DNA bound to polypyrrole films: high-resolution imaging, DNA binding kinetics and internal migration.
We have studied the time-dependent uptake of 35S radiolabeled DNA with electrochemically prepared polypyrrole films. The two distinct polypyrrole film surfaces, a rough (solution polymeric growth face, R) and a smooth surface (electrode face, S) were characterized by low-resolution AFM and high-resolution transmission electron microscopy (TEM). These studies showed the presence of steep contours and defects in the form of large and small surface holes and valleys on the rough surface of polypyrrole. The void dimensions ranged from the nanoscale to micron size. By contrast, the smooth surface was flatter and largely devoid of significant structural defects and exhibited closer packing of the polypyrrole chains over large areas. Both surfaces were comprised largely of chains whose average diameters were 1.0-1.2 +/- 0.3 nm. The surface characterization studies were complemented by time-dependent DNA uptake studies which showed a t1/2-dependent total uptake of 35S DNA at higher levels on the rough surface compared to the smooth surface. This is consistent with the apparent higher effective surface area of the rough surface compared to the smooth. Using a proportional counter the time-dependent ratio (R/S) of the 35S DNA detected from the rough surface of the polypyrrole disk to that detected from the smooth surface suggested that DNA was migrating into the disk interior from its uptake surface. The rough side defect dimensions measured by TEM were more than sufficient to allow for the penetration and migration of DNA into the disk interior. Both R/S ratios were extrapolated and found to intersect at an R/S value close to 1.0, suggesting a kinetic process leading ultimately towards a nearly uniform radiolabeled DNA distribution in the disk. These kinetic results were in agreement with the surface characterization studies and suggest a model in which sizeable internal pores exist throughout the electrochemically prepared polypyrrole, that could account for the DNA migration effect. This was confirmed by TEM of the interior of a polypyrrole disk produced by Argon ion milling.
Pande
R
R
Department of Chemistry, Center for Intelligent Biomaterials, University of Massachusetts Lowell, 01854, USA.
Ruben
G C
GC
Lim
J O
JO
Tripathy
S
S
Marx
K A
KA
D000327:Adsorption; D001672:Biocompatible Materials / Q000737:chemistry*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004563:Electrochemistry; D005612:Freeze Drying; D007091:Image Processing, Computer-Assisted; D007700:Kinetics; D018625:Microscopy, Atomic Force; D008854:Microscopy, Electron; D011108:Polymers / Q000737:chemistry; D011758:Pyrroles / Q000737:chemistry*; D013499:Surface Properties
D001672:Biocompatible Materials; D011108:Polymers; D011758:Pyrroles; D004247:DNA