TY - JOUR
T1 - Platelet-derived growth factor production by cells from Dacron grafts implanted in a canine model
AU - Pitsch, R. J.
AU - Minion, D. J.
AU - Goman, M. L.
AU - Van Aalst, J. A.
AU - Fox, P. L.
AU - Graham, L. M.
PY - 1997
Y1 - 1997
N2 - Purpose: Previous studies of grafts implanted in dogs documented a time- dependent increase in platelet-derived growth factor (PDGF) production that correlated with inner-capsule thickness. The purpose of this study was to identify the cells in vascular grafts that produce PDGF. Methods: Dacron thoracoabdominal grafts were seeded with autologous endothelial cells (ECs), implanted in 11 beagles, and removed after 4 or 20 weeks. ECs and smooth muscle cells (SMCs) were cultured from grafts and adjacent aorta, and PDGF in the conditioned media was measured by radioreceptor assay. The PDGF A-chain mRNA level in freshly harvested cells was assessed using reverse transcriptase, followed by polymerase chain reaction, and expressed as a ratio of glyceraldehyde-3-phosphate dehydrogenase signal. Localization of PDGF A-chain and B-chain protein was also examined with immunohistochemical analysis. Results: Graft and aortic ECs in primary culture did not produce significantly different amounts of PDGF in 72 hours, averaging 368 ± 160 and 340 ± 81 pg/μg DNA, respectively. Graft SMCs in primary culture produced significantly more PDGF than aortic SMCs (584 ± 343 and 113 ± 94 pg/μg DNA, respectively; p < 0.01). Graft SMC PDGF secretion remained greater than aortic SMC PDGF secretion through at least six cell passages. PDGF A-chain mRNA levels were not significantly different for aortic or graft ECs. The PDGF A-chain mRNA level was significantly higher for graft SMCs than aortic SMCs (2.44 ± 0.67 and 1.45 ± 0.57 pg/μg, respectively; p < 0.03). Immunocytochemical analysis detected PDGF A-chain and B-chain protein in the ECs from both native aorta and graft as well as the subendothelial SMCs in the graft, but not in the SMCs of the native aorta. Conclusions: These results suggest that graft SMCs are functionally altered, producing more PDGF than aortic SMCs. PDGF produced by graft SMCs may contribute to the development of intimal hyperplasia.
AB - Purpose: Previous studies of grafts implanted in dogs documented a time- dependent increase in platelet-derived growth factor (PDGF) production that correlated with inner-capsule thickness. The purpose of this study was to identify the cells in vascular grafts that produce PDGF. Methods: Dacron thoracoabdominal grafts were seeded with autologous endothelial cells (ECs), implanted in 11 beagles, and removed after 4 or 20 weeks. ECs and smooth muscle cells (SMCs) were cultured from grafts and adjacent aorta, and PDGF in the conditioned media was measured by radioreceptor assay. The PDGF A-chain mRNA level in freshly harvested cells was assessed using reverse transcriptase, followed by polymerase chain reaction, and expressed as a ratio of glyceraldehyde-3-phosphate dehydrogenase signal. Localization of PDGF A-chain and B-chain protein was also examined with immunohistochemical analysis. Results: Graft and aortic ECs in primary culture did not produce significantly different amounts of PDGF in 72 hours, averaging 368 ± 160 and 340 ± 81 pg/μg DNA, respectively. Graft SMCs in primary culture produced significantly more PDGF than aortic SMCs (584 ± 343 and 113 ± 94 pg/μg DNA, respectively; p < 0.01). Graft SMC PDGF secretion remained greater than aortic SMC PDGF secretion through at least six cell passages. PDGF A-chain mRNA levels were not significantly different for aortic or graft ECs. The PDGF A-chain mRNA level was significantly higher for graft SMCs than aortic SMCs (2.44 ± 0.67 and 1.45 ± 0.57 pg/μg, respectively; p < 0.03). Immunocytochemical analysis detected PDGF A-chain and B-chain protein in the ECs from both native aorta and graft as well as the subendothelial SMCs in the graft, but not in the SMCs of the native aorta. Conclusions: These results suggest that graft SMCs are functionally altered, producing more PDGF than aortic SMCs. PDGF produced by graft SMCs may contribute to the development of intimal hyperplasia.
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U2 - 10.1016/S0741-5214(97)70149-6
DO - 10.1016/S0741-5214(97)70149-6
M3 - Article
C2 - 9240324
AN - SCOPUS:0030855275
SN - 0741-5214
VL - 26
SP - 70
EP - 78
JO - Journal of Vascular Surgery
JF - Journal of Vascular Surgery
IS - 1
ER -