Cell and Tissue Research

, Volume 329, Issue 2, pp 301–311 | Cite as

Treatment of diabetic wounds with fetal murine mesenchymal stromal cells enhances wound closure

  • Andrea T. Badillo
  • Robert A. Redden
  • Liping Zhang
  • Edward J. Doolin
  • Kenneth W. Liechty
Regular Article

Abstract

Diabetes impairs multiple aspects of the wound-healing response. Delayed wound healing continues to be a significant healthcare problem for which effective therapies are lacking. We have hypothesized that local delivery of mesenchymal stromal cells (MSC) at a wound might correct many of the wound-healing impairments seen in diabetic lesions. We treated excisional wounds of genetically diabetic (Db-/Db-) mice and heterozygous controls with either MSC, CD45+ cells, or vehicle. At 7 days, treatment with MSC resulted in a decrease in the epithelial gap from 3.2 ± 0.5 mm in vehicle-treated wounds to 1.3 ± 0.4 mm in MSC-treated wounds and an increase in granulation tissue from 0.8 ± 0.3 mm2 to 2.4 ± 0.6 mm2, respectively (mean ± SD, P < 0.04). MSC-treated wounds also displayed a higher density of CD31+ vessels and exhibited increases in the production of mRNA for epidermal growth factor, transforming growth factor beta 1, vascular endothelial growth factor, and stromal-derived growth factor 1-alpha. MSC also demonstrated greater contractile ability than fibroblast controls in a collagen gel contraction assay. The effects of locally applied MSC are thus sufficient to improve healing in diabetic mice. Possible mechanisms of this effect include augmented local growth-factor production, improved neovascularization, enhanced cellular recruitment to wounds, and improved wound contraction.

Keywords

Stromal progenitor cells Mesenchymal stromal cells Mesenchymal stem cells Wound healing Diabetes Mouse (C57BKS strains) 

References

  1. Akino K, Mineda T, Akita S (2005) Early cellular changes of human mesenchymal stem cells and their interaction with other cells. Wound Repair Regen 13:434–440PubMedCrossRefGoogle Scholar
  2. Anker PS in ’t, Noort WA, Scherjon SA, Kleijburg-van der Keur C, Kruisselbrink AB, Bezooijen RL van, Beekhuizen W, Willemze R, Kanhai HH, Fibbe WE (2003) Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential. Haematologica 88:845–852Google Scholar
  3. Badiavas EV, Falanga V (2003) Treatment of chronic wounds with bone marrow-derived cells. Arch Dermatol 139:510–516PubMedCrossRefGoogle Scholar
  4. Badiavas EV, Abedi M, Butmarc J, Falanga V, Quesenberry P (2003) Participation of bone marrow derived cells in cutaneous wound healing. J Cell Physiol 196:245–250PubMedCrossRefGoogle Scholar
  5. Bhakta S, Hong P, Koc O (2006) The surface adhesion molecule cxcr4 stimulates mesenchymal stem cell migration to stromal cell-derived factor–1 in vitro but does not decrease apoptosis under serum deprivation. Cardiovasc Revasc Med 7:19–24PubMedCrossRefGoogle Scholar
  6. Blakytny R, Jude E (2006) The molecular biology of chronic wounds and delayed healing in diabetes. Diabet Med 23:594–608PubMedCrossRefGoogle Scholar
  7. Broughton G 2nd, Janis JE, Attinger CE (2006) The basic science of wound healing. Plast Reconstr Surg 117:12S–34SPubMedCrossRefGoogle Scholar
  8. Ceradini DJ, Kulkarni AR, Callaghan MJ, Tepper OM, Bastidas N, Kleinman ME, Capla JM, Galiano RD, Levine JP, Gurtner GC (2004) Progenitor cell trafficking is regulated by hypoxic gradients through hif–1 induction of SDF–1. Nat Med 10:858–864PubMedCrossRefGoogle Scholar
  9. Dar A, Kollet O, Lapidot T (2006) Mutual, reciprocal sdf–1/cxcr4 interactions between hematopoietic and bone marrow stromal cells regulate human stem cell migration and development in nod/scid chimeric mice. Exp Hematol 34:967–975PubMedCrossRefGoogle Scholar
  10. Digirolamo CM, Stokes D, Colter D, Phinney DG, Class R, Prockop DJ (1999) Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol 107:275–281PubMedCrossRefGoogle Scholar
  11. Dinh TL, Veves A (2006) The efficacy of apligraf in the treatment of diabetic foot ulcers. Plast Reconstr Surg 117:152S–157SPubMedCrossRefGoogle Scholar
  12. Falanga V (2005) Wound healing and its impairment in the diabetic foot. Lancet 366:1736–1743PubMedCrossRefGoogle Scholar
  13. Fathke C, Wilson L, Hutter J, Kapoor V, Smith A, Hocking A, Isik F (2004) Contribution of bone marrow-derived cells to skin: collagen deposition and wound repair. Stem Cells 22:812–822PubMedCrossRefGoogle Scholar
  14. Fedyk ER, Jones D, Critchley HO, Phipps RP, Blieden TM, Springer TA (2001) Expression of stromal-derived factor–1 is decreased by IL–1 and TNF and in dermal wound healing. J Immunol 166:5749–5754PubMedGoogle Scholar
  15. Gang EJ, Jeong JA, Han S, Yan Q, Jeon CJ, Kim H (2006) In vitro endothelial potential of human uc blood-derived mesenchymal stem cells. Cytotherapy 8:215–227PubMedCrossRefGoogle Scholar
  16. Javazon EH, Colter DC, Schwarz EJ, Prockop DJ (2001) Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells. Stem Cells 19:219–225PubMedCrossRefGoogle Scholar
  17. Kern S, Eichler H, Stoeve J, Kluter H, Bieback K (2006) Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells 24:1294–1301PubMedCrossRefGoogle Scholar
  18. Kim DH, Yoo KH, Choi KS, Choi J, Choi SY, Yang SE, Yang YS, Im HJ, Kim KH, Jung HL, Sung KW, Koo HH (2005) Gene expression profile of cytokine and growth factor during differentiation of bone marrow-derived mesenchymal stem cell. Cytokine 31:119–126PubMedCrossRefGoogle Scholar
  19. Kinnaird T, Stabile E, Burnett MS, Shou M, Lee CW, Barr S, Fuchs S, Epstein SE (2004) Local delivery of marrow-derived stromal cells augments collateral perfusion through paracrine mechanisms. Circulation 109:1543–1549PubMedCrossRefGoogle Scholar
  20. Liechty KW, MacKenzie TC, Shaaban AF, Radu A, Moseley AM, Deans R, Marshak DR, Flake AW (2000) Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep. Nat Med 6:1282–1286PubMedCrossRefGoogle Scholar
  21. Nagaya N, Fujii T, Iwase T, Ohgushi H, Itoh T, Uematsu M, Yamagishi M, Mori H, Kangawa K, Kitamura S (2004) Intravenous administration of mesenchymal stem cells improves cardiac function in rats with acute myocardial infarction through angiogenesis and myogenesis. Am J Physiol Heart Circ Physiol 287:H2670–H2676PubMedCrossRefGoogle Scholar
  22. Opalenik SR, Davidson JM (2005) Fibroblast differentiation of bone marrow-derived cells during wound repair. FASEB J 19:1561–1563PubMedGoogle Scholar
  23. Oswald J, Boxberger S, Jorgensen B, Feldmann S, Ehninger G, Bornhauser M, Werner C (2004) Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells 22:377–384PubMedCrossRefGoogle Scholar
  24. Pandit A, Ashar R, Feldman D (1999) The effect of TGF-beta delivered through a collagen scaffold on wound healing. J Invest Surg 12:89–100PubMedCrossRefGoogle Scholar
  25. Parikka V, Vaananen A, Risteli J, Salo T, Sorsa T, Vaananen HK, Lehenkari P (2005) Human mesenchymal stem cell derived osteoblasts degrade organic bone matrix in vitro by matrix metalloproteinases. Matrix Biol 24:438–447PubMedCrossRefGoogle Scholar
  26. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147PubMedCrossRefGoogle Scholar
  27. Redden RA, Doolin EJ (2006) Complementary roles of microtubules and microfilaments in the lung fibroblast-mediated contraction of collagen gels: dynamics and the influence of cell density. In Vitro Cell Dev Biol Anim 42:70–74PubMedCrossRefGoogle Scholar
  28. Silva Meirelles L da, Chagastelles PC, Nardi NB (2006) Mesenchymal stem cells reside in virtually all post-natal organs and tissues. J Cell Sci 119:2204–2213PubMedCrossRefGoogle Scholar
  29. Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341:738–746PubMedCrossRefGoogle Scholar
  30. Sullivan SR, Underwood RA, Gibran NS, Sigle RO, Usui ML, Carter WG, Olerud JE (2004) Validation of a model for the study of multiple wounds in the diabetic mouse (Db/Db). Plast Reconstr Surg 113:953–960PubMedCrossRefGoogle Scholar
  31. Sung HJ, Meredith C, Johnson C, Galis ZS (2004) The effect of scaffold degradation rate on three-dimensional cell growth and angiogenesis. Biomaterials 25:5735–5742PubMedCrossRefGoogle Scholar
  32. Tropel P, Noel D, Platet N, Legrand P, Benabid AL, Berger F (2004) Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow. Exp Cell Res 295:395–406PubMedCrossRefGoogle Scholar
  33. Veves A, Falanga V, Armstrong DG, Sabolinski ML (2001) Graftskin, a human skin equivalent, is effective in the management of noninfected neuropathic diabetic foot ulcers: a prospective randomized multicenter clinical trial. Diabetes Care 24:290–295PubMedCrossRefGoogle Scholar
  34. Wang M, Crisostomo PR, Herring C, Meldrum KK, Meldrum DR (2006) Human progenitor cells from bone marrow or adipose tissue produce VEGF, HGF, and IGF-I in response to TNF by a p38 MAPK-dependent mechanism. Am J Physiol Regul Integr Comp Physiol 291:R880–R884PubMedGoogle Scholar
  35. Wieman TJ, Smiell JM, Su Y (1998) Efficacy and safety of a topical gel formulation of recombinant human platelet-derived growth factor-BB (becaplermin) in patients with chronic neuropathic diabetic ulcers. A phase III randomized placebo-controlled double-blind study. Diabetes Care 21:822–827PubMedCrossRefGoogle Scholar
  36. Wu GD, Nolta JA, Jin YS, Barr ML, Yu H, Starnes VA, Cramer DV (2003) Migration of mesenchymal stem cells to heart allografts during chronic rejection. Transplantation 75:679–685PubMedCrossRefGoogle Scholar
  37. Wu S, Suzuki Y, Ejiri Y, Noda T, Bai H, Kitada M, Kataoka K, Ohta M, Chou H, Ide C (2003) Bone marrow stromal cells enhance differentiation of cocultured neurosphere cells and promote regeneration of injured spinal cord. J Neurosci Res 72:343–351PubMedCrossRefGoogle Scholar
  38. Wu M, Yang L, Liu S, Li H, Hui N, Wang F, Liu H (2006) Differentiation potential of human embryonic mesenchymal stem cells for skin-related tissue. Br J Dermatol 155:282–291PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Andrea T. Badillo
    • 1
  • Robert A. Redden
    • 1
  • Liping Zhang
    • 1
  • Edward J. Doolin
    • 1
  • Kenneth W. Liechty
    • 1
    • 2
  1. 1.The Center for Fetal Research at The Children’s Hospital of PhiladelphiaThe University of Pennsylvania School of MedicinePhiladelphiaUSA
  2. 2.Abramson Research CenterPhiladelphiaUSA

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