Columbus, OH; 2Veterinary Biosciences, The Ohio State. University, Columbus, OH. Approximately six million fractures occur yearly in the United. States. Ten to ...
STEM CELL GENE THERAPY were upregulated in MSCs, compared to cardiac tissue, Feridex by LCM and MSCs in culture (cathepsins, cystatins and matrix metalloprotease 9 in particular). Feridex labeling of MSCs may affect cell growth, differentiation and apoptosis. Gene expression profiles of injected MSCs obtained by LCM may be less useful to assess structural cardiac genes, due to surrounding tissue background, but may reveal biologic pathways influenced by cell injection.
1043. Intravenous (I.V.) Administration of Manganese Superoxide Dismutase-Plasmid Liposome (MnSOD-PL) Complex Protects Mice from Whole Body Irradiation Michael W. Epperly,1 Yunyun Niu,1 Joel S. Greenberger.1 1 Radiation Oncology, University of Pittsburgh Cancer Institute, Pittsburgh, PA. Local administration of MnSOD-PL has been demonstrated to protect normal tissues from irradiation damage including: oral cavity, esophagus, lung, bladder and intestines. To determine whether systemic administration of MnSOD-PL protected the bone marrow from irradiation-induced damage, C57BL/6NHsd female mice were injected intravenously with MnSOD-PL (100 µg plasmid DNA). Groups of the mice were irradiated 24 hours later to 9 Gy, 9.5 Gy or 9.75 Gy whole body irradiation. Following irradiation, the mice were weighed on a daily basis and watched for signs of irradiationinduced hematologic damage. Control mice irradiated to either 9.5 Gy or 9.75 Gy demonstrated a loss of body weight while mice injected with the MnSOD-PL showed no changes in body weight. Mice injected with MnSOD-PL before irradiation demonstrated protection against irradiation with stabilized body weight and increased survival compared to control irradiated mice (p=0.0402 at 9.5 Gy or p = 0.0016 at 9.75 Gy). Therefore, I.V. administration of MnSOD-PL protects mice from bone marrow toxicity of whole body irradiation. Joel S. Greenberger, MD is a co-owner of and holds equity in Automated Cell, Inc., Pittsburgh, PA.
(NoAd) or transduced with AdBMP2 or Ad-luciferase reporter gene construct (AdLuc), suspended in 50 µl of 1.2% ALG. Controls were empty ALG and untreated osteotomies. Differentiation of BMDMSC in vitro was monitored in ALG constructs using realtime RT-PCR to document changes in gene expression of aggrecan, type II collagen, type I collagen, and BMP2. Luciferase expression was monitored using an in vivo imaging system. Healing was compared using quantitative micro-computed tomography (microCT), fluorescent labeling, and histology at post-operative day 14. Upregulation of aggrecan, type II collagen, type I collagen, and BMP2 gene expression were confirmed in BMDMSC transduced with AdBMP2 and suspended in ALG (AdBMP2xBMDMSC/ ALG). Osteotomy gap area (mm2) was greater (p < 0.01) in the AdBMP2xBMDMSC/ALG group (Figure 1) when compared with untreated osteotomies (Figure 2). In all ALG groups, but not in untreated osteotomies, bone healing was impeded by development of a cartilage mass. Untreated osteotomy gaps filled with fibrous tissue and bone. While this was an unexpected finding, a threedimensional alginate matrix prevented healing of bone and cartilage. Further study is warranted, particularly in immunocompetent animal models, to evaluate other delivery methods for genetically modified BMDMSC as treatments for complex articular fractures.
1044. Mesenchymal Stem Cell-Mediated Gene Delivery of Bone Morphogenetic Protein-2 in Alginate in an Articular Fracture Model Terri A. Zachos,1 Steven E. Weisbrode,2 Alicia L. Bertone.1,2 1 Veterinary Clinical Sciences, The Ohio State University, Columbus, OH; 2Veterinary Biosciences, The Ohio State University, Columbus, OH. Approximately six million fractures occur yearly in the United States. Ten to 15% of these will become delayed or nonunions, resulting in prolonged pain and frequently surgery, increasing the monetary burden on the health care system. Articular fractures are particularly problematic. Gene therapy may represent a viable treatment option. The purpose of our study was to create a clinically relevant animal model of articular fracture healing, and to evaluate use of genetically modified bone marrow-derived mesenchymal stem cells (BMDMSC) to heal bone and cartilage. Our hypothesis was that BMDMSC genetically modified to express bone morphogenetic protein (BMP)-2, delivered in a three-dimensional alginate (ALG) matrix, would augment healing, when compared with wild-type cells in ALG. A first-generation adenoviral vector (Ad) was used to deliver human BMP2 cDNA (AdBMP2) to BMDMSC from Lewis rats (from Tulane Center for Gene Therapy). Articular osteotomies were created in nude rats and treated with BMDMSC, either wild-type
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Molecular Therapy Volume 13, Supplement 1, May 2006 Copyright The American Society of Gene Therapy
