||The role of burn blister fluid in burn wound neovascularization
||Institute of Clinical Medicine
Burn blister fluids
Burn wound healing is a complex and dynamic process, which involves an interaction between different cells and mediators. This process can be divided into inﬂammatory phase, proliferative phase, and remodeling phase. Neovascularization, beginning during the inﬂammatory phase of wound healing, is an imperative stage of wound healing. The whole process consists of not only angiogenesis but also adult vasculogenesis.
Superﬁcial partial thickness burn (SPTB) heals within 2 weeks without scarring. Deep partial thickness burn (DPTB), on the other hand, heals beyond 2 weeks and requires aggressive treatment to prevent hypertrophic scarring. Burn blisters on the skin are a hallmark of not only SPTB but also DPTB. Previous study reported that blisters contained several cytokines with beneﬁcial effects on keratinocyte proliferation. However, the effect of burn blister ﬂuids on the neovascularization has not been fully explored. Burn hypertrophy scar usually exhibits in a hypervascular status. DPTB wounds often result in hypertrophy scar, we assume that the neovascularization between SPTB and DPTB wounds may be different.
To verify this hypothesis, neovasculogenic effect of two different kinds of burn blister ﬂuids was tested and compared in vitro and in vivo. The results show DPTB blister ﬂuids more predominantly support neovascularization than do SPTB wounds in the early stage of wound healing.
Following the above study, we are particularly interested in investigating the factors modulating the differential angiogenic activity of these fluids. An expression analysis of angiogenic factors in burn fluids was conducted. Upon finding a significant difference of angiogenin between the two different burn fluids, the role of angiogenin was assessed in vitro, in vivo and human study. We find that angiogenin clearly involves in the neovascularization of burn wound healing.
Rapid and accurate assessment of burn depth is the most important determinant of burn wound management in the early stage of burn injury. However, assessment of second-degree burn wounds with intact blisters has always been difficult, even for experienced surgeons. Although our finding demonstrates a significantly higher angiogenin expression in DPTB blister fluids than formed in SPTB fluids, screening the angiogenin in burn blister fluids for determination of burn depth should be further studied. Nevertheless, application of burn fluid contents to observe cell-mediators interaction in burn wounds is of considerable interest to those investigating the processes of burn wound healing.
CONTENT LIST PAGE
Chinese abstract 3
English abstract 5
Chapter 1 Burn Blister in Burn Wound Healing
1.1 Phases of Normal Wound Healing 15
1.1.1 Inflammation phase 15
1.1.2 Proliferative phase 16
188.8.131.52 Angiogenesis 17
184.108.40.206 Vasculogenesis 18
220.127.116.11.1 Characterization of circulating EPC 19
18.104.22.168.2 Role of EPCs in Neovascularization 20
22.214.171.124.3 Mechanisms by which EPCs promote 21
126.96.36.199.4 Mobilization of EPCs 22
188.8.131.52.5 Chemotaxis of EPCs 22
184.108.40.206.6 Homing and differentiation of EPCs 23
220.127.116.11.7 Vasculogenesis in wound healing 24
1.1.3 Remodeling phase 26
1.2 The Burn Wound 27
1.2.1 Anatomy of normal skin 27
1.2.2 Thermal effect on skin 28
1.2.3 Depth of the Burn 29
1.2.4 First-degree burn 29
1.2.5 Second-degree burn 29
1.2.6 Third-degree burn 30
1.2.7 Fourth-degree burn 30
1.2.8 Assessment of burn wound depth 30
18.104.22.168 Clinical evaluation 31
22.214.171.124 Biopsy and histology 32
126.96.36.199 Thermography 33
188.8.131.52 Vital dyes 34
184.108.40.206 Indocyanine green video angiography 35
220.127.116.11 Laser Doppler flowmetry 35
18.104.22.168 Nuclear imaging 37
22.214.171.124 Noncontact and high-frequency 38
1.3 Burn blister 39
1.3.1 Occurrence of blister 39
1.3.2 Advantages of blisters intact 40
1.3.3 Advantages of blister removal 41
1.4 Conclusion 43
1.5 Goal of Ph.D project 44
Chapter 2 The Effect of Superficial And Deep Partial Thickness Burn Blister Fluid in Burn Wound Neovascularization
2.1 Introduction 46
2.2 Materials and Methods 46
2.2.1. Patient samples 46
2.2.2 Immunohistochemistry 47
2.2.3 Cell proliferation assay 48
2.2.4 Isolation and differentiation of circulating 48
2.2.5 Isolation of CD34+ and CD14+ cells 49
2.2.6 Transwell migration assay 49
2.2.7 Real-time polymerase chain reaction (PCR) 50
2.2.8 Characterization of endothelial cells 50
2.2.9 Matrigel plug assay 51
2.2.10 Statistical analysis 51
2.3 Results 51
2.3.1 Microvascular density in different burn 51
2.3.2 Proliferative ability of endothelial cells in 52
burn blister ﬂuids
2.3.3 Migration ability of CACs in burn blister 52
2.3.4 Endothelial gene expression in burn blister 52
2.3.5 Production of functional endothelial cells in 53
burn blister fluids
2.3.6 In-vivo animal study of burn blister fluids 54
2.3.7 Purity of CD34+ and CD14+ cells 54
2.3.8 Migration ability of CD34+ and CD14+ cells 54
in burn blister fluids
2.3.9 The endothelial gene expression of CD34+ 55
and CD14+ cells in burn blister fluids
2.3.10 Endothelial characteristics of CD34+ and 55
CD14+ cells in burn blister fluids
2.4 Discussion 55
2.5 Conclusion 58
Chapter 3 Factors modulating the angiogenic activity of burn blister fluids
3.1 Introduction 60
3.2 Materials and Methods 60
3.2.1. Patient samples 60
3.2.2 Cytokine antibody assay 61
3.2.3 Enzyme-linked immunosorbent assay (ELISA) 61
3.2.4 Cell proliferation assay 62
3.2.5 CAC isolation and differentiation 62
3.2.6 Endothelial cell characterization 62
3.2.7 Matrigel plug assay 63
3.2.8 Immunohistochemistry 63
3.2.9 Statistical analysis 64
3.3 Results 64
3.3.1 Differential angiogenesis-related cytokine 64
expression in burn blister fluids
3.3.2 Effect of endothelial cell proliferation by 65
3.3.3 Correlation of angiogenin expression with 66
3.3.4 CACs differentiation by angiogenin inhibition 66
3.3.5 Angiogenin neutralization attenuated 67
neovascularization in vivo
3.3.6 Angiogenin expression in human burn wound 67
3.4 Discussion 68
3.5 Conclusion 72
Chapter 4 Conclusion 73
Chapter 5 Future Work 75
Reference List 76
List of thesis related publications 122
Curriculum Vitae 123
LIST of TABLES
Table 1 The number of CAC differentiation in patients 95
according to burn severity
Table 2 ELISA analysis of cytokines in blister fluids 96
Table 3 Angiogenin staining intensity in human burn 97
LIST of FIGURES
Figure 1. Common methods of “EPC” culture 98
Figure 2. Origin and differentiation of endothelial 99
Figure 3. Immunohistochemical staining of CD31 in 100
SPTB and DPTB wounds
Figure 4. The effect of burn ﬂuids on endothelial 101
Figure 5. The migration ability of peripheral blood 102
mononuclear cells induced by SPTB or DPTB
Figure 6. The effect of burn blister ﬂuids on the 103
endothelial mRNA expression
Figure 7A. The effects of burn blister fluids on CACs 104
Figure 7B. Flow cytometry analysis of endothelial 105
differentiation of CACs
Figure 8. In vivo angiogenic activity of burn 106
Figure 9. Purity of CD34+ and CD14+ cells 107
Figure 10. Migration ability of CD34+ and CD14+ 108
cells in DPTB blister fluid
Figure 11. The vWF mRNA gene expression of cultured 109
CD34+ and CD14+ cells in DPTB blister
Figure 12A.Flow cytometry analysis of CACs 110
differentiation from cultured CD34+,
CD34-, CD14+ and CD14- cells.
Figure 12B.Differentiation of cultured CD34+ and 111
CD14+ cells in DPTB blister fluids
Figure 13. Differential expression of cytokines in 112
SPTB and DPTB fluids
Figure 14. Time course of cytokines expression in 113
burn blister fluid following thermal
Figure 15. The effect of anti-angiogenin antibodies 114
on endothelial cell proliferation
Figure 16. The effect of different levels of 115
anti-angiogenin antibody on endothelial
Figure 17. The effect of angiogenin on CAC 116
Figure 18. The effect of neutralizing antibody to 117
angiogenin on CAC differentiation induced
by DPTB fluid
Figure 19. The effect of VEGF-A neutralization on 118
DPTB fluid-induced CAC differentiation
Figure 20. Angiogenin neutralization in DPTB fluids 119
reduced neovascularization in Matrigel plugs
Figure 21. The relationship between angiogenin and 120
neovascularization in human burn wounds
Figure 22. Summary of differential neovascularization 121
between superficial and deep partial
1. Singer, A.J. & Clark, R.A. Cutaneous wound healing. N Engl J Med 341, 738-746 (1999).
2. Gottrup, F. Oxygen in wound healing and infection. World J Surg 28, 312-315 (2004).
3. Duval, C., et al. Proliferation and wound healing of vascular cells trigger the generation of extracellular reactive oxygen species and LDL oxidation. Free Radic Biol Med 35, 1589-1598 (2003).
4. Engelhardt, E., et al. Chemokines IL-8, GROalpha, MCP-1, IP-10, and Mig are sequentially and differentially expressed during phase-specific infiltration of leukocyte subsets in human wound healing. Am J Pathol 153, 1849-1860 (1998).
5. Amento, E.P. & Beck, L.S. TGF-beta and wound healing. Ciba Found Symp 157, 115-123; discussion 123-119 (1991).
6. Leibovich, S.J. & Ross, R. The role of the macrophage in wound repair. A study with hydrocortisone and antimacrophage serum. Am J Pathol 78, 71-100 (1975).
7. Ross, R., Everett, N.B. & Tyler, R. Wound healing and collagen formation. VI. The origin of the wound fibroblast studied in parabiosis. J Cell Biol 44, 645-654 (1970).
8. Krueger, W.W., et al. Fibroblast implantation enhances wound healing as indicated by breaking strength determinations. Otolaryngology 86, ORL-804-811 (1978).
9. Clark, R.A., Nielsen, L.D., Welch, M.P. & McPherson, J.M. Collagen matrices attenuate the collagen-synthetic response of cultured fibroblasts to TGF-beta. J Cell Sci 108 ( Pt 3), 1251-1261 (1995).
10. DiPietro, L.A. Wound healing: the role of the macrophage and other immune cells. Shock 4, 233-240 (1995).
11. Mellin, T.N., et al. Acidic fibroblast growth factor accelerates dermal wound healing. Growth Factors 7, 1-14 (1992).
12. Carmeliet, P. Mechanisms of angiogenesis and arteriogenesis. Nat Med 6, 389-395 (2000).
13. Martinez, J., Ferber, A., Bach, T.L. & Yaen, C.H. Interaction of fibrin with VE-cadherin. Ann N Y Acad Sci 936, 386-405 (2001).
14. Schimmenti, L.A., Yan, H.C., Madri, J.A. & Albelda, S.M. Platelet endothelial cell adhesion molecule, PECAM-1, modulates cell migration. J Cell Physiol 153, 417-428 (1992).
15. Maisonpierre, P.C., et al. Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science 277, 55-60 (1997).
16. Coussens, L.M., et al. Inflammatory mast cells up-regulate angiogenesis during squamous epithelial carcinogenesis. Genes Dev 13, 1382-1397 (1999).
17. Ferrara, N. Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney Int 56, 794-814 (1999).
18. Morbidelli, L., et al. B1 receptor involvement in the effect of bradykinin on venular endothelial cell proliferation and potentiation of FGF-2 effects. Br J Pharmacol 124, 1286-1292 (1998).
19. Abdel-Malak, N.A., et al. Angiopoietin-1 promotes endothelial cell proliferation and migration through AP-1-dependent autocrine production of interleukin-8. Blood 111, 4145-4154 (2008).
20. Kishimoto, K., Liu, S., Tsuji, T., Olson, K.A. & Hu, G.F. Endogenous angiogenin in endothelial cells is a general requirement for cell proliferation and angiogenesis. Oncogene 24, 445-456 (2005).
21. Takehara, K., LeRoy, E.C. & Grotendorst, G.R. TGF-beta inhibition of endothelial cell proliferation: alteration of EGF binding and EGF-induced growth-regulatory (competence) gene expression. Cell 49, 415-422 (1987).
22. Gentilini, G., Kirschbaum, N.E., Augustine, J.A., Aster, R.H. & Visentin, G.P. Inhibition of human umbilical vein endothelial cell proliferation by the CXC chemokine, platelet factor 4 (PF4), is associated with impaired downregulation of p21(Cip1/WAF1). Blood 93, 25-33 (1999).
23. Tucci, M., et al. Modulation of insulin-like growth factor (IGF) and IGF binding protein biosynthesis by hypoxia in cultured vascular endothelial cells. J Endocrinol 157, 13-24 (1998).
24. Zhou, Z., Reddy, K., Guan, H. & Kleinerman, E.S. VEGF(165), but not VEGF(189), stimulates vasculogenesis and bone marrow cell migration into Ewing's sarcoma tumors in vivo. Mol Cancer Res 5, 1125-1132 (2007).
25. Nakatsu, M.N., et al. VEGF(121) and VEGF(165) regulate blood vessel diameter through vascular endothelial growth factor receptor 2 in an in vitro angiogenesis model. Lab Invest 83, 1873-1885 (2003).
26. Gamble, J.R., et al. Regulation of in vitro capillary tube formation by anti-integrin antibodies. J Cell Biol 121, 931-943 (1993).
27. Tolsma, S.S., Stack, M.S. & Bouck, N. Lumen formation and other angiogenic activities of cultured capillary endothelial cells are inhibited by thrombospondin-1. Microvasc Res 54, 13-26 (1997).
28. Kurz, H. Physiology of angiogenesis. J Neurooncol 50, 17-35 (2000).
29. Asahara, T., et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science 275, 964-967 (1997).
30. Shi, Q., et al. Evidence for circulating bone marrow-derived endothelial cells. Blood 92, 362-367 (1998).
31. Fuchs, E. & Segre, J.A. Stem cells: a new lease on life. Cell 100, 143-155 (2000).
32. Peichev, M., et al. Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors. Blood 95, 952-958 (2000).
33. Gehling, U.M., et al. In vitro differentiation of endothelial cells from AC133-positive progenitor cells. Blood 95, 3106-3112 (2000).
34. Urbich, C. & Dimmeler, S. Endothelial progenitor cells: characterization and role in vascular biology. Circ Res 95, 343-353 (2004).
35. Schmeisser, A., et al. Monocytes coexpress endothelial and macrophagocytic lineage markers and form cord-like structures in Matrigel under angiogenic conditions. Cardiovasc Res 49, 671-680 (2001).
36. Urbich, C., et al. Relevance of monocytic features for neovascularization capacity of circulating endothelial progenitor cells. Circulation 108, 2511-2516 (2003).
37. Hur, J., et al. Characterization of two types of endothelial progenitor cells and their different contributions to neovasculogenesis. Arterioscler Thromb Vasc Biol 24, 288-293 (2004).
38. Hirschi, K.K., Ingram, D.A. & Yoder, M.C. Assessing identity, phenotype, and fate of endothelial progenitor cells. Arterioscler Thromb Vasc Biol 28, 1584-1595 (2008).
39. Hill, J.M., et al. Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. N Engl J Med 348, 593-600 (2003).
40. Vasa, M., et al. Increase in circulating endothelial progenitor cells by statin therapy in patients with stable coronary artery disease. Circulation 103, 2885-2890 (2001).
41. De Palma, M., et al. Tie2 identifies a hematopoietic lineage of proangiogenic monocytes required for tumor vessel formation and a mesenchymal population of pericyte progenitors. Cancer Cell 8, 211-226 (2005).
42. Zentilin, L., et al. Bone marrow mononuclear cells are recruited to the sites of VEGF-induced neovascularization but are not incorporated into the newly formed vessels. Blood 107, 3546-3554 (2006).
43. Jin, D.K., et al. Cytokine-mediated deployment of SDF-1 induces revascularization through recruitment of CXCR4+ hemangiocytes. Nat Med 12, 557-567 (2006).
44. Yoder, M.C., et al. Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals. Blood 109, 1801-1809 (2007).
45. Isner, J.M. & Asahara, T. Angiogenesis and vasculogenesis as therapeutic strategies for postnatal neovascularization. J Clin Invest 103, 1231-1236 (1999).
46. Krause, D.S., et al. Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell. Cell 105, 369-377 (2001).
47. Mezey, E., Chandross, K.J., Harta, G., Maki, R.A. & McKercher, S.R. Turning blood into brain: cells bearing neuronal antigens generated in vivo from bone marrow. Science 290, 1779-1782 (2000).
48. Kawada, H. & Ogawa, M. Bone marrow origin of hematopoietic progenitors and stem cells in murine muscle. Blood 98, 2008-2013 (2001).
49. Yamamoto, K., et al. Molecular evaluation of endothelial progenitor cells in patients with ischemic limbs: therapeutic effect by stem cell transplantation. Arterioscler Thromb Vasc Biol 24, e192-196 (2004).
50. Badorff, C., et al. Transdifferentiation of blood-derived human adult endothelial progenitor cells into functionally active cardiomyocytes. Circulation 107, 1024-1032 (2003).
51. Zhang, Z.G., Zhang, L., Jiang, Q. & Chopp, M. Bone marrow-derived endothelial progenitor cells participate in cerebral neovascularization after focal cerebral ischemia in the adult mouse. Circ Res 90, 284-288 (2002).
52. Lee, I.G., Chae, S.L. & Kim, J.C. Involvement of circulating endothelial progenitor cells and vasculogenic factors in the pathogenesis of diabetic retinopathy. Eye (Lond) 20, 546-552 (2006).
53. Vajkoczy, P., et al. Multistep nature of microvascular recruitment of ex vivo-expanded embryonic endothelial progenitor cells during tumor angiogenesis. J Exp Med 197, 1755-1765 (2003).
54. Crosby, J.R., et al. Endothelial cells of hematopoietic origin make a significant contribution to adult blood vessel formation. Circ Res 87, 728-730 (2000).
55. Jackson, K.A., et al. Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells. J Clin Invest 107, 1395-1402 (2001).
56. Lyden, D., et al. Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med 7, 1194-1201 (2001).
57. Llevadot, J., et al. HMG-CoA reductase inhibitor mobilizes bone marrow--derived endothelial progenitor cells. J Clin Invest 108, 399-405 (2001).
58. Murayama, T., et al. Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo. Exp Hematol 30, 967-972 (2002).
59. Garcia-Barros, M., et al. Tumor response to radiotherapy regulated by endothelial cell apoptosis. Science 300, 1155-1159 (2003).
60. De Palma, M., Venneri, M.A., Roca, C. & Naldini, L. Targeting exogenous genes to tumor angiogenesis by transplantation of genetically modified hematopoietic stem cells. Nat Med 9, 789-795 (2003).
61. Theise, N.D., et al. Liver from bone marrow in humans. Hepatology 32, 11-16 (2000).
62. Theise, N.D., et al. Derivation of hepatocytes from bone marrow cells in mice after radiation-induced myeloablation. Hepatology 31, 235-240 (2000).
63. Gill, M., et al. Vascular trauma induces rapid but transient mobilization of VEGFR2(+)AC133(+) endothelial precursor cells. Circ Res 88, 167-174 (2001).
64. Orlic, D., et al. Bone marrow cells regenerate infarcted myocardium. Nature 410, 701-705 (2001).
65. Heissig, B., et al. Recruitment of stem and progenitor cells from the bone marrow niche requires MMP-9 mediated release of kit-ligand. Cell 109, 625-637 (2002).
66. Lee, S.H., et al. Early expression of angiogenesis factors in acute myocardial ischemia and infarction. N Engl J Med 342, 626-633 (2000).
67. Pillarisetti, K. & Gupta, S.K. Cloning and relative expression analysis of rat stromal cell derived factor-1 (SDF-1)1: SDF-1 alpha mRNA is selectively induced in rat model of myocardial infarction. Inflammation 25, 293-300 (2001).
68. Shintani, S., et al. Mobilization of endothelial progenitor cells in patients with acute myocardial infarction. Circulation 103, 2776-2779 (2001).
69. Takahashi, T., et al. Ischemia- and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med 5, 434-438 (1999).
70. Heeschen, C., et al. Erythropoietin is a potent physiologic stimulus for endothelial progenitor cell mobilization. Blood 102, 1340-1346 (2003).
71. Bahlmann, F.H., et al. Erythropoietin regulates endothelial progenitor cells. Blood 103, 921-926 (2004).
72. Aicher, A., et al. Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. Nat Med 9, 1370-1376 (2003).
73. Yamaguchi, J., et al. Stromal cell-derived factor-1 effects on ex vivo expanded endothelial progenitor cell recruitment for ischemic neovascularization. Circulation 107, 1322-1328 (2003).
74. Askari, A.T., et al. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy. Lancet 362, 697-703 (2003).
75. Kalka, C., et al. Vascular endothelial growth factor(165) gene transfer augments circulating endothelial progenitor cells in human subjects. Circ Res 86, 1198-1202 (2000).
76. Kuang, C.Y., et al. Knockdown of TRPC1 reduces the proliferation and migration of endothelial progenitor cells. Stem Cells Dev (2011).
77. Wang, H., et al. Inhibitor of DNA binding-1 promotes the migration and proliferation of endothelial progenitor cells in vitro. Mol Cell Biochem 335, 19-27 (2010).
78. Ferrara, N., et al. Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature 380, 439-442 (1996).
79. Fong, G.H., Rossant, J., Gertsenstein, M. & Breitman, M.L. Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 376, 66-70 (1995).
80. Dimmeler, S., et al. HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. J Clin Invest 108, 391-397 (2001).
81. Guo, Y., et al. The homeoprotein Hex is required for hemangioblast differentiation. Blood 102, 2428-2435 (2003).
82. Zippo, A., De Robertis, A., Bardelli, M., Galvagni, F. & Oliviero, S. Identification of Flk-1 target genes in vasculogenesis: Pim-1 is required for endothelial and mural cell differentiation in vitro. Blood 103, 4536-4544 (2004).
83. Frid, M.G., Kale, V.A. & Stenmark, K.R. Mature vascular endothelium can give rise to smooth muscle cells via endothelial-mesenchymal transdifferentiation: in vitro analysis. Circ Res 90, 1189-1196 (2002).
84. Hristov, M., Erl, W., Linder, S. & Weber, P.C. Apoptotic bodies from endothelial cells enhance the number and initiate the differentiation of human endothelial progenitor cells in vitro. Blood 104, 2761-2766 (2004).
85. Suh, W., et al. Transplantation of endothelial progenitor cells accelerates dermal wound healing with increased recruitment of monocytes/macrophages and neovascularization. Stem Cells 23, 1571-1578 (2005).
86. Li, B., et al. KDR (VEGF receptor 2) is the major mediator for the hypotensive effect of VEGF. Hypertension 39, 1095-1100 (2002).
87. Kawachi, Y., et al. Acute arterial thrombosis due to platelet aggregation in a patient receiving granulocyte colony-stimulating factor. Br J Haematol 94, 413-416 (1996).
88. Watt, S.M. & Fox, A. Blood vessel stem cells and wound healing. Br J Surg 92, 1461-1463 (2005).
89. Werner, N., et al. Circulating endothelial progenitor cells and cardiovascular outcomes. N Engl J Med 353, 999-1007 (2005).
90. Chen, P.L. & Easton, A.S. Evidence that tumor necrosis factor-related apoptosis inducing ligand (TRAIL) inhibits angiogenesis by inducing vascular endothelial cell apoptosis. Biochem Biophys Res Commun 391, 936-941 (2010).
91. Campochiaro, P.A. & Hackett, S.F. Ocular neovascularization: a valuable model system. Oncogene 22, 6537-6548 (2003).
92. Bikfalvi, A. [Tumor angiogenesis]. Bull Cancer 94, F193-198 (2007).
93. Guo, N., Krutzsch, H.C., Inman, J.K. & Roberts, D.D. Thrombospondin 1 and type I repeat peptides of thrombospondin 1 specifically induce apoptosis of endothelial cells. Cancer Res 57, 1735-1742 (1997).
94. Takahashi, S., Shinya, T. & Sugiyama, A. Angiostatin inhibition of vascular endothelial growth factor-stimulated nitric oxide production in endothelial cells. J Pharmacol Sci 112, 432-437 (2010).
95. Kim, I., Kim, J.H., Ryu, Y.S., Liu, M. & Koh, G.Y. Tumor necrosis factor-alpha upregulates angiopoietin-2 in human umbilical vein endothelial cells. Biochem Biophys Res Commun 269, 361-365 (2000).
96. Montesano, R. & Orci, L. Transforming growth factor beta stimulates collagen-matrix contraction by fibroblasts: implications for wound healing. Proc Natl Acad Sci U S A 85, 4894-4897 (1988).
97. Clark, R.A., Folkvord, J.M., Hart, C.E., Murray, M.J. & McPherson, J.M. Platelet isoforms of platelet-derived growth factor stimulate fibroblasts to contract collagen matrices. J Clin Invest 84, 1036-1040 (1989).
98. Schiro, J.A., et al. Integrin alpha 2 beta 1 (VLA-2) mediates reorganization and contraction of collagen matrices by human cells. Cell 67, 403-410 (1991).
99. Madlener, M., Parks, W.C. & Werner, S. Matrix metalloproteinases (MMPs) and their physiological inhibitors (TIMPs) are differentially expressed during excisional skin wound repair. Exp Cell Res 242, 201-210 (1998).
100. Levenson, S.M., et al. The Healing of Rat Skin Wounds. Ann Surg 161, 293-308 (1965).
101. Moritz, A.R. & Henriques, F.C. Studies of Thermal Injury: II. The Relative Importance of Time and Surface Temperature in the Causation of Cutaneous Burns. Am J Pathol 23, 695-720 (1947).
102. Arturson, M.G. The pathophysiology of severe thermal injury. J Burn Care Rehabil 6, 129-146 (1985).
103. Zawacki, B.E. Reversal of capillary stasis and prevention of necrosis in burns. Ann Surg 180, 98-102 (1974).
104. Jonsson, C.E., Granstrom, E. & Hamberg, M. Prostaglandins and thromboxanes in burn injury in man. Scand J Plast Reconstr Surg 13, 45-47 (1979).
105. Robson, M.C., DelBeccaro, E.J., Heggers, J.P. & Loy, G.L. Increasing dermal perfusion after burning by decreasing thromboxane production. J Trauma 20, 722-725 (1980).
106. Zawacki, B.E. The natural history of reversible burn injury. Surg Gynecol Obstet 139, 867-872 (1974).
107. Heimbach, D., Engrav, L., Grube, B. & Marvin, J. Burn depth: a review. World J Surg 16, 10-15 (1992).
108. Jackson, D.M. [The diagnosis of the depth of burning]. Br J Surg 40, 588-596 (1953).
109. Bajaj, S.P., Nield, D.V., Rayment, R. & Khoo, C.T. A simple modification of the pin-prick test for the assessment of burn depth in children. Burns Incl Therm Inj 14, 468-472 (1988).
110. Mileski, W.J., et al. Serial measurements increase the accuracy of laser Doppler assessment of burn wounds. J Burn Care Rehabil 24, 187-191 (2003).
111. Desai, M.H., Rutan, R.L. & Herndon, D.N. Conservative treatment of scald burns is superior to early excision. J Burn Care Rehabil 12, 482-484 (1991).
112. Still, J.M., Law, E.J., Klavuhn, K.G., Island, T.C. & Holtz, J.Z. Diagnosis of burn depth using laser-induced indocyanine green fluorescence: a preliminary clinical trial. Burns 27, 364-371 (2001).
113. Hlava, P., Moserova, J. & Konigova, R. Validity of clinical assessment of the depth of a thermal injury. Acta Chir Plast 25, 202-208 (1983).
114. Jeng, J.C., et al. Laser Doppler imaging determines need for excision and grafting in advance of clinical judgment: a prospective blinded trial. Burns 29, 665-670 (2003).
115. Jones, O.C., Wilson, D.I. & Andrews, S. The reliability of digital images when used to assess burn wounds. J Telemed Telecare 9 Suppl 1, S22-24 (2003).
116. Roa, L., Gomez-Cia, T., Acha, B. & Serrano, C. Digital imaging in remote diagnosis of burns. Burns 25, 617-623 (1999).
117. Droog, E.J., Steenbergen, W. & Sjoberg, F. Measurement of depth of burns by laser Doppler perfusion imaging. Burns 27, 561-568 (2001).
118. Watts, A.M., Tyler, M.P., Perry, M.E., Roberts, A.H. & McGrouther, D.A. Burn depth and its histological measurement. Burns 27, 154-160 (2001).
119. Chvapil, M., Speer, D.P., Owen, J.A. & Chvapil, T.A. Identification of the depth of burn injury by collagen stainability. Plast Reconstr Surg 73, 438-441 (1984).
120. Lawson, R.N. & Gaston, J.P. Temperature Measurements of Localized Pathological Processes. Ann N Y Acad Sci 121, 90-98 (1964).
121. Cole, R.P., Jones, S.G. & Shakespeare, P.G. Thermographic assessment of hand burns. Burns 16, 60-63 (1990).
122. Liddington, M.I. & Shakespeare, P.G. Timing of the thermographic assessment of burns. Burns 22, 26-28 (1996).
123. Zawacki, B.E. & Walker, H.L. An evaluation of patent blue V, bromphenol blue, and tetracycline for the diagnosis of burn depth. Plast Reconstr Surg 45, 459-465 (1970).
124. Dingwall, J.A. A Clinical Test for Differentiating Second from Third Degree Burns. Ann Surg 118, 427-429 (1943).
125. Gatti, J.E., LaRossa, D., Silverman, D.G. & Hartford, C.E. Evaluation of the burn wound with perfusion fluorometry. J Trauma 23, 202-206 (1983).
126. Black, K.S., et al. Burn depth evaluation with fluorometry: is it really definitive? J Burn Care Rehabil 7, 313-317 (1986).
127. Green, H.A., Bua, D., Anderson, R.R. & Nishioka, N.S. Burn depth estimation using indocyanine green fluorescence. Arch Dermatol 128, 43-49 (1992).
128. Holland, A.J., Martin, H.C. & Cass, D.T. Laser Doppler imaging prediction of burn wound outcome in children. Burns 28, 11-17 (2002).
129. Kamolz, L.P., et al. Indocyanine green video angiographies help to identify burns requiring operation. Burns 29, 785-791 (2003).
130. Sheridan, R.L., et al. Burn depth estimation by use of indocyanine green fluorescence: initial human trial. J Burn Care Rehabil 16, 602-604 (1995).
131. Haslik, W., et al. The influence of dressings and ointments on the qualitative and quantitative evaluation of burn wounds by ICG video-angiography: an experimental setup. Burns 30, 232-235 (2004).
132. Essex, T.J. & Byrne, P.O. A laser Doppler scanner for imaging blood flow in skin. J Biomed Eng 13, 189-194 (1991).
133. Micheels, J., Alsbjorn, B. & Sorensen, B. Laser doppler flowmetry. A new non-invasive measurement of microcirculation in intensive care? Resuscitation 12, 31-39 (1984).
134. Yeong, E.K., Mann, R., Goldberg, M., Engrav, L. & Heimbach, D. Improved accuracy of burn wound assessment using laser Doppler. J Trauma 40, 956-961; discussion 961-952 (1996).
135. Park, D.H., et al. Use of laser Doppler flowmetry for estimation of the depth of burns. Plast Reconstr Surg 101, 1516-1523 (1998).
136. O'Reilly, T.J., Spence, R.J., Taylor, R.M. & Scheulen, J.J. Laser Doppler flowmetry evaluation of burn wound depth. J Burn Care Rehabil 10, 1-6 (1989).
137. Bray, R., et al. Laser Doppler imaging of burn scars: a comparison of wavelength and scanning methods. Burns 29, 199-206 (2003).
138. Nanney, L.B., Wenczak, B.A. & Lynch, J.B. Progressive burn injury documented with vimentin immunostaining. J Burn Care Rehabil 17, 191-198 (1996).
139. Pape, S.A., Skouras, C.A. & Byrne, P.O. An audit of the use of laser Doppler imaging (LDI) in the assessment of burns of intermediate depth. Burns 27, 233-239 (2001).
140. Anselmo, V.J. & Zawacki, B.E. Multispectral photographic analysis. A new quantitative tool to assist in the early diagnosis of thermal burn depth. Ann Biomed Eng 5, 179-193 (1977).
141. Sayman, H.B., Demir, M., Cetinkale, O., Ayan, F. & Onsel, C. A method to evaluate microcirculatory vascular patency of full thickness burn in an animal model. Panminerva Med 41, 5-9 (1999).
142. Koruda, M.J., et al. Assessing burn wound depth using in vitro nuclear magnetic resonance (NMR). J Surg Res 40, 475-481 (1986).
143. Bauer, J.A. & Sauer, T. Cutaneous 10 MHz ultrasound B scan allows the quantitative assessment of burn depth. Burns Incl Therm Inj 15, 49-51 (1989).
144. Brink, J.A., et al. Quantitative assessment of burn injury in porcine skin with high-frequency ultrasonic imaging. Invest Radiol 21, 645-651 (1986).
145. Wachtel, T.L., Leopold, G.R., Frank, H.A. & Frank, D.H. B-mode ultrasonic echo determination of depth of thermal injury. Burns Incl Therm Inj 12, 432-437 (1986).
146. Iraniha, S., et al. Determination of burn depth with noncontact ultrasonography. J Burn Care Rehabil 21, 333-338 (2000).
147. Foster, F.S., Pavlin, C.J., Harasiewicz, K.A., Christopher, D.A. & Turnbull, D.H. Advances in ultrasound biomicroscopy. Ultrasound Med Biol 26, 1-27 (2000).
148. Goertz, D.E., et al. High-frequency color flow imaging of the microcirculation. Ultrasound Med Biol 26, 63-71 (2000).
149. Diaz, L.A. & Giudice, G.J. End of the century overview of skin blisters. Arch Dermatol 136, 106-112 (2000).
150. Lawrence, C. Treating minor burns. Nurs Times 85, 69-73 (1989).
151. Cope, O. The Treatment of the Surface Burns. Ann Surg 117, 885-893 (1943).
152. Gimbel, N.S., Kapetansky, D.I., Weissman, F. & Pinkus, H.K. A study of epithelization in blistered burns. AMA Arch Surg 74, 800-803 (1957).
153. Forage, A.V. The effects of removing the epidermis from burnt skin. Lancet 2, 690-693 (1962).
154. Singer, A.J., Thode, H.C., Jr. & McClain, S.A. The effects of epidermal debridement of partial-thickness burns on infection and reepithelialization in swine. Acad Emerg Med 7, 114-119 (2000).
155. Swain, A.H., Azadian, B.S., Wakeley, C.J. & Shakespeare, P.G. Management of blisters in minor burns. Br Med J (Clin Res Ed) 295, 181 (1987).
156. Geronemus, R.G. & Robins, P. The effect of two new dressings on epidermal wound healing. J Dermatol Surg Oncol 8, 850-852 (1982).
157. Uchinuma, E., Koganei, Y., Shioya, N. & Yoshizato, K. Biological evaluation of burn blister fluid. Ann Plast Surg 20, 225-230 (1988).
158. McKay, I.A. & Leigh, I.M. Epidermal cytokines and their roles in cutaneous wound healing. Br J Dermatol 124, 513-518 (1991).
159. Ono, I., Gunji, H., Suda, K., Iwatsuki, K. & Kaneko, F. Evaluation of cytokines in donor site wound fluids. Scand J Plast Reconstr Surg Hand Surg 28, 269-273 (1994).
160. Ono, I., Gunji, H., Zhang, J.Z., Maruyama, K. & Kaneko, F. A study of cytokines in burn blister fluid related to wound healing. Burns 21, 352-355 (1995).
161. Wilson, Y., et al. Investigation of the presence and role of calmodulin and other mitogens in human burn blister fluid. J Burn Care Rehabil 15, 303-314 (1994).
162. Honari, S. Topical therapies and antimicrobials in the management of burn wounds. Crit Care Nurs Clin North Am 16, 1-11 (2004).
163. Del Beccaro, E.J., Heggers, J.P. & Robson, M.C. Preventing the prostaglandin effect on dermal ischemia in the burn wound. Surg Forum 29, 603-605 (1978).
164. Rockwell, W.B. & Ehrlich, H.P. Fibrinolysis inhibition in human burn blister fluid. J Burn Care Rehabil 11, 1-6 (1990).
165. Haycock, J.W., Ralston, D.R., Morris, B., Freedlander, E. & MacNeil, S. Oxidative damage to protein and alterations to antioxidant levels in human cutaneous thermal injury. Burns 23, 533-540 (1997).
166. Deitch, E.A. & Smith, B.J. The effect of blister fluid from thermally injured patients on normal lymphocyte transformation. J Trauma 23, 106-110 (1983).
167. Kupper, T.S., Deitch, E.A., Baker, C.C. & Wong, W.C. The human burn wound as a primary source of interleukin-1 activity. Surgery 100, 409-415 (1986).
168. Deitch, E.A. & Emmett, M. Early protein alteration in blister fluid and serum associated with burn injury. J Trauma 26, 34-39 (1986).
169. Deitch, E.A., Bridges, R.M., Dobke, M. & McDonald, J.C. Burn wound sepsis may be promoted by a failure of local antibacterial host defenses. Ann Surg 206, 340-348 (1987).
170. Kagan, R.J. & Warden, G.D. Management of the burn wound. Clin Dermatol 12, 47-56 (1994).
171. Johnson, R.M. & Richard, R. Partial-thickness burns: identification and management. Adv Skin Wound Care 16, 178-187; quiz 188-179 (2003).
172. Clayton, M.C. & Solem, L.D. No ice, no butter. Advice on management of burns for primary care physicians. Postgrad Med 97, 151-155, 159-160, 165 (1995).
173. Duncan, D.J. & Driscoll, D.M. Burn wound management. Crit Care Nurs Clin North Am 3, 199-220 (1991).
174. Saranto, J.R., Rubayi, S. & Zawacki, B.E. Blisters, cooling, antithromboxanes, and healing in experimental zone-of-stasis burns. J Trauma 23, 927-933 (1983).
175. Garner, W.L., Zuccaro, C., Marcelo, C., Rodriguez, J.L. & Smith, D.J., Jr. The effects of burn blister fluid on keratinocyte replication and differentiation. J Burn Care Rehabil 14, 127-131 (1993).
176. Reagan, B.J., et al. The effects of burn blister fluid on cultured keratinocytes. J Trauma 40, 361-367 (1996).
177. Merz, J., et al. Wound care of the pediatric burn patient. AACN Clin Issues 14, 429-441 (2003).
178. Flanagan, M. & Graham, J. Should burn blisters be left intact or debrided? J Wound Care 10, 41-45 (2001).
179. Bishop, J.F. Burn wound assessment and surgical management. Crit Care Nurs Clin North Am 16, 145-177 (2004).
180. Collier, M. Wound bed preparation: theory to practice. Nurs Stand 17, 45-52; quiz 54-45 (2003).
181. Wilson, A.M., McGrouther, D.A., Eastwood, M. & Brown, R.A. The effect of burn blister fluid on fibroblast contraction. Burns 23, 306-312 (1997).
182. Liu, W., Wang, D.R. & Cao, Y.L. TGF-beta: a fibrotic factor in wound scarring and a potential target for anti-scarring gene therapy. Curr Gene Ther 4, 123-136 (2004).
183. Chung, W.H., et al. Granulysin is a key mediator for disseminated keratinocyte death in Stevens-Johnson syndrome and toxic epidermal necrolysis. Nat Med 14, 1343-1350 (2008).
184. Tepper, O.M., et al. Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation, adhesion, and incorporation into vascular structures. Circulation 106, 2781-2786 (2002).
185. Schmidt, A., Brixius, K. & Bloch, W. Endothelial precursor cell migration during vasculogenesis. Circ Res 101, 125-136 (2007).
186. Ferrara, N. & Davis-Smyth, T. The biology of vascular endothelial growth factor. Endocr Rev 18, 4-25 (1997).
187. Cribbs, R.K., Harding, P.A., Luquette, M.H. & Besner, G.E. Endogenous production of heparin-binding EGF-like growth factor during murine partial-thickness burn wound healing. J Burn Care Rehabil 23, 116-125 (2002).
188. Inoue, M., Zhou, L.J., Gunji, H., Ono, I. & Kaneko, F. Effects of cytokines in burn blister fluids on fibroblast proliferation and their inhibition with the use of neutralizing antibodies. Wound Repair Regen 4, 426-432 (1996).
189. Rehman, J., Li, J., Orschell, C.M. & March, K.L. Peripheral blood "endothelial progenitor cells" are derived from monocyte/macrophages and secrete angiogenic growth factors. Circulation 107, 1164-1169 (2003).
190. Zhang, S.J., et al. Adult endothelial progenitor cells from human peripheral blood maintain monocyte/macrophage function throughout in vitro culture. Cell Res 16, 577-584 (2006).
191. Atiyeh, B.S., Gunn, S.W. & Hayek, S.N. State of the art in burn treatment. World J Surg 29, 131-148 (2005).
192. Heil, M., et al. Blood monocyte concentration is critical for enhancement of collateral artery growth. Am J Physiol Heart Circ Physiol 283, H2411-2419 (2002).
193. Sandri, M., et al. Effects of exercise and ischemia on mobilization and functional activation of blood-derived progenitor cells in patients with ischemic syndromes: results of 3 randomized studies. Circulation 111, 3391-3399 (2005).
194. Dugan, A.L., et al. Serum levels of prolactin, growth hormone, and cortisol in burn patients: correlations with severity of burn, serum cytokine levels, and fatality. J Burn Care Rehabil 25, 306-313 (2004).
195. Grayson, L.S., et al. Quantitation of cytokine levels in skin graft donor site wound fluid. Burns 19, 401-405 (1993).
196. Holzheimer, R.G. & Steinmetz, W. Local and systemic concentrations of pro- and anti-inflammatory cytokines in human wounds. Eur J Med Res 5, 347-355 (2000).
197. Rockwell, W.B. & Ehrlich, H.P. Should burn blister fluid be evacuated? J Burn Care Rehabil 11, 93-95 (1990).
198. Sargent, R.L. Management of blisters in the partial-thickness burn: an integrative research review. J Burn Care Res 27, 66-81 (2006).
199. Rennekampff, H.O., et al. Bioactive interleukin-8 is expressed in wounds and enhances wound healing. J Surg Res 93, 41-54 (2000).
200. Tredget, E.E. The basis of fibrosis and wound healing disorders following thermal injury. J Trauma 62, S69 (2007).
201. Cohen, I.K. & McCoy, B.J. The biology and control of surface overhealing. World J Surg 4, 289-295 (1980).
202. Caulfield, R.H., Tyler, M.P., Austyn, J.M., Dziewulski, P. & McGrouther, D.A. The relationship between protease/anti-protease profile, angiogenesis and re-epithelialisation in acute burn wounds. Burns 34, 474-486 (2008).
203. Yang, L., et al. Identification of fibrocytes in postburn hypertrophic scar. Wound Repair Regen 13, 398-404 (2005).
204. Wu, W.S., Wang, F.S., Yang, K.D., Huang, C.C. & Kuo, Y.R. Dexamethasone induction of keloid regression through effective suppression of VEGF expression and keloid fibroblast proliferation. J Invest Dermatol 126, 1264-1271 (2006).
205. Monstrey, S., Hoeksema, H., Verbelen, J., Pirayesh, A. & Blondeel, P. Assessment of burn depth and burn wound healing potential. Burns 34, 761-769 (2008).
206. Riordan, C.L., et al. Noncontact laser Doppler imaging in burn depth analysis of the extremities. J Burn Care Rehabil 24, 177-186 (2003).
207. Avniel, S., et al. Involvement of the CXCL12/CXCR4 pathway in the recovery of skin following burns. J Invest Dermatol 126, 468-476 (2006).
208. McCarthy, D.W., et al. Production of heparin-binding epidermal growth factor-like growth factor (HB-EGF) at sites of thermal injury in pediatric patients. J Invest Dermatol 106, 49-56 (1996).
209. Nissen, N.N., Gamelli, R.L., Polverini, P.J. & DiPietro, L.A. Differential angiogenic and proliferative activity of surgical and burn wound fluids. J Trauma 54, 1205-1210; discussion 1211 (2003).
210. Pan, S.C., Wu, L.W., Chen, C.L., Shieh, S.J. & Chiu, H.Y. Deep partial thickness burn blister fluid promotes neovascularization in the early stage of burn wound healing. Wound Repair Regen 18, 311-318 (2010).
211. Rafii, S. & Lyden, D. Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat Med 9, 702-712 (2003).
212. Ceradini, D.J. & Gurtner, G.C. Homing to hypoxia: HIF-1 as a mediator of progenitor cell recruitment to injured tissue. Trends Cardiovasc Med 15, 57-63 (2005).
213. Wu, Y., Zhao, R.C. & Tredget, E.E. Concise review: bone marrow-derived stem/progenitor cells in cutaneous repair and regeneration. Stem Cells 28, 905-915 (2010).
214. Fett, J.W., et al. Isolation and characterization of angiogenin, an angiogenic protein from human carcinoma cells. Biochemistry 24, 5480-5486 (1985).
215. Strydom, D.J. The angiogenins. Cell Mol Life Sci 54, 811-824 (1998).
216. Liote, F., Champy, R., Moenner, M., Boval-Boizard, B. & Badet, J. Elevated angiogenin levels in synovial fluid from patients with inflammatory arthritis and secretion of angiogenin by cultured synovial fibroblasts. Clin Exp Immunol 132, 163-168 (2003).
217. Nanney, L.B. Epidermal and dermal effects of epidermal growth factor during wound repair. J Invest Dermatol 94, 624-629 (1990).
218. Arenberg DA, Keane MP, DiGiovine B, Kunkel SL, Morris SB, Xue YY, Burdick MD, Glass MC, Iannettoni MD, Strieter RM. Epithelial-neutrophil activating peptide (ENA-78) is an important angiogenic factor in non-small cell lung cancer. J Clin Invest 1998;102:465-72.
219. Koch AE, Volin MV, Woods JM, Kunkel SL, Connors MA, Harlow LA, Woodruff DC, Burdick MD, Strieter RM. Regulation of angiogenesis by the C-X-C chemokines interleukin-8 and epithelial neutrophil activating peptide 78 in the rheumatoid joint. Arthritis Rheum 2001;44:31-40.
220. Adams, S.A. & Subramanian, V. The angiogenins: an emerging family of ribonuclease related proteins with diverse cellular functions. Angiogenesis 3, 189-199 (1999).
221. Shimoyama, S., Yamasaki, K., Kawahara, M. & Kaminishi, M. Increased serum angiogenin concentration in colorectal cancer is correlated with cancer progression. Clin Cancer Res 5, 1125-1130 (1999).
222. Hartmann, A., et al. Hypoxia-induced up-regulation of angiogenin in human malignant melanoma. Cancer Res 59, 1578-1583 (1999).
223. Olson, K.A., Verselis, S.J. & Fett, J.W. Angiogenin is regulated in vivo as an acute phase protein. Biochem Biophys Res Commun 242, 480-483 (1998).
224. Galiano, R.D., et al. Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells. Am J Pathol 164, 1935-1947 (2004).
225. Chan, D.A., et al. Tumor vasculature is regulated by PHD2-mediated angiogenesis and bone marrow-derived cell recruitment. Cancer Cell 15, 527-538 (2009).
226. Hoeksema, H., et al. Accuracy of early burn depth assessment by laser Doppler imaging on different days post burn. Burns 35, 36-45 (2009).
227. Zhang, X., et al. Association of increasing burn severity in mice with delayed mobilization of circulating angiogenic cells. Arch Surg 145, 259-266 (2010).
228. Gangemi, E.N., Carnino, R. & Stella, M. Videocapillaroscopy in postburn scars: in vivo analysis of the microcirculation. Burns 36, 799-805 (2010).
229. Kischer, C.W. The microvessels in hypertrophic scars, keloids and related lesions: a review. J Submicrosc Cytol Pathol 24, 281-296 (1992).
230. Yang, L., et al. Peripheral blood fibrocytes from burn patients: identification and quantification of fibrocytes in adherent cells cultured from peripheral blood mononuclear cells. Lab Invest 82, 1183-1192 (2002).
231. Wang, J., Chen, H., Shankowsky, H.A., Scott, P.G. & Tredget, E.E. Improved scar in postburn patients following interferon-alpha2b treatment is associated with decreased angiogenesis mediated by vascular endothelial cell growth factor. J Interferon Cytokine Res 28, 423-434 (2008).