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Distinct Patterns of Angiogenesis in Oral and Skin Wounds
A.M. Szpaderska1,
C.G. Walsh1,
M.J. Steinberg2 and
L.A. DiPietro1,*
1 Burn and Shock Trauma Institute, and
2 Division of Oral and Maxillofacial Surgery, Department of Surgery, Loyola University Medical Center, 2160 S. First Ave., Maywood, IL 60153;
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Figure 1. Time course of angiogenesis in oral mucosal and skin wounds. The vascular density of the wounds on days 1, 3, 5, and 7 after injury was determined by image analysis of CD31-stained tissues. Data are expressed as mean ± SE (n = 5). Scale bars (lower right corner of each panel): 200 µm. (A) Immunohistochemical localization of vessels in the wound bed was performed with anti-CD31 antibody. The photographs depict immunostained histologic sections of wounds from days 5 and 7 after wounding occurred. Original magnification x50. (B) Vessel density in normal skin and wound bed. The % vascularization [(CD31-positive area/Total wound bed area) x 100] was determined for 2 wound sections from each of 5 mice. (C) Increase in vessel density in wound bed over the baseline (normal tissue vasculature). We calculated the change in vascularity by dividing the vasculature value of each day by baseline value. Data are expressed as mean ± SE (n = 5). p < 0.001, mucosa vs. skin. NS, normal skin.
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Figure 2. Levels of pro-angiogenic factors in oral mucosal and skin wounds. Wound tissue was collected and homogenized, and the supernatant was analyzed by ELISA. (A) VEGF (n = 4). p < 0.001, mucosa vs. skin. (B) FGF-2 (n = 4).
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Figure 3. VEGF mRNA expression in oral and epidermal keratinocytes. (A) To determine relative changes in mRNA levels, we compared the densitometry values for 493-bp PCR product (VEGF165) with β-actin expression and normalized them by setting the highest value to 100. The results are depicted graphically as the mean ± SE (n = 3). *p < 0.05 hypoxia vs. normoxia. (B) A representative gel of RT-PCR performed on RNA isolated from keratinocytes grown under normoxic and hypoxic conditions. N = normoxia, H = hypoxia.
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Figure 4. VEGF production by oral and epidermal keratinocytes. 1 x 105 oral and skin keratinocytes were incubated under hypoxia for 18 hrs. Culture supernatants were collected and analyzed (by ELISA) for VEGF release. As a control, VEGF protein concentrations were determined in media of cells grown under normoxia. The bars represent the mean ± SE (n = 3). **p < 0.01 hypoxia vs. normoxia.
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Journal of Dental Research, Vol. 84, No. 4,
309-314 (2005)
DOI: 10.1177/154405910508400403
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