An Atelocollagen Coating for Efficient Local Gene Silencing by Using Small Interfering RNA.
In the last decades, many efforts have been made to counteract adverse effects after stenting atherosclerotic coronary arteries. A breakthrough in better vascular wall regeneration was noted in the new era of drug-eluting stents. A novel personalized approach is the development of gene-eluting stents promising an alteration in gene expression involved in regeneration. We investigated a coating system consisting of the polymer atelocollagen (ATCOL) and a specific small interfering RNA (siRNA) for intercellular adhesion molecule-1 (ICAM-1) found on the surface of defective endothelial cells (ECs). We demonstrated very high cell viability, in which EA.hy926 grew on 0.008% or 0.032% ATCOL layers. Additionally, hemocompatibility assays proved the biocompatibility of this coating. The highest transfection efficiency with EA.hy926 was achieved with 5 μg siRNA immobilized in ATCOL after 2 days. The release of fluorescent-labeled siRNA was about 9 days. Long-term knockdown of ICAM-1 was analyzed by flow cytometry, revealing that the coating with 0.008% ATCOL and 5 μg siICAM-1 provoked gene silencing up to 8 days. 5'-RNA ligase-mediated rapid amplification of cDNA ends PCR (RLM-RACE-PCR) demonstrated the specificity of our established ATCOL gene-silencing coating, meaning that our coating is well suited for further investigations in in vivo studies. Herein, we would like to demonstrate that our ATCOL is well-suited for better artery wall regeneration after stent implantation.
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Olivia
O
Koenig
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Dimitrios
D
Nothdurft
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Nadja
N
Perle
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Bernd
B
Neumann
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Andreas
A
Behring
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Ilka
I
Degenkolbe
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Tobias
T
Walker
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Christian
C
Schlensak
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany. Electronic address: hans-peter.wendel@med.uni-tuebingen.de.
Hans Peter
HP
Wendel
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Tuebingen, 72076 Baden-Wuerttemberg, Germany.
Andrea
A
Nolte
atelocollagen; atherosclerosis; gene knockdown; local delivery; siICAM-1; siRNA transfection