Transplantation of BMSCs expressing hPDGF-A/hBD2 promotes wound healing in rats with combined radiation-wound injury
Open Access
- 14 August 2008
- journal article
- research article
- Published by Springer Science and Business Media LLC in Gene Therapy
- Vol. 16 (1), 34-42
- https://doi.org/10.1038/gt.2008.133
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 (Pin 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 (Pgroup 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.Keywords
This publication has 28 references indexed in Scilit:
- Bone marrow‐derived mesenchymal stromal cells accelerate wound healing in the ratWound Repair and Regeneration, 2006
- First experience in the use of bone marrow mesenchymal stem cells for the treatment of a patient with deep skin burnsBulletin of Experimental Biology and Medicine, 2005
- Mesenchymal stem cells and haematopoietic stem cell transplantationBest Practice & Research Clinical Haematology, 2004
- Mesenchymal stem cells and haematopoietic stem cell transplantationBest Practice & Research Clinical Haematology, 2004
- Adenovirus Encoding Human Platelet-Derived Growth Factor-B Delivered in Collagen Exhibits Safety, Biodistribution, and Immunogenicity Profiles Favorable for Clinical UseMolecular Therapy, 2004
- HLA expression and immunologic propertiesof differentiated and undifferentiated mesenchymal stem cellsExperimental Hematology, 2003
- Human dermal microvascular endothelial cells form vascular analogs in cultured skin substitutes after grafting to athymic miceThe FASEB Journal, 2002
- Mesenchymal stem cellsJournal of Orthopaedic Research, 1991
- Growth factor-induced acceleration of tissue repair through direct and inductive activities in a rabbit dermal ulcer model.JCI Insight, 1991
- Differential Staining of Collagens Type I, II and III by Sirius Red and Polarization MicroscopyArchivum histologicum japonicum, 1978