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Subacute limb ischemia induces skeletal muscle injury in genetically susceptible mice independent of vascular density  Joseph M. McClung, PhD, Timothy.

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Presentation on theme: "Subacute limb ischemia induces skeletal muscle injury in genetically susceptible mice independent of vascular density  Joseph M. McClung, PhD, Timothy."— Presentation transcript:

1 Subacute limb ischemia induces skeletal muscle injury in genetically susceptible mice independent of vascular density  Joseph M. McClung, PhD, Timothy J. McCord, BS, Kevin Southerland, MD, Cameron A. Schmidt, BS, Michael E. Padgett, BS, Terence E. Ryan, PhD, Christopher D. Kontos, MD  Journal of Vascular Surgery  Volume 64, Issue 4, Pages e2 (October 2016) DOI: /j.jvs Copyright © 2016 Society for Vascular Surgery Terms and Conditions

2 Fig 1 Tissue pathology after subacute femoral artery occlusion with two ameroid constrictors (ACs). Acute limb ischemia (ALI) or subacute femoral artery occlusion using 2 ACs was performed in BL6 and BALB/c mice, and animals were monitored for 7 days (ALI, n = 16 per strain; 2AC, n = 8 BL6, n = 13 BALB/c). A, Representative image and schematic of sites of ligation/resection for ALI or 2AC placement on the femoral artery. The arrows indicate sites of constrictor placement or resection: (1) Lateral circumflex femoral; (2) Proximal caudal femoral; (3) Superficial caudal epigastric artery; (4) Saphenous artery; (5) Popliteal artery. B, Ex vivo image of an AC at day (d) 7, demonstrating complete occlusion. C, Distribution of limb necrosis scores during the week of ischemia (d1, d3, d5, d7) induced by ALI in BL6 and BALB/c mice. *P < .05 vs day-matched BL6. D, Distribution of limb necrosis scores during the week of ischemia (d1, d3, d5, d7) induced by 2AC in BL6 and BALB/c mice. *P < .05 vs day-matched BL6. E, Representative hematoxylin and eosin (H&E) sections demonstrating tibialis anterior (TA) morphology 7 days after ALI and 2AC surgery. L, Left (surgical limb); R, right (nonsurgical limb). Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Comparison of limb blood flow between BALB/c and BL6 mice after either acute limb ischemia (ALI) or subacute femoral artery occlusion with two ameroid constrictors (ACs). A, Representative images from laser Doppler perfusion imaging in BALB/c and BL6 mice before and after ALI or 2AC placement up to 7 days (d) postoperatively. B, Quantitation of paw perfusion, represented as a ratio of ischemic (L) to nonischemic (R) limb perfusion (ALI, n = 10 per strain; 2AC, n = 8 per strain). *P < .05 vs day- and ischemic-model-matched BL6. Post, Postoperative; Pre, preoperative. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

4 Fig 3 Comparison of BALB/c and BL6 pathology and blood flow after further refinement of subacute femoral artery occlusion by placement of a single ameroid constrictor (1AC). A, Representative image of a BALB/c mouse after placement of an AC on the proximal portion of the femoral artery, immediately proximal to the epigastric arterial branch. The arrow indicates site of constrictor placement: (1) Femoral artery; (2) Lateral circumflex femoral; (3) Superficial caudal epigastric artery. B, Representative gross anatomy photos and photomicrographs of tibialis anterior (TA) muscle from BALB/c and BL6 mice subjected to subacute femoral artery occlusion via the placement of a single AC on the proximal femoral artery (1AC) for 15 days (d). C, Representative images from laser Doppler perfusion imaging in BALB/c and BL6 mice before and after 1AC placement up to 28 days postoperatively. D, Quantitation of paw perfusion in BALB/c (n = 29) and BL6 (n = 27) mice, represented as a ratio of ischemic (L) to nonischemic (R) limb perfusion. *P < .05 vs day- and ischemic-model-matched BL6. Post, Postoperative; Pre, preoperative. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Skeletal muscle pathology and macrophage infiltration after single ameroid constrictor (1AC)-mediated subacute ischemia. A, Representative hematoxylin and eosin (H&E) sections demonstrating tibialis anterior (TA) morphology and immunostaining of macrophages (white arrows) after 7 days (d) of 1AC subacute ischemia. L, Left (surgical limb); R, right (nonsurgical limb). B, Quantitation of the percentage total TA area immunostaining positive for macrophages (% TA Area Mac1+). *P < .05 vs strain-specific contralateral (R) control. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

6 Fig 5 Genetic divergence of ischemic skeletal muscle myofiber morphology after single ameroid constrictor (1AC)-mediated subacute ischemia. BALB/c and BL6 mice were subjected to subacute femoral artery occlusion via the placement of a single AC on the proximal femoral artery (1AC) for 15 days (d). A, Representative hematoxylin and eosin (H&E) sections demonstrating tibialis anterior (TA) morphology after 15 days of 1AC subacute ischemia, L, Left (surgical limb; gray bar); R, right (nonsurgical limb; black bar). B, Percentage TA myofibers with centralized nuclei. C, Percentage TA noncontractile tissue (NCT). Values are presented as means ± standard error of the mean (SEM). *P < .05 vs strain-specific contralateral (R) control. D, Myofiber cross-sectional area (CSA) in BALB/c and BL6 TA. Mean CSAs are presented as means ± standard error of the mean (SEM) % change from contralateral (R) control. *P < .05 vs BL6. Distribution of skeletal muscle myofiber size was calculated in (E) BALB/c and (F) BL6 TA muscles. Values are presented as means ± SEM. *P < .05 vs strain-specific contralateral (R) control. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

7 Fig 6 Skeletal muscle vascular density after single ameroid constrictor (1AC)-induced subacute limb ischemia (SLI). A, Tibialis anterior (TA) muscle sections from BALB/c and BL6 mice 15 days (d) after 1AC-induced SLI were stained as indicated for the visualization of myofibers (dystrophin), capillaries/endothelial cells (CD31), intermediate vessels (SMA), and nuclei (DAPI). Representative sections are shown. B, TA CD31+ capillary vessel density. C, TA SMA+ vessel density. D, Average TA SMA+ vessel area. Values are presented as means ± standard error of the mean (SEM). L, Left (surgical limb; gray); R, right (nonsurgical limb; black). Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

8 Fig 7 Ischemic skeletal muscle regeneration after sustained single ameroid constrictor (1AC)-mediated subacute ischemia. A, Representative hematoxylin and eosin (H&E)-stained sections demonstrating tibialis anterior (TA) morphology 28 days (d) after 1AC placement. Sections of TA muscle were analyzed for mean myofiber cross-sectional area (CSA) at day 28 and presented as a percentage of the right, nonsurgical limb (B). *P < .05 vs BL6. C, The percentages of total myofibers with centralized nuclei. *P < .05 vs strain-specific contralateral (R) control. All values are presented as means ± standard error of the mean (SEM). Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

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