Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Transplantation of BMSCs expressing hPDGF-A/hBD2 promotes wound healing in rats with combined radiation-wound injury

Gene Therapy volume 16, pages 34–42 (2009)Cite this article

Abstract

The aim of this study was to test the efficacy of transplantation of bone marrow-derived mesenchymal stem cells (BMSCs) expressing human platelet-derived growth factor A (hPDGF-A) and human beta-defensin2 (hBD2) in accelerating wound healing of combined radiation-wound injury. Recombinant adenovirus vector simultaneously expressing hPDGF-A and hBD2 was constructed and packaged into virus particles that were used to infect rat BMSCs. The expressions of the exogenous in BMSCs were determined by reverse transcriptase (RT)-PCR and western –blot, whereas the functions were determined by cell counting kit (CCK), wound-healing assay on monolayer cells and Kleihauer-Betke (K-B) test. The recombinant adenovirus-infected BMSCs (1 × 107) were subcutaneously transplanted into the wound bed and wound healing was observed for the indicated duration. Rats with combined total body ionizing radiation (6 Gy) and full-thickness skin excision (2% of total body surface area) wound injury were treated with normal BMSCs (group N), BMSCs infected with recombinant adenovirus expressing hPDGF-A and hBD2 (group T) or phosphate-buffered saline (PBS) (group S). The mean wound healing time, percentage of residual wound area (n=8), blind pathological observation (n=3 per time point for each group) and the amount of bacteria under the scar (the same sample was used in the pathological study, n=3) were used for evaluating wound healing. Collagen was visualized by Sirius red staining. Exogenous hPDGF-A and hBD2 were expressed in BMSCs as indicated by RT-PCR and western blot. Faster wound healing of scratched monolayer cells was demonstrated in hPDGF-A/hBD2 gene-modified BMSCs (T-MSCs) when compared with the corresponding control (P<0.01), and conditioned culture medium from T-MSCs showed stimulative effect on BMSC proliferation and in vitro antibiotic effect in the presence of trypsin. Neutralizing antibody interfering in vitro demonstrated that secreted hPDGF-A was the main factor stimulating cell proliferation. In an in vivo test, the radiation-wound combined injury exhibited shorter healing time (21 days). Histologically, there was better granulation formation/maturation and skin-dependent regeneration, as well as more collagen deposition (P<0.01) in rats of group T than in other groups. The deposition and remodeling of collagen in wounds were ranked in the following order: group T>group N>group S. Significantly less bacterial colony formation in the cultured under-scar samples in the rats of group T was observed (P<0.01) at day 7 and thereafter when compared with control. After transplantation, the BMSCs expressed exogenous genes in the wound for at least 2 weeks, as indicated by the reporter gene. Topical transplantation of gene-modified BMSCs promoted wound healing, which may be the benefit of the secretion of antibacterial hBD2 and mitogenic hPDGF-A, resulting in better granulation formation/maturation and skin appendage regeneration in wound. These data demonstrated the potential application of this combination of cell therapy and gene therapy on refractory wound healing.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Pierce GF, Tarpley JE, Allman RM, Goode PS, Serdar CM, Morris B et al. Tissue repair processes in healing chronic pressure ulcers treated with recombinant platelet-derived growth factor BB. Am J Pathol 1994; 145: 1399–1410.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Loots MA, Lamme EN, Zeegelaar J, Mekkes JR, Bos JD, Middelkoop E . Differences in cellular infiltrate and extracellular matrix of chronic diabetic and venous ulcers versus acute wounds. J Invest Dermatol 1998; 111: 850–857.

    Article  CAS  Google Scholar 

  3. Supp DM, Wilson-Landy K, Boyce ST . Human dermal microvascular endothelial cells form vascular analogs in cultured skin substitutes after grafting to athymic mice. FASEB J 2002; 16: 797–804.

    Article  CAS  Google Scholar 

  4. Robson MC . Wound infection. A failure of wound healing caused by an imbalance of bacteria. Surg Clin North Am 1997; 77: 637–650.

    Article  CAS  Google Scholar 

  5. Robson MC, Heggers JP . Bacterial quantification of open wounds. Mil Med 1969; 134: 19–24.

    Article  CAS  Google Scholar 

  6. Robson MC, Stenberg BD, Heggers JP . Wound healing alterations caused by infection. Clin Plast Surg 1990; 17: 485–492.

    CAS  PubMed  Google Scholar 

  7. Cooper R, Lawrence JC . Micro-organisms and wounds. J Wound Care 1996; 5: 233–236.

    Article  CAS  Google Scholar 

  8. Rasulov MF, Vasilchenkov AV, Onishchenko NA, Krasheninnikov ME, Kravchenko VI, Gorshenin TL et al. First experience of the use bone marrow mesenchymal stem cells for the treatment of a patient with deep skin burns. Bull Exp Biol Med 2005; 139: 141–144.

    Article  CAS  Google Scholar 

  9. McFarlin K, Gao X, Liu YB, Dulchavsky DS, Kwon D, Arbab AS et al. Bone marrow-derived mesenchymal stromal cells accelerate wound healing in the rat. Wound Repair Regen 2006; 14: 471–478.

    Article  Google Scholar 

  10. Caplan AI . Mesenchymal stem cells. J Orthop Res 1991; 9: 641–650.

    Article  CAS  Google Scholar 

  11. Prockop DJ . Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 1997; 276: 71–74.

    Article  CAS  Google Scholar 

  12. Haynesworth SE, Baber MA, Caplan AI . Cytokine expression by human marrow-derived mesenchymal progenitor cells in vitro: effects of dexamethasone and IL-1 alpha. J Cell Physiol 1996; 166: 585–592.

    Article  CAS  Google Scholar 

  13. Majumdar MK, Thiede MA, Mosca JD, Moorman M, Gerson SL . Phenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells. J Cell Physiol 1998; 176: 57–66.

    Article  CAS  Google Scholar 

  14. Le Blanc K, Tammik C, Rosendahl K, Zetterberg E, Ringden O . HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. Exp Hematol 2003; 31: 890–896.

    Article  CAS  Google Scholar 

  15. Bacigalupo A . Mesenchymal stem cells and haematopoietic stem cell transplantation. Best Pract Res Clin Haematol 2004; 17: 387–399.

    Article  CAS  Google Scholar 

  16. Mustoe TA, Pierce GF, Morishima C, Deuel TF . Growth factor-induced acceleration of tissue repair through direct and inductive activities in a rabbit dermal ulcer model. J Clin Invest 1991; 87: 694–703.

    Article  CAS  Google Scholar 

  17. Grotendorst GR, Martin GR, Pencev D, Sodek J, Harvey AK . Stimulation of granulation tissue formation by platelet-derived growth factor in normal and diabetic rats. J Clin Invest 1985; 76: 2323–2329.

    Article  CAS  Google Scholar 

  18. Albertson S, Hummel III RP, Breeden M, Greenhalgh DG . PDGF and FGF reverse the healing impairment in protein-malnourished diabetic mice. Surgery 1993; 114: 368–372.

    CAS  PubMed  Google Scholar 

  19. Mustoe TA, Purdy J, Gramates P, Deuel TF, Thomason A, Pierce GF . Reversal of impaired wound healing in irradiated rats by platelet-derived growth factor-BB. Am J Surg 1989; 158: 345–350.

    Article  CAS  Google Scholar 

  20. Harder J, Bartels J, Christophers E, Schroder JM . A peptide antibiotic from human skin. Nature 1997; 387: 861.

    Article  CAS  Google Scholar 

  21. Chadebech P, Goidin D, Jacquet C, Viac J, Schmitt D, Staquet MJ . Use of human reconstructed epidermis to analyze the regulation of beta-defensin hBD-1, hBD-2, and hBD-3 expression in response to LPS. Cell Biol Toxicol 2003; 19: 313–324.

    Article  CAS  Google Scholar 

  22. Chunmeng S, Tianmin C, Yongping S, Xinze R, Yue M, Jifu Q et al. Effects of dermal multipotent cell transplantation on skin wound healing. J Surg Res 2004; 121: 13–19.

    Article  Google Scholar 

  23. Gu DL, Nguyen T, Gonzalez AM, Printz MA, Pierce GF, Sosnowski BA et al. Adenovirus encoding human platelet-derived growth factor-B delivered in collagen exhibits safety, biodistribution, and immunogenicity profiles favorable for clinical use. Mol Ther 2004; 9: 699–711.

    Article  CAS  Google Scholar 

  24. Badiavas EV, Falanga V . Gene therapy. J Dermatol 2001; 28: 175–192.

    Article  CAS  Google Scholar 

  25. Crombleholme TM . Adenoviral-mediated gene transfer in wound healing. Wound Repair Regen 2000; 8: 460–472.

    Article  CAS  Google Scholar 

  26. Javazon EH, Colter DC, Schwarz EJ, Prockop DJ . 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 2001; 19: 219–225.

    Article  CAS  Google Scholar 

  27. Junqueira LC, Cossermelli W, Brentani R . Differential staining of collagens type I, II and III by Sirius red and polarization microscopy. Arch Histol Jpn 1978; 41: 267–274.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by the National Key Basic Research and Development Plan, China (‘973’ project 2005GB522605) and the Key Scientific Research Project of PLA, China (06G076 and 06H029).

Author information

Authors and Affiliations

  1. Department of Radiation Medicine, Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Third Military Medical University, Chongqing, China

    L Hao, J Wang, Z Zou, G Yan, S Dong, J Deng, X Ran, Y Feng, C Luo & T Cheng

  2. Department of Radiation Oncology, Kimmel Cancer Center of Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA, USA

    Y Wang

Authors
  1. L Hao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Z Zou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. X Ran
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Y Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. C Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Y Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. T Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Z Zou.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hao, L., Wang, J., Zou, Z. et al. Transplantation of BMSCs expressing hPDGF-A/hBD2 promotes wound healing in rats with combined radiation-wound injury. Gene Ther 16, 34–42 (2009). https://doi.org/10.1038/gt.2008.133

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gt.2008.133

Keywords

This article is cited by

Search

Advanced search

Quick links