The proliferative response to platelet-derived growth factor of smooth muscle cells isolated from synthetic vascular grafts in a canine model David J.

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The proliferative response to platelet-derived growth factor of smooth muscle cells isolated from synthetic vascular grafts in a canine model David J. Minion, MD, Maarten-Paul van de Kerkhove, MD, Greg R. Goodman, MD, John A. van Aalst, MD, Afaf Absood, PhD, Paul L. Fox, PhD, Linda M. Graham, MD Journal of Vascular Surgery Volume 29, Issue 5, Pages 845-851 (May 1999) DOI: 10.1016/S0741-5214(99)70212-0 Copyright © 1999 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig 1 Growth of graft and aortic smooth muscle cells (SMCs) in primary culture. Equal numbers of SMCs harvested from aortas and Dacron grafts were plated in M199 medium (Sigma Chemical Company, St Louis, Mo) and 20% fetal bovine serum. Cell counts were performed after 1, 2, 4, 6, and 8 days. SMCs from five dogs were studied, and representative growth curve expressed as log2 of the cell count at time x/cell count at baseline is depicted. Journal of Vascular Surgery 1999 29, 845-851DOI: (10.1016/S0741-5214(99)70212-0) Copyright © 1999 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig 2 Uptake of [3H]thymidine in response to platelet-derived growth factor (PDGF). Early passage graft and aortic smooth muscle cells were plated at a density of 20,000 cells/cm 2 in M199 medium (Sigma Chemical Company, St Louis, Mo) that contained 20% fetal bovine serum (FBS) and were allowed to attach overnight. Smooth muscle cells were made quiescent in medium with FBS 0.5% for 48 hours, and then were incubated in DME/F12 (Irvine Scientific, Santa Ana, Calif) that contained 0.5% FBS and varying concentrations of PDGF-AB. Proliferative response, as determined with [3H]thymidine uptake during hours 18 and 24 of the incubation, is depicted. In each segment, there was an increase in PDGF (10 ng/mL) group as compared with control (P < .05, with Student paired t test). Relative increase was decreased in graft as compared with proximal and distal aortas (P < .05, with analysis of variance). Journal of Vascular Surgery 1999 29, 845-851DOI: (10.1016/S0741-5214(99)70212-0) Copyright © 1999 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig 3 Cell count in response to platelet-derived growth factor (PDGF). Early passage graft and aortic smooth muscle cells were plated overnight in M199 medium (Sigma Chemical Company, St Louis, Mo) with 20% fetal bovine serum and then were made quiescent for 48 hours. Smooth muscle cells then were incubated in DME/F12 (Irvine Scientific, Santa Ana, Calif) that contained 0.5% fetal bovine serum and varying concentrations of PDGF (0, 0.1, 1, and 10 ng/mL). Cell number was determined at baseline (beginning of incubation) and after 72 hours of exposure to medium with and without PDGF. In each segment, there was an increase in the PDGF (10 ng/mL) group as compared with control (P < .05, with Student paired t test). Relative increase was decreased in graft as compared with proximal and distal aortas (P < .05, with analysis of variance). Journal of Vascular Surgery 1999 29, 845-851DOI: (10.1016/S0741-5214(99)70212-0) Copyright © 1999 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig 4 Response to platelet-derived growth factor (10 ng/mL) as function of duration of graft implantation. Data from proximal and distal aortas are combined. Note that at each time point relative increase in graft smooth muscle cells was less than that of aortic smooth muscle cells. Journal of Vascular Surgery 1999 29, 845-851DOI: (10.1016/S0741-5214(99)70212-0) Copyright © 1999 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig 5 Binding characteristics of platelet-derived growth factor (PDGF)–BB. With 24-well plates, passage one smooth muscle cells were plated at density of 50,000 cells/cm2 in M199 medium (Sigma Chemical Company, St Louis, Mo) that contained fetal bovine serum 20% and then were made quiescent in M199 medium with fetal bovine serum 0.5% for 48 hours. Cell counts were performed, and PDGF receptor binding then was determined with 125I-labeled PDGF-BB. Maximal binding (femtomole/million cells) was decreased in graft as compared with proximal and distal aortas (P < .05, with analysis of variance). Dissociation constant (picomolar) did not differ significantly. Journal of Vascular Surgery 1999 29, 845-851DOI: (10.1016/S0741-5214(99)70212-0) Copyright © 1999 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions