O-43: Effects of TGF-b and b-FGF on Potential of Peripheral Blood-Borne Stem Cells and Bone Marrow-Derived Stem Cells in Wound Healing in a Murine Model
Objective: Peripheral blood fibrocytes are a newly identified leukocyte subpopulation that displays fibroblast-like properties. These blood-borne cells can rapidly enter the site of injury at the same time as circulating inflammatory cells. Marrow stroma includes a subpopulation of undifferentiated cells that are capable of becoming one of a number of phenotypes, including chondrocytes, osteoblasts, adipocytes, fibroblasts. Adult human bone marrow contains a minority population of MSCs that contribute to the regeneration of tissues such as bone, cartilage, muscle, ligaments, tendons, fat, and stroma. Evidence that these MSCs are pluripotent, rather than being a mixture of committed progenitor cells each with a restricted potential, includes their rapid proliferation in culture. We hypotheses that peripheral blood Mesenchymal Stem cells and bone marrow Mesenchymal Stem Cells have effective role in wound healing. In this study we identified and quantified the MSCs derived from Blood and Bone Marrow recruited and migrated to wound site. Materials and Methods: - Cell isolation from peripheral blood and culturing - Cell isolation from bone marrow and culturing - Proliferation assay - Migration assay - RT-PCR for type I collagen mRNA Results: Our results showed Synergistic effect of TGF-B and b-B-FGF leads to a significant increase in migration and recruitment of both PBMSC and BMSCs to the wound site with more potent effect on PBMSc compared to BMSCs. RT-PCR amplification of collagen type I transcripts (348 bp) confirmed that TGF-b and b-FGF activate the Col-1 production in MSCs at transcription level with more vigorous effect of TGF-b on PB-MSCs compared to same condition on BMSCs. Conclusion: TGF-b has role in fibroblast proliferation and collagen production . Basic fibroblast growth factor (b-FGF) plays a pivotal role in wound repair. b-FGF necessity for normal wound repair has been confirmed by several workers. We found that b-FGF alone is not able to increase the number of PBMCs and BMSCs recruited to the wound site, but significantly improved cell migration to the wound site when treated with TGF-b suggesting prerequisite role of TGF-b in stem cell differentiation and localization. Our findings are the first to indicate that TGF-b and b-FGF has synergistic effect in mesenchymal stem cell recruitment in vivo and that this is more potent in peripheral blood mesenchymal stem cell recruitment than bone marrow mesenchymal stem cells.