Plastic Surgery Research Council
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PSRC 60th Annual Meeting
Program and Abstracts

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The Effect Of Bone Marrow Derived Stem Cell Induced By AMD-3100 On Expanded Flap
Hii-Sun Jeong, M.D., Ph.D.1, Hye-Kyung Lee, M.D., Ph.D.2, Kwan-Chul Tark, M.D., Ph.D.3, Dae-Hyun Lew, M.D., Ph.D.3, In Suck Suh, M.D., Ph.D.1.
1Kangnam Sacred Heart Hospital, Hallym University Medical Center, Hallym University College of Medicine, Seoul, Korea, Republic of, 2Department of Plastic and Reconstructive Surgery, Eulji General Hospital, Eulji University School of Medicine, Seoul, Korea, Republic of, 3Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine, Seoul, Korea, Republic of.

Purpose: The purpose of this study is to devise an expanded ischemic flap model, and to investigate the role of AMD-3100 in this model by confirming its effect on mobilization of stem cells from the bone marrow.
Methods: Male Sprague-Dawley rats were used as an animal research model for expanded ischemic skin flaps. First, in a preliminary study, we determined the optimal injection route and dosage of AMD-3100 and investigated the effect of AMD-3100 and silicone sheet insertion between the flap and recipient area. For the main experiments, the rats were divided four groups (n=10 rats per group), including: (1) untreated and not expanded (group I), (2) untreated and expanded (group II), (3) treated with AMD-3100 and not expanded (group III), and (4) treated with AMD-3100 and expanded (group IV). To determine the effects of injection of AMD-3100 on the differentiation and mobilization of hematopoietic stem cells to EPCs, we used flow cytometry using surface markers CD34, VEGFR2, and CD133 and gelatin zymography techniques for active matrix metalloproteinase-9 (MMP-9) in the bone marrow Flap survival and neovascularization were assessed using photometrics, histology (CD31, VEGF, SDF-1, HIF-1α), laser Doppler analysis, western blotting (VEGF, SDF-1, HIF-1α), and real time quantitative RT-PCR (VEGFA, SDF-1, HIF-1α, NOS3, and VEGFR2).
Results:The level of active MMP-9 in the bone marrow was significantly higher in the AMD-3100-treated rats (groups III and IV) than in untreated rats (groups I and II) (p<0.05). The fraction of EPCs (VEGFR2+CD34+, double-positive) in the peripheral blood was significantly greater rats treated with AMD-3100 with expanded skin flaps (group IV) than in any other group at postoperative days 1 and 2 (p<0.05). At the same days, the fraction of single-positive VEGFR2+ cells in the peripheral blood of AMD-3100-treated rats without expanded flaps (group III) was significantly greater than in any other group (p<0.05). The expression of VEGF by immunohistochemistry was significantly increased in groups II, III, and IV in comparison with group I (p<0.05). The expression levels of SDF-1 and VEGFR2 were increased only in group III (p0.05). Treatment with AMD-3100(group III and IV) increased both the number and area of blood vessels (p<0.05).
Conclusion: This endogenous neovascularization induced by AMD-3100 may be a result of the increase in both the area and number of vessels, as well as paracrine augmentation of the expression of VEGF and EPCs. However, the presence of a tissue expander under the flap could block the neovascularization between the flap and the recipient regardless of AMD-3100 treatment and expansion.


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