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Arterial heparan sulfate proteoglycans inhibit vascular smooth muscle cell proliferation and phenotype change in vitro and neointimal formation in vivo 

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Presentation on theme: "Arterial heparan sulfate proteoglycans inhibit vascular smooth muscle cell proliferation and phenotype change in vitro and neointimal formation in vivo "— Presentation transcript:

1 Arterial heparan sulfate proteoglycans inhibit vascular smooth muscle cell proliferation and phenotype change in vitro and neointimal formation in vivo  John A. Bingley, MMedSci, Ian P. Hayward, PhD, Julie H. Campbell, PhD, Gordon R. Campbell, PhD  Journal of Vascular Surgery  Volume 28, Issue 2, Pages (August 1998) DOI: /S (98) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

2 Fig. 1 Typical elution profile for digested arterial tissue from DEAE-cellulose anion exchange chromatography column. Fractions of 4 mL (monitored by absorbance at 280 nm) were collected at a flow rate of 45 mL/hr with a 0.2 to 1.5 mol/L saline gradient. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

3 Fig. 2 The 4% to 15% polyacrylamide gel showing the three bands of HSPGs (arrows) within the first peak eluted from anion exchange chromatography. Samples of 250 ng were added to each lane. Lane 1, untreated sample from peak 1 (see Fig. 1). Lane 2, peak 1 after heparitinase action. Molecular weights are indicated (right) (Alcian blue-silver stain). The band at approximately 70 kDa (lane 2) has bovine serum albumin used as stabilizer in the heparitinase preparation used to degrade the heparan sulfate GAG. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

4 Fig. 3 Intima-to-media ratios 14 days after de-endothelialization in rabbit carotid arteries and periadventitial treatment of the distal section of artery with 25% pluronic gel containing saline (control) (n = 8), 2000 μg of Enoxaparin (n = 8), or 680 μg of HSPG (n = 9). Values are mean ± SEM. *Significantly different at P < .05. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

5 Fig. 4 Representative light microscopic sections showing the effects of topical delivery on the degree of neointimal formation in rabbit distal common carotid arteries at 14 days after de-endothelialization. a, Control, gel only. b, Enoxaparin (2000 μg). c, HSPG (680 μg). L = lumen; IEL = internal elastic lamina; M = media; N = neointima. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

6 Fig. 5 In vitro release of HSPG from 25% F-127 pluronic gel showing the amount of 35S-labeled HSPG remaining in a 25% F-127 pluronic gel at 37°C after removal of overlying PBS at various time points. A line-of-best-fit shows a half-life of approximately 30 hours. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

7 Fig. 6 Mean volume fraction of myofilaments (±SEM) of primary rabbit aortic SMCs cultured in Medium 199 plus 5% fetal calf serum in the presence of 34 μg/mL HSPG, 100 μg/mL Enoxaparin, or medium only (control) for 5 days. The proportion of cytoplasm filled with myofilaments (%Vvmyo), indicative of SMC phenotype, was calculated by point-counting morphometry. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

8 Fig. 7 Representative electron micrographs of primary rabbit aortic SMCs cultured for 5 days in Medium 199 plus 5% fetal calf serum with (a) medium only, (b) 100 μg/mL Enoxaparin, and (c) 34 μg/mL HSPG. Myo = myofilament; E.R. = endoplasmic reticulum; M = mitochondria. Original magnification, 12,500×. Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

9 Fig. 8 3H-Thymidine incorporation by (A) rabbit aortic SMCs in Medium 199 plus 5% fetal calf serum at day 6 in primary culture and (B) third-passage rabbit aortic SMCs in Medium 199 plus 5% fetal calf serum at day 3 after treatment with HSPGs (●), Enoxaparin (■), or chondroitin sulfate proteoglycans (▵) at varying doses, shown as a percentage of control. Values are mean ± SEM of three separate measurements (eight replicates per experiment). Journal of Vascular Surgery  , DOI: ( /S (98) ) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

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