Comparison of cell interactions with laser machined micron- and nanoscale features in polymer.
Control of cell responses to artificial surfaces is a research goal for much of the biomaterials community. The role that the micron scale topography of a surface can play in controlling cell responses has been well documented and recent advances in nanofabrication techniques have lead to an interest in cells' responses to submicron-scale surface features. The study described here compares the relative influences that nanoscale and micron-scale features exert on cells by examining cytoskeletal organisation. Micron-scale structures were generated on the polyamide Kapton using a 193 nm ArF Excimer laser, at 400 mJ/cm2 fluence. Nanoscale features were generated on Kapton using the excimer laser with a phase mask. Osteoblasts were seeded onto surfaces for 24 h, then the cell membranes were detergent-extracted, and the cells were applied with a primary antibody to actin and a colloidal gold-conjugated secondary antibody. Samples to be examined using the confocal were mounted in glycerol, those for electron microscopy were carbon-coated. The organisation of actin was examined on micron- and nano-scale structures by scoring sections for order of branching and angles of branching to relate changes in the cytoskeleton relative to the control. Although there was a strong influence of micron-scale structures, the cytoskeleton of cells on the nanoscale structures were similar to the controls.
Ball
Michael D
MD
National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland. m.ball@imperial.ac.uk
Prendergast
Una
U
O'Connell
Claire
C
Sherlock
Richard
R
D000199:Actins / Q000276:immunology / Q000378:metabolism; D000906:Antibodies; D002460:Cell Line; D003599:Cytoskeleton / Q000648:ultrastructure*; D006046:Gold; D006801:Humans; D007834:Lasers; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D049329:Nanostructures; D009757:Nylons; D010006:Osteoblasts / Q000648:ultrastructure*
D000199:Actins; D000906:Antibodies; D009757:Nylons; D006046:Gold