Patrick C. H. Hsieh, MD, Richard D. Kenagy, PhD, Eileen R

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Presentation transcript:

Bone morphogenetic protein 4: Potential regulator of shear stress-induced graft neointimal atrophy Patrick C.H. Hsieh, MD, Richard D. Kenagy, PhD, Eileen R. Mulvihill, PhD, Joseph P. Jeanette, DO, Xi Wang, MD, PhD, Cindy M.C. Chang, MS, Zizhen Yao, MS, Walter L. Ruzzo, PhD, Suzanne Justice, BS, Kelly L. Hudkins, MS, Charles E. Alpers, MD, Scott Berceli, MD, Alexander W. Clowes, MD Journal of Vascular Surgery Volume 43, Issue 1, Pages 150-158 (January 2006) DOI: 10.1016/j.jvs.2005.08.008 Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 1 Effect of bone morphogenetic proteins (BMP) on smooth muscle cell (SMC) proliferation. A, Dose-dependant effect of BMP2, 4 (B4) and 5 on DNA synthesis in aortic SMCs treated with 10 ng/mL platelet-derived growth factor-BB (P-BB). Data are presented as the percent of PDGF-BB. Diamonds (dashed line), squares (solid line) and triangles (dotted line) represent BMP2, BMP4, and BMP5, respectively. *P ≤ .05, **P ≤ .01, ‡P = .06. B, Dose-dependant effect of BMP4 on DNA synthesis by polytetrafluoroethylene graft neointimal SMCs treated with 10 ng/mL PDGF-BB. Data are presented as the percent of PDGF-BB. **P ≤ .01. C, Reversal by 300 ng/mL noggin (Ng) of the inhibitory effect of 30 ng/mL BMP4 on the stimulation of SMC proliferation by PDGF-BB (10 ng/mL). Data are presented as the percent of the serum-free medium control. **P ≤ .01. Journal of Vascular Surgery 2006 43, 150-158DOI: (10.1016/j.jvs.2005.08.008) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 2 Effects of bone morphogenetic protein 4 (B4) on neointimal smooth muscle cell (SMC) death as determined by enzyme-linked immunoabsorbent assay. A, Effect of 30 ng/mL BMP4 on SMC death after treatment with 10 ng/mL platelet-derived growth factor BB (P-BB) or 0.5% serum for 2 days. *P ≤ .05; **P ≤ .01. B, Time course of SMC death induced by BMP4 (30 ng/mL). Data are presented as percent of the serum-free medium control at each time point. Journal of Vascular Surgery 2006 43, 150-158DOI: (10.1016/j.jvs.2005.08.008) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 3 Expression of bone morphogenetic protein 4 in the polytetrafluoroethylene neointima as determined by Western blot analysis of intimal extracts of grafts exposed to high flow for 4 days (left panel.) A graphic display (right panel) of the combined results from three animals is presented as the percent of normal flow values. Recombinant human (rh) BMP4 was used as a positive control. Arrowheads indicate nonspecific bands as determined by results with non-immune mouse immunoglobulin G. HSS, High shear stress; NSS, normal shear stress; *P ≤ .01. Journal of Vascular Surgery 2006 43, 150-158DOI: (10.1016/j.jvs.2005.08.008) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 4 In situ hybridization for bone morphogenetic protein (BMP4) in the polytetrafluoroethylene (PTFE) intima of grafts exposed to high shear stress for 4 days. A, Antisense sequence of BMP4. The black-silver grains indicate a positive signal. B, Sense sequence of BMP4 for negative control. Arrows indicate the endothelial cells. L, Lumen of PTFE. Magnification ×1000. Journal of Vascular Surgery 2006 43, 150-158DOI: (10.1016/j.jvs.2005.08.008) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 5 Localization of bone morphogenetic protein receptors (BMPRs). Immunostaining (orange/brown) of BMPR-IA (A), -IB (B), and –II (C) in polytetrafluoroethylene (PTFE) grafts exposed to 4 days of normal shear stress. Location of magnified areas (×1000) are indicated by the boxes. L, Lumen of graft; NI, neointima of graft; MA, matrix of PTFE. Magnification ×200. Journal of Vascular Surgery 2006 43, 150-158DOI: (10.1016/j.jvs.2005.08.008) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions