Contractile smooth muscle cell apoptosis early after saphenous vein grafting Evelio Rodriguez, MD, Erica H Lambert, BS, Michael G Magno, PhD, John D Mannion,

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Contractile smooth muscle cell apoptosis early after saphenous vein grafting Evelio Rodriguez, MD, Erica H Lambert, BS, Michael G Magno, PhD, John D Mannion, MD The Annals of Thoracic Surgery Volume 70, Issue 4, Pages 1145-1152 (October 2000) DOI: 10.1016/S0003-4975(00)01768-9

Fig 1 Desmin stains of a normal saphenous vein (A) and a saphenous vein graft 48 hours after surgery (B). The NSMC bundles (between arrows) are easily differentiated from the SMC bundles (brown cells) (400×). The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 2 Time course of medial SMC and NSMC cell apoptosis. (A) SMC apoptosis at 8 hours was statistically significantly different when compared with time 0 (p < 0.01). (B) NSMC apoptosis at 8, 24, and 48 hours was statistically significantly different when compared with time 0 (p < 0.05). The line represents the mean apoptosis at each time point ± the SEM (bars); open circles represent the apoptotic values for each vein graft. The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 3 Double immunohistochemistry TUNEL/antidesmin of a saphenous vein graft 8 hours after surgery showing multiple SMC (long arrows) and NSMC (short arrows) undergoing apoptosis (800×). The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 4 TUNEL stain of a saphenous vein graft 8 hours after surgery. Apoptotic cells are brown. Multiple SMCs (long arrows) as well as NSMCs (short arrows) are undergoing apoptosis (1,100×). The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 5 Confocal laser scanning microscopy of a saphenous vein 8 hours after grafting. Multiple TUNEL-positive cells (white) demonstrating nuclear condensation, a morphological change characteristic of apoptosis (arrows), within the SMC bundles (light areas) and NSMC bundles (dark band in the middle). The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 6 Electron micrographs depicting SMC at different stages of apoptosis: cell shrinkage (A), nuclear fragmentation (B), and nuclear pyknosis (C). Apoptotic NSMCs were identified showing chromatin condensation (D) and nuclear fragmentation (E) (7,000×). The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 7 Time course of medial cell proliferation. (A) SMC proliferation was minimal. (B) In contrast, NSMCs demonstrated active proliferation. The line represents the mean proliferation at each time point ± the SEM (bars); open circles represent the proliferation values for each vein graft. The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)

Fig 8 PCNA stains of saphenous veins before (A) and 24 hours after grafting (B) revealing multiple NSMCs undergoing proliferation (arrows) while SMC proliferation was a rare finding (800×). The Annals of Thoracic Surgery 2000 70, 1145-1152DOI: (10.1016/S0003-4975(00)01768-9)