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Treating a collagen scaffold with a low concentration of nicotine promoted angiogenesis and wound healing Pham Hieu Liem, MD, MSc, Naoki Morimoto, MD, PhD, Ran Ito, MD, Katsuya Kawai, MD, PhD, Shigehiko Suzuki, MD, PhD Journal of Surgical Research Volume 182, Issue 2, Pages (June 2013) DOI: /j.jss Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 1 Surgical procedure. (A) Full-thickness skin defects of 8 mm in diameter were created on the backs of mice. (B and C) The collagen sponge was implanted into the defects. (Color version of figure is available online.) Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 2 Macroscopic appearance of the wounds on day 14 after implantation. The wounds were treated with (A) PBS solution, (B) bFGF plus 1.0 × 10−4 M nicotine solution, (C) bFGF solution alone, (D) 1.0 × 10−3 M nicotine solution, (E) 1.0 × 10−4 M nicotine solution, and (F) 1.0 × 10−5 M nicotine solution. The PBS (control) and 1.0 × 10−3 M nicotine groups displayed delayed re-epithelization, whereas the 1.0 × 10−4 M nicotine, bFGF, and bFGF plus 1.0 × 10−4 M nicotine groups displayed a well-advanced wound closure. The wounds in the bFGF plus 1.0 × 10−4 M nicotine group had almost been completely re-epithelized. The diameter of the red circles is 8 mm. (Color version of figure is available online.) Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 3 The epithelized wound area on day 14 after implantation. The epithelized wound area is shown as the mean ± standard error percentage values. The wound area in the 1.0 × 10−3 M nicotine group was significantly larger than that in the other groups (P < 0.01). The wound area in the bFGF plus nicotine group was significantly smaller than that in the 1.0 × 10−4 M nicotine (P < 0.05), control, 1.0 × 10−3 M nicotine, and 1.0 × 10−5 M nicotine groups (P < 0.01). The wound area in the 1.0 × 10−4 M nicotine group was significantly smaller than that in the control, 1.0 × 10−5 M nicotine (P < 0.05), and 1.0 × 10−3 M nicotine groups (P < 0.01). The epithelized wound area in the 1.0 × 10−4 M nicotine group was not significantly different from that in the bFGF alone group (P > 0.05). There was no significant difference between the control and 1.0 × 10−5 M nicotine groups. Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 4 Hematoxylin and eosin staining micrograph of each wound on day 14 after implantation. The wounds were treated with (A) PBS solution, (B) bFGF plus 1.0 × 10−4 M nicotine solution, (C) bFGF alone, (D) 1.0 × 10−3 M nicotine solution, (E) 1.0 × 10−4 M nicotine solution, and (F) 1.0 × 10−5 M nicotine solution. The wounds treated with bFGF plus nicotine were almost completely covered by neoformed epithelia. The vertical black arrows show the wound margins, and the horizontal red arrows indicate the length of the neoformed epithelium (a + b). Scale bar, 500 μm at ×40 magnification. (Color version of figure is available online.) Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 5 Comparison of neoformed epithelium length. Data are presented as the mean ± standard error. The neoformed epithelia in the bFGF plus nicotine group were significantly longer than those in the other groups (P < 0.01). The neoepithelia in the 1.0 × 10−4 M nicotine group were longer than those in the control (P < 0.01) and 1.0 × 10−5 M nicotine solution groups (P < 0.05). The neoepithelia in the 1.0 × 10−3 M nicotine solution group was significantly shorter than those in the other groups (P < 0.05 versus control, P < 0.01 versus bFGF, 1.0 × 10−4 M nicotine, and bFGF plus nicotine groups). The neoepithelium length of the bFGF alone group was not significantly different from that of the 1.0 × 10−4 M nicotine group (P > 0.05). There was no significant difference between the control and 1.0 × 10−5 M nicotine groups. Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 6 Immunohistochemical staining of neoformed capillaries above the muscle layer using the vWF antibody. The wounds were treated with (A) PBS solution, (B) bFGF plus 1.0 × 10−4 M nicotine solution, (C) bFGF solution alone, (D) 1.0 × 10−3 M nicotine solution, (E) 1.0 × 10−4 M nicotine solution, and (F) 1.0 × 10−5 M nicotine solution. On day 14 after implantation, neoformed capillaries were detected in all experimental groups, and the number of capillaries was greatest in the bFGF plus nicotine group. The red arrowheads indicate capillaries. Scale bar, 100 μm at ×200 magnification. (Color version of figure is available online.) Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 7 Quantitative analysis of capillary area. Data are presented as the mean ± standard error. The capillary area in the 1.0 × 10−4 M nicotine group was larger than that in the control, 1.0 × 10−5 M nicotine solution (P < 0.05), and 1.0 × 10−3 M nicotine solution groups (P < 0.01). The stained area in the PBS (control) group was significantly smaller than that in the bFGF (P < 0.01) and bFGF plus 1.0 × 10−4 M nicotine groups (P < 0.01). There was no significant difference between the control and 1.0 × 10−5 M nicotine solution groups. Journal of Surgical Research , DOI: ( /j.jss ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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