Artikel
In vivo use of bone marrow derived cells to improve vascularization in a dermal matrix regeneration model
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Autoren
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J.T. Egana
- Dept. Plastic and Hand Surgery, Burn Center, UKSH Campus Lübeck, Lübeck, Germany
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F.A. Fierro
- Technical University of Dresden, Dresden, Germany
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M. Butzal
- UKSH Campus Lübeck, Lübeck, Germany
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N. Bauer
- Technical University of Dresden, Dresden, Germany
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S. Lavandero
- FONDAP Center for Molecular Studies of the Cell, University of Chile. Santiago, Chile
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S. Krüger
- UKSH Campus Lübeck, Lübeck, Germany
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P. Schlenke
- UKSH Campus Lübeck, Lübeck, Germany
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H.G. Machens
- Dept. Plastic and Hand Surgery, Burn Center, UKSH Campus Lübeck, Lübeck, Germany
| Veröffentlicht: | 1. Oktober 2007 |
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Gliederung
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Introduction: Neovascularization plays a critical role during matrix based dermal regeneration, since blood vessels have to supply nutrients, oxygen and immunologically competent cells for the tissue in regeneration to ensure therapeutic success. Bone marrow derived stem cells have not yet been employed for dermal matrix bioactivation. In this work we combined the use of a bioartificial template for dermal regeneration with bone marrow derived cells to improve neodermal vascularization in vivo.
Materials and methods: Apheresis samples were obtained from G-CSF mobilized human donors. Total bone marrow derived cells (TBMDC) were isolated by ficoll gradients and early hematopoietic stem cells (CD133+) were isolated by the use of magnetic microbeads. Cells were characterized by FACS. After that, each 250,000 cells were seeded in a matrix (Integra)for dermal regeneration of bilateral 15 mm diameter full skin defects of nu/nu mice (TBMDC in Group A, CD 133+ in Group B). Viability was analysed by MTT assay. Control matrices received cell medium only (Group C). Matrices were analysed after 2 weeks for total amount of angiogenesis. Each subgroup comprised 6 animals.
Results: CD133+ cells were isolated and FACS results showed more than 95% of puricity in the sample. In vitro, distribution of the cells was analyzed by confocal microscopy and interaction with the matrix was shown by SEM. MTT assay showed that both cell populations can survive, for at least two weeks in normal culture conditions. After 2 weeks of transplantation in a full skin defect model, microangiographical matrix analysis showed that both TPBMNC and CD133+ cells significantly improved vascularization levels compared to control matrices (P < 0.05). No statistical differences were found between both Groups A and B.
Discussion: In this work we show for the first time that direct engraftment of total human derived bone marrow mononuclear cells or early hematopoietic stem cells significantly improve neodermal vascularization in vivo. These results suggest that stem cell therapy could be used to develop new therapeutical approaches to improve vascularization during matrix based dermal regeneration.
