Osteogenic differentiation as a result of BMP-2 plasmid DNA based gene therapy in vitro and in vivo
| Autor: | Fiona Wegman, F. C. Oner, Leontine Schuijff, Wouter J.A. Dhert, A Bijenhof, Jacqueline Alblas |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Stromal cell
lcsh:Diseases of the musculoskeletal system Alginates Cellular differentiation Green Fluorescent Proteins bone graft lcsh:Surgery Bone Morphogenetic Protein 2 Mice Nude osteogenic differentiation Bone morphogenetic protein (BMP) Bone morphogenetic protein 2 Prosthesis Implantation Mice Calcification Physiologic Glucuronic Acid bone regeneration In vivo Osteogenesis Cell Line Tumor Paracrine Communication multipotent stromal cells (MSC) Animals Humans Bone regeneration biology Goats Hexuronic Acids Cell Differentiation Mesenchymal Stem Cells Transfection DNA Genetic Therapy lcsh:RD1-811 Alkaline Phosphatase Flow Cytometry Molecular biology transfection Naked DNA Osteocalcin biology.protein hydrogel lcsh:RC925-935 Plasmids |
| Zdroj: | European Cells & Materials, Vol 21, Pp 230-241 (2011) Europe PubMed Central |
| ISSN: | 1473-2262 |
| Popis: | Bone regeneration is one of the major focus points in the field of regenerative medicine. A well-known stimulus of bone formation is bone morphogenetic protein-2 (BMP-2), which has already been extensively used in clinical applications. We investigated the possibility of achieving osteogenic differentiation both in vitro and in vivo as a result of prolonged presence of BMP-2 using plasmid DNA-based gene therapy. By delivering BMP-2 cDNA in an alginate hydrogel, a versatile formulation is developed. High transfection efficiencies of up to 95% were obtained in both human multipotent stromal cells (MSCs) and MG-63 cells using naked DNA in vitro. Over a period of 5 weeks, an increasing amount of biologically active BMP-2 was released from the cells and remained present in the gel. In vivo, transfected cells were found after both two and six weeks implantation in naked mice, even in groups without seeded cells, thus indicating in vivo transfection of endogenous cells. The protein levels were effective in inducing osteogenic differentiation in vitro, as seen by elevated alkaline phosphatase (ALP) production and in vivo, as demonstrated by the production of collagen I and osteocalcin in a mineralised alginate matrix. We conclude that BMP-2 cDNA incorporated in alginate hydrogel appears to be a promising new strategy for minimal-invasive delivery of growth factors in bone regeneration. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|