the three dimensional (3D) electrospun fibers can offer biomimicking topographical signal to alter cell phenotypes, while RNAi can guide cell behaviors through regulating gene expression. Herein, we aim to establish platforms to understand and enhance cellular internalization of small interfering RNA (siRNA) and gene silencing by electrospun nanofiber-mediated RNAi. First, we examined the influence of fiber architecture on siRNA-mediated gene silencing in human somatic cells. The model cell, human dermal fibroblasts (HDFs), were cultured onto aligned and randomly oriented electrospun poly(-caprolactone) (PCL) fibers of different average diameters (300 nm, 700 nm and 1.3 m). It is observed that decreasing fiber diameter from 1.3 m to 300 nm, regardless of fiber orientation, significantly improved housekeeping gene Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and low abundance gene Collagen I silencing efficiencies by ~ 3.8 and ~ 4.4 folds respectively (p