Effect of retroviral transduction of canine microvascular endothelial cells on beta(1) integrin subunit expression and cell retention to PTFE grafts.
This study evaluated the effect of retroviral transduction on canine microvascular endothelial cell (CMVEC) detachment from fibrin glue coated expanded polytetrafluoroethylene (ePTFE) graft material. CMVEC were isolated from adipose tissue by fluorescent activated cell sorting (FACS). Three treatment groups were evaluated: G-I, transduced CMVEC, selected in antibiotic G418 for 10 days (n=5); G-II, CMVEC selected in G418 and recovered from selection for 4 days (n=5); and G-III, control group of naive CMVEC (n=6). (3)H-thymidine labeled endothelial cells were seeded on fibrin glue coated four-mm diameter PTFE. Grafts were exposed to physiologic shear stresses of 16 dyn/cm(2). Cell detachment was determined by (3)H-thymidine counts in the circuit effluent. beta(1) integrin subunit expression was measured by flow cytometry. After 2 hours of flow exposure, G-I and G-II demonstrated significantly greater cell detachment rates compared with the control seeded grafts. Median peak channel beta(1) integrin subunit value for G-III CMVEC was 2311+/-481.7 vs. 31.5+/-4.51 and 26.3+/-2.0 in the transduced cell groups (p=0.00043). Low beta(1) integrin expression correlated with flow induced high detachment rates of retrovirally-transduced CMVEC.
Grandas
O H
OH
Department of Surgery, Division of Vascular Surgery, Vascular Research Laboratory, University of Tennessee Medical Center at Knoxville, 1924 Alcoa Highway, Box U-11, Knoxville, TN 37920, USA. ograndas@mail.mcg.edu
Costanza
M J
MJ
Donnell
R L
RL
Reddick
T T
TT
Carroll
R C
RC
Stevens
S L
SL
Freeman
M B
MB
Goldman
M H
MH
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001807:Blood Vessel Prosthesis; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D004285:Dogs; D004730:Endothelium, Vascular / Q000166:cytology*; D015718:Fibrin Tissue Adhesive; D005839:Gentamicins / Q000494:pharmacology; D006439:Hemodynamics; D019012:Integrin beta1 / Q000235:genetics* / Q000378:metabolism; D011138:Polytetrafluoroethylene; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic
D000900:Anti-Bacterial Agents; D015718:Fibrin Tissue Adhesive; D005839:Gentamicins; D019012:Integrin beta1; D011138:Polytetrafluoroethylene; C010680:antibiotic G 418