Substrate architecture effect on siRNA silencing.

• Main Author: Koh, Hwee Ciah.
• Other Authors: Chew Sing Yian
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/16897
• Institution: Nanyang Technological University

Electrospinning of polymeric materials has emerged as a versatile tool for constructing fibrous scaffolds with micro to nanoscale topology similar to the ECM. In this study, human embryonic kidney cells (HEK293) were cultured on electrospun poly (ε-caprolactone) (PCL) fibrous scaffolds. The average diameters of random and aligned fibers were 1.994 ± 0.016 µm and 806 ± 27 nm, respectively. The gene silencing effect in the cells under the influence of topographical cues from the electrospun fibers were evaluated by transfecting HEK 293 cells with siRNAs and TransIT-TKO® reagent. From the real-time PCR results, the difference in fold changes for the aligned and random fibers was found to be not statistically significant (p > 0.05), indicating that fiber alignment does not have a significant impact on gene silencing. To gain a better understanding on the substrate topology effect, more cell lines and more sample size should be involved in the future study.