Influence of various cell-detachment solutions on endothelial cells after catheter abrasion for prosthesis colonization prior to implantation.
We (1) evaluated the effectiveness of different media on the detachment of endothelial cells (ECs) from a catheter brush (Cragg thrombolytic catheter system) that was previously used for endothelial cell abrasion in 10 cm human umbilical vein (HUV) segments; (2) tested the practicability of endovascular catheter brush abrasion followed by EC detachment from the catheter brush, in vitro culture of harvested ECs, and finally endothelialization of a prosthesis; and (3) analyzed the defect created by the catheter brush in HUV segments after endovascular catheter abrasion. Best results in detachment of ECs from the catheter brush were obtained with a mixture of phosphate-buffered saline + 1% human albumin. EC vitality was time-dependent in the collected HUV segments postdelivery. Harvested EC viability decreased from (26.28 +/- 5.76)% (0-3 h postdelivery) to (17.29 +/- 4.56)% (after 4-8 h). ECs were easily cultured ex vivo within 2-3 weeks; seeded on nitinol stents, they grew to confluency and formed a monolayer on the stent surface (determined by scanning electron microscopy - SEM). Histological and SEM analysis of HUV segments that had undergone previous catheter brush abrasion revealed slight disruption of the intima but intact subintimal layers. Our findings indicate an advantageous method of capturing and culturing primary ECs for gene therapy or for the analysis and diagnosis of certain blood vessel diseases, especially in cases in which endovascular intervention is performed anyway. Moreover, and of high relevance to the biomaterial field, theoretically the procedure could be used to endothelialze a prosthesis ex vivo for implantation into the patient from whom the ECs were harvested, to reduce the inherent thrombogenicity of the prosthesis.
Priebe
Markus
M
Department of Nuclear Medicine, Ludwig Maximilians University of Munich, LMU, Munich, Germany.
Müller-Hülsbeck
Stefan
S
Jahnke
Thomas
T
Charalambous
Nikolas
N
Hedderich
Jürgen
J
Heller
Martin
M
Paulsen
Friedrich Paul
FP
D017115:Catheter Ablation / Q000379:methods; D002404:Catheterization; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002469:Cell Separation; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D004867:Equipment Design; D019918:Heart Valve Prosthesis Implantation / Q000379:methods; D006801:Humans; D007231:Infant, Newborn; D008855:Microscopy, Electron, Scanning; D015607:Stents; D014471:Umbilical Veins / Q000648:ultrastructure