Injury induced neointima formation and its inhibition by retrovirus-mediated transfer of nitride oxide synthase gene in an in-vitro human saphenous vein culture model.
Human saphenous veins were cultured to characterize neointima formation and feasibility of gene transfer to inhibit the intimal proliferative response to injury. Mechanical injury was introduced by abrading the luminal surface of the vein patch with a sterile cotton bud. Both injured and non-injured vein patches were cultured and transduced with retroviral vectors carrying marker or therapeutic genes. After a 14-day culture, the thickness of the intimal layer of non-injured vein patches reached 90+/-28 microm at the edge and 61+/-22 microm at the center (n=29) from the original 22+/-12 microm at harvest (n=6, P=0.02). Mechanical injury to the intimal surface prior to culture resulted in an exaggerated proliferative response. The intimal thickness of injured vein patches increased from 3.4+/-1 microm right after injury to 128+/-23 microm (n=12, P<0.001) at the edge after 14-day culture. Genes were transduced efficiently into a luminal layer of cultured veins using a pseudotyped murine leukemia viral vector. Transduction of gene encoding nitric oxide synthase resulted in reduction of neointima formation to 33+/-7 microm (n=12) at the edge after 14-day culture compared to 90 microm (P<0.01) seen in untransduced non-injured vein patches. Marker gene transduction did not alter intimal proliferative response or its immunohistochemical profile. The data suggest that cultured vein can be used as a model for studying the effects of injury to blood vessels and to evaluate the effects of candidate therapeutic genes.
Yu
Hong
H
Department of Surgery, Vascular Division, Keck School of Medicine of the University of Southern California, 2025 Zonal Avenue, RMR 505, Los Angeles, CA 90089, USA. hongy@hsc.usc.edu
Kumar
S Ram
SR
Tang
Lili
L
Terramani
Thomas T
TT
Rowe
Vincent L
VL
Wang
Ying
Y
Nathwani
Rahul A
RA
Weaver
Fred A
FA
Eton
Darwin
D
D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D006801:Humans; D066298:In Vitro Techniques; D019001:Nitric Oxide Synthase / Q000235:genetics*; D012190:Retroviridae; D012501:Saphenous Vein / Q000201:enzymology / Q000293:injuries; D010959:Tissue Plasminogen Activator / Q000032:analysis; D014170:Transformation, Genetic; D017539:Tunica Intima / Q000201:enzymology* / Q000293:injuries* / Q000473:pathology
D019001:Nitric Oxide Synthase; D010959:Tissue Plasminogen Activator