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Angiogenesis

, Volume 16, Issue 4, pp 745–757 | Cite as

Enhanced wound vascularization using a dsASCs seeded FPEG scaffold

  • David O. Zamora
  • Shanmugasundaram Natesan
  • Sandra Becerra
  • Nicole Wrice
  • Eunna Chung
  • Laura J. Suggs
  • Robert J. ChristyEmail author
Original Paper

Abstract

The bioengineering of autologous vascular networks is of great importance in wound healing. Adipose-derived stem cells (ASCs) are of interest due to their ability to differentiate toward various cell types, including vascular. We hypothesized that adult human ASCs embedded in a three-dimensional PEG-fibrin (FPEG) gel have the ability to modulate vascularization of a healing wound. Initial in vitro characterization of ASCs isolated from discarded burn skin samples (dsASCs) and embedded in FPEG gels indicated they could express such pericyte/smooth muscle cell markers as α-smooth muscle actin, platelet-derived growth factor receptor-β, NG2 proteoglycan, and angiopoietin-1, suggesting that these cells could potentially be involved in a supportive cell role (i.e., pericyte/mural cell) for blood vessels. Using a rat skin excision model, wounds treated with dsASCs-FPEG gels showed earlier collagen deposition and wound remodeling compared to vehicle FPEG treated wounds. Furthermore, the dsASCs-seeded gels increased the number of vessels in the wound per square millimeter by day 16 (~66.7 vs. ~36.9/mm2) in these same studies. dsASCs may support this increase in vascularization through their trophic contribution of vascular endothelial growth factor, as determined by in vitro analysis of mRNA and the protein levels. Immunohistochemistry showed that dsASCs were localized to the surrounding regions of large blood-perfused vessels. Human dsASCs may play a supportive role in the formation of vascular structures in the healing wound through direct mechanisms as well as indirect trophic effects. The merging of autologous grafts or bioengineered composites with the host’s vasculature is critical, and the use of autologous dsASCs in these procedures may prove to be therapeutic.

Keywords

Angiogenesis ASCs Wound healing Fibrin Collagen PEG 

Notes

Acknowledgments

Dr. Natesan is supported by a Postdoctoral Fellowship Grant from the Pittsburgh Tissue Engineering Initiative (PTEI). Funding for this work was provided by the TATRC Foundation and the Deployment Related Medical Research Program.

Conflict of interest

The opinions or assertions contained herein are the private views of RJC and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense.

References

  1. 1.
    Chan RK et al (2012) Development of a vascularized skin construct using adipose-derived stem cells from debrided burned skin. Stem Cells Int 2012:841203PubMedGoogle Scholar
  2. 2.
    Owens BD et al (2008) Combat wounds in operation Iraqi Freedom and operation Enduring Freedom. J Trauma 64(2):295–299PubMedCrossRefGoogle Scholar
  3. 3.
    Wolf SE et al (2006) Comparison between civilian burns and combat burns from Operation Iraqi Freedom and Operation Enduring Freedom. Ann Surg 243(6):786–792; discussion 792-5PubMedCrossRefGoogle Scholar
  4. 4.
    Brusselaers N et al (2010) Skin replacement in burn wounds. J Trauma 68(2):490–501PubMedCrossRefGoogle Scholar
  5. 5.
    Carmeliet P, Jain RK (2000) Angiogenesis in cancer and other diseases. Nature 407(6801):249–257PubMedCrossRefGoogle Scholar
  6. 6.
    Rouwkema J, Rivron NC, van Blitterswijk CA (2008) Vascularization in tissue engineering. Trends Biotechnol 26(8):434–441PubMedCrossRefGoogle Scholar
  7. 7.
    Sahota PS et al (2003) Development of a reconstructed human skin model for angiogenesis. Wound Repair Regen 11(4):275–284PubMedCrossRefGoogle Scholar
  8. 8.
    Sorrell JM, Baber MA, Caplan AI (2007) A self-assembled fibroblast-endothelial cell co-culture system that supports in vitro vasculogenesis by both human umbilical vein endothelial cells and human dermal microvascular endothelial cells. Cells Tissues Organs 186(3):157–168PubMedCrossRefGoogle Scholar
  9. 9.
    Schechner JS et al (2003) Engraftment of a vascularized human skin equivalent. FASEB J 17(15):2250–2256PubMedCrossRefGoogle Scholar
  10. 10.
    Shepherd BR et al (2006) Vascularization and engraftment of a human skin substitute using circulating progenitor cell-derived endothelial cells. FASEB J 20(10):1739–1741PubMedCrossRefGoogle Scholar
  11. 11.
    Zhang CP, Fu XB (2008) Therapeutic potential of stem cells in skin repair and regeneration. Chin J Traumatol 11(4):209–221PubMedGoogle Scholar
  12. 12.
    Girandon L et al (2011) In vitro models for adipose tissue engineering with adipose-derived stem cells using different scaffolds of natural origin. Folia Biol (Praha) 57(2):47–56Google Scholar
  13. 13.
    Zhang G et al (2006) A PEGylated fibrin patch for mesenchymal stem cell delivery. Tissue Eng 12(1):9–19PubMedCrossRefGoogle Scholar
  14. 14.
    Liu H, Collins SF, Suggs LJ (2006) Three-dimensional culture for expansion and differentiation of mouse embryonic stem cells. Biomaterials 27(36):6004–6014PubMedCrossRefGoogle Scholar
  15. 15.
    Ahmann KA et al (2010) Fibrin degradation enhances vascular smooth muscle cell proliferation and matrix deposition in fibrin-based tissue constructs fabricated in vitro. Tissue Eng Part A 16(10):3261–3270PubMedCrossRefGoogle Scholar
  16. 16.
    Chalupowicz DG et al (1995) Fibrin II induces endothelial cell capillary tube formation. J Cell Biol 130(1):207–215PubMedCrossRefGoogle Scholar
  17. 17.
    Zuk PA et al (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13(12):4279–4295PubMedCrossRefGoogle Scholar
  18. 18.
    Hong SJ, Traktuev DO, March KL (2010) Therapeutic potential of adipose-derived stem cells in vascular growth and tissue repair. Curr Opin Organ Transplant 15(1):86–91PubMedCrossRefGoogle Scholar
  19. 19.
    Kim Y et al (2007) Direct comparison of human mesenchymal stem cells derived from adipose tissues and bone marrow in mediating neovascularization in response to vascular ischemia. Cell Physiol Biochem 20(6):867–876PubMedCrossRefGoogle Scholar
  20. 20.
    Moon MH et al (2006) Human adipose tissue-derived mesenchymal stem cells improve postnatal neovascularization in a mouse model of hindlimb ischemia. Cell Physiol Biochem 17(5–6):279–290PubMedCrossRefGoogle Scholar
  21. 21.
    Kondo K et al (2009) Implantation of adipose-derived regenerative cells enhances ischemia-induced angiogenesis. Arterioscler Thromb Vasc Biol 29(1):61–66PubMedCrossRefGoogle Scholar
  22. 22.
    Meruane MA, Rojas M, Marcelain K (2012) The use of adipose tissue-derived stem cells within a dermal substitute improves skin regeneration by increasing neoangiogenesis and collagen synthesis. Plast Reconstr Surg 130(1):53–63PubMedCrossRefGoogle Scholar
  23. 23.
    Blanton MW et al (2009) Adipose stromal cells and platelet-rich plasma therapies synergistically increase revascularization during wound healing. Plast Reconstr Surg 123(2 Suppl):56S–64SPubMedGoogle Scholar
  24. 24.
    Lu F et al (2008) Improved viability of random pattern skin flaps through the use of adipose-derived stem cells. Plast Reconstr Surg 121(1):50–58PubMedCrossRefGoogle Scholar
  25. 25.
    Natesan S et al (2011) Debrided skin as a source of autologous stem cells for wound repair. Stem Cells 29(8):1219–1230PubMedCrossRefGoogle Scholar
  26. 26.
    Natesan S et al (2011) A bilayer construct controls adipose-derived stem cell differentiation into endothelial cells and pericytes without growth factor stimulation. Tissue Eng Part A 17(7–8):941–953PubMedCrossRefGoogle Scholar
  27. 27.
    Cao Y et al (2005) Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Biochem Biophys Res Commun 332(2):370–379PubMedCrossRefGoogle Scholar
  28. 28.
    Fischer LJ et al (2009) Endothelial differentiation of adipose-derived stem cells: effects of endothelial cell growth supplement and shear force. J Surg Res 152(1):157–166PubMedCrossRefGoogle Scholar
  29. 29.
    da Silva Meirelles L, Caplan AI, Nardi NB (2008) In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26(9):2287–2299PubMedCrossRefGoogle Scholar
  30. 30.
    Gimble JM, Katz AJ, Bunnell BA (2007) Adipose-derived stem cells for regenerative medicine. Circ Res 100(9):1249–1260PubMedCrossRefGoogle Scholar
  31. 31.
    Rehman J et al (2004) Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation 109(10):1292–1298PubMedCrossRefGoogle Scholar
  32. 32.
    Rubina K et al (2009) Adipose stromal cells stimulate angiogenesis via promoting progenitor cell differentiation, secretion of angiogenic factors, and enhancing vessel maturation. Tissue Eng Part A 15(8):2039–2050PubMedCrossRefGoogle Scholar
  33. 33.
    Song SY, Chung HM, Sung JH (2010) The pivotal role of VEGF in adipose-derived-stem-cell-mediated regeneration. Expert Opin Biol Ther 10(11):1529–1537PubMedCrossRefGoogle Scholar
  34. 34.
    Brem H et al (2009) Mechanism of sustained release of vascular endothelial growth factor in accelerating experimental diabetic healing. J Invest Dermatol 129(9):2275–2287PubMedCrossRefGoogle Scholar
  35. 35.
    Wise LM et al (2012) The vascular endothelial growth factor (VEGF)-E encoded by orf virus regulates keratinocyte proliferation and migration and promotes epidermal regeneration. Cell Microbiol 14(9):1376–1390PubMedCrossRefGoogle Scholar
  36. 36.
    Kidoya H et al (2008) Spatial and temporal role of the apelin/APJ system in the caliber size regulation of blood vessels during angiogenesis. EMBO J 27(3):522–534PubMedCrossRefGoogle Scholar
  37. 37.
    Chiu LL, Radisic M (2010) Scaffolds with covalently immobilized VEGF and Angiopoietin-1 for vascularization of engineered tissues. Biomaterials 31(2):226–241PubMedCrossRefGoogle Scholar
  38. 38.
    Janmey PA, Winer JP, Weisel JW (2009) Fibrin gels and their clinical and bioengineering applications. J R Soc Interface 6(30):1–10PubMedCrossRefGoogle Scholar
  39. 39.
    van Hinsbergh VW, Collen A, Koolwijk P (2001) Role of fibrin matrix in angiogenesis. Ann N Y Acad Sci 936:426–437PubMedCrossRefGoogle Scholar
  40. 40.
    Lesman A et al (2011) Engineering vessel-like networks within multicellular fibrin-based constructs. Biomaterials 32(31):7856–7869PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht (outside the USA)  2013

Authors and Affiliations

  • David O. Zamora
    • 1
  • Shanmugasundaram Natesan
    • 1
  • Sandra Becerra
    • 1
  • Nicole Wrice
    • 1
  • Eunna Chung
    • 2
  • Laura J. Suggs
    • 2
  • Robert J. Christy
    • 1
    Email author
  1. 1.Regenerative Medicine Research ProgramUnited States Army Institute of Surgical ResearchFort Sam HoustonUSA
  2. 2.Department of Biomedical EngineeringThe University of Texas at AustinAustinUSA

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