Micropatterning Studies of antibodies on PLCL
Date of Issue2012
School of Materials Science and Engineering
Tissue engineering would, more often than not, require cells to be instructed in their growth, so as to align to a specific direction or to differentiate into a specific type of cell. Micropatterning of biomacromolecules onto biocompatible polymers has been a suggested answer to this need. Since most polyesters are hydrophobic and therefore have low cell affinity, surface modification serves as a solution. In this study, spincoated compolymer poly(L-lactide-co ε-caprolactone) (PLCL) film was modified by aminosilane and crosslinked by glutaraldehye for the immobilization of FITC-conjugated gelatin dot and line micropatterns using PDMS. Water contact angle measurements showed increased surface wettability for gelatin-immobilized PLCL films. Immobilization of micropatterns was confirmed by fluorescence microscopy and samples were seeded with human umbilical vein endothelial cells (HUVECs) and cultured for 5 days. HUVECs imaged via inverted microscope showed that the cells were easily directed by line patterns while it took a longer period of time to see the influence of dot patterns. Alamar Blue (AB) assay was done for days 1, 3 and 5. AB fluorescence intensity showed good cell activity on day 1 but subsequent tests showed low cell activity. The HUVECs were fixated and stained for vinculin, F-actin and nucleus for the 3 time points but confocal laser imaging was unable to detect the dyes. These results suggest that PLCL may not be suitable for HUVEC growth.
Final Year Project (FYP)
Nanyang Technological University