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Dustin M. Bermudez, MD, Junwang Xu, MD, Benjamin J

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1 Inhibition of stromal cell-derived factor-1α further impairs diabetic wound healing 
Dustin M. Bermudez, MD, Junwang Xu, MD, Benjamin J. Herdrich, MD, Antoneta Radu, BS, Marc E. Mitchell, MD, Kenneth W. Liechty, MD  Journal of Vascular Surgery  Volume 53, Issue 3, Pages (March 2011) DOI: /j.jvs Copyright © 2011 Society for Vascular Surgery Terms and Conditions

2 Fig 1 A, Diabetic wounds are shown at day 7 in the green fluorescent protein (GFP)-treated and stromal cell-derived factor inhibitor (SDFi)-treated mice. B, Mean wound area is shown for diabetic mice treated with SDFi or control GFP vector at each day after wounding (each data point, n = 5). C, Results for heterozygote (Het) mice are shown in both experimental groups at day 7. D, Daily mean wound area is shown for heterozygous mice in both groups until closure. The days after wounding at which there was a significant difference in wound size between the two groups is shown by the asterisk above the graph. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Representative sections of wound tissue are shown 7 days after initial wounding (Masson's trichrome stain; black bar = 200 μm): in diabetic wounds treated with (A) lenti-green fluorescent protein (GFP) or (B) with lenti-stromal cell-derived factor-1α inhibitor (SDFi), and heterozygote (Het) wounds treated with (C) GFP or (D) lenti-SDFi. E, The graph shows mean volume of granulation tissue (GT) at day 7 in Db and Het mice treated with GFP and SDFi (error bars show the standard error of the mean). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

4 Fig 3 CD31+ cells are shown at day 7 in a diabetic mice treated with (A) lenti-green fluorescent protein (GFP) or (B) lenti-stromal cell-derived factor-1α inhibitor (SDFi), and in heterozygote (Het) mice treated with (C) GFP or (D) lenti-SDFi (the black bars represent 25 μm). E, The graph shows the mean number of CD31+ cells counted for both groups (the error bars show the standard error of the mean). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

5 Fig 4 α-Smooth muscle actin staining at 7 days after wounding is shown in diabetic mice treated with (A) lenti-green fluorescent protein (GFP) or (B) lenti-stromal cell-derived factor-1α inhibitor (SDFi), and in heterozygote (Het) mice treated with (C) lenti-GFP or with (D) lenti-SDFi (black bar = 25 μm). E, Graph shows the mean number of vessels at day 7 (error bars show the standard error of the mean). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

6 Fig 5 CD45 staining at day 7 is shown in diabetic mice treated with (A) lenti-green fluorescent protein (GFP) or (B) lenti-stromal cell-derived factor-1α inhibitor (SDFi), and in heterozygote (Het) mice treated with (C) lenti-GFP or (D) with lenti-SDFi (black bar = 25 μm). E, Graph shows the mean number of CD45+ cells per high-powered field (HPF; the error bars show the standard error of the mean). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

7 Fig 6 Real-time quantitative polymerase chain reaction analysis of whole wounds at days 3 and 7 after wounding shows (A) interleukin-6 (IL-6) levels in diabetic and nondiabetic mice and (B) macrophage inflammatory protein-2 (MIP-2) levels (n = 5 for each experimental group). GFP, Green fluorescent protein; Het, heterozygote; SDFi, stromal cell-derived factor-1α inhibitor. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

8 Fig 7 Splenic leukocyte migration assay shows there was a decrease in leukocyte migration with increasing concentrations of lenti-stromal cell-derived factor (SDF)-1α inhibitor transfected cells. On average, 256 ± 42 cells were counted in the group with lenti-SDF-1α cells. When the lower chamber had an increased ratio of lenti-SDF-1α inhibitor, fewer cells migrated; when 1.5 × 105 lenti-SDF-1α inhibitor cells were plated, 126 ± 51 cells were counted (*P < .05). The error bars show the standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

9 Fig 8 A, Schematic of the mechanism by which the stromal cell-derived factor (SDF)-1α inhibitor has an effect on the intracellular transduction at the CXCR4 receptor. Although the inhibitor competitively inhibits at CXCR4 through binding, it does not activate a series of important intracellular signaling cascades. B, Inhibition of SDF-1α in diabetic wounds may be easier due to intrinsically low levels. This prevents angiogenesis and subsequently promotes a chronic wound. EPC, Endothelial progenitor cells; HIF-1α, hypoxia inducible factor-1a; IL, interleukin; NFκB, nuclear factor κB. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

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