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Genetically-engineered bone marrow stromal cells and collagen mimetic scaffold modification for healing critically-sized bone defectsWojtowicz, Abigail M. 07 July 2009 (has links)
Non-healing bone defects have a significant socioeconomic impact in the U.S. with approximately 600,000 bone grafting procedures performed annually. Autografts and allografts are clinically the most common treatments; however, autologous donor bone is in limited supply, and allografts often have poor mechanical properties. Therefore, tissue engineering and regenerative medicine strategies are being developed to address issues with clinical bone grafting. The overall objective of this work was to develop bone tissue engineering strategies that enhance healing of orthotopic defects by targeting specific osteogenic cell signaling pathways. The general approach included the investigation of two different tissue engineering strategies, which both focused on directed osteoblastic differentiation to promote bone formation.
In the first cell-based strategy, we hypothesized that constitutive overexpression of the osteoblast-specific transcription factor, Runx2, in bone marrow stromal cells (BMSCs) would promote orthotopic bone formation in vivo. We tested this hypothesis by delivering Runx2-modified BMSCs on synthetic scaffolds to critically-sized defects in rats. We found that Runx2-modified BMSCs significantly increased orthotopic bone formation compared to empty defects, cell-free scaffolds and unmodified BMSCs. This gene therapy approach to bone regeneration provides a mineralizing cell source which has clinical relevance.
In the second biomaterial-based strategy, we hypothesized that incorporation of the collagen-mimetic peptide, GFOGER, into synthetic bone scaffolds would promote orthotopic bone formation in vivo without the use of cells or growth factors. We tested this hypothesis by passively adsorbing GFOGER onto poly-caprolactone (PCL) scaffolds and implanting them into critically-sized orthotopic defects in rats. We found that GFOGER-coated scaffolds significantly increased bone formation compared to uncoated scaffolds in a dose dependent manner. Development of this cell-free strategy for bone tissue engineering provides an inexpensive therapeutic alternative to clinical bone defect healing, which could be implemented as a point of care application.
Both strategies developed in this work take advantage of specific osteoblastic signaling pathways involved in bone healing. Further development of these tissue engineering strategies for bone regeneration will provide clinically-relevant treatment options for healing large bone defects in humans by employing well-controlled signals to promote bone formation and eliminating the need for donor bone.
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Utilisation de cellules souches médullaires en bioingénierie tissulaire du ligament / Use of bone marrow mesenchymal stem cell in bioengineering of ligamentsZhang, Lei 24 January 2008 (has links)
Les ligaments jouent un rôle important dans le mouvement et la stabilité des articulations. Les accidents et la fatigue chronique sont les principales raisons des ruptures de ligaments qui n’ont généralement pas de capacité de guérison, ce qui conduit à de graves dysfonctionnements du ligament et des articulations. La construction des ligaments en bioingénierie donne un nouvel espoir thérapeutique. Pour construire un tel tissu, les cellules sont très importantes dans la mise en oeuvre de la construction d’un biotissu ayant de bonnes propriétés tant biologiques que mécaniques. Quelle source cellulaire et quel microenvironnement doivent être utilisés pour la reconstruction des ligaments ? L’objectif de ce travail était d’étudier la différenciation des cellules souches mésenchymateuses médullaires (CSMM) en fibroblaste. Pour ce faire, d’une part, nous avons co-cultivé des CSMM du rat sans contact avec des fibroblastes ligamentaires, et d’autre part, nous avons stimulé mécaniquement les CSMM. Un suivi des ARNm et des protéines associées caractéristiques des ligaments (collagènes I et III et ténascine-C) a été analysés. Nos résultats expérimentaux ont montré que la culture des CSMM dans un microenvironnement fibroblastique de ligaments ou l’étirement favorisent les synthèses de collagènes I et III et de ténascine-C dans les proportions proches des ligaments. L’ensemble de cette étude suggère qu’il est envisageable d’utiliser les CSMM comme source cellulaire, pour une application clinique, en ingénierie tissulaire du ligaments / Ligaments play an important role in the movement and stability of joints. Accidents and chronic fatigues are the main reasons for ligament lesion which usually is difficult for self healing and leads to serious dysfunction of ligaments and joints. The construction of bioengineering ligaments gives a new way to overcome this problem. Cells are very important in the construction of a biotissue with appropriate biological as well as mechanical properties. Which cellular source and microenvironment should be used for the reconstruction of ligaments? The objective of this work is to study the differentiation of bone marrow mesenchymal stem cell (BMSC) into fibroblast. We co-cultured indirectly rat BMSC with ligament fibroblasts or stimulated them by mechanical stretching. After that, the expressions of characteristics mRNA and protein of ligaments (collagen I, III and tenascin-C) have been analyzed. Our experimental results showed that the culture of BMSC in a microenvironment of ligament fibroblast or under stretching favored the syntheses of collagen I, III and tenascin-C in the proportions close to ligaments. In summary, these studies suggest it is feasible to use BMSC as cellular source for a clinical application in tissue engineering of ligaments
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CD31(-) HipOps - A Highly Osteogenic Cell Population From Mouse Bone MarrowMcKenzie, Kristen Penny 04 December 2012 (has links)
Multipotent mesenchymal stem cells (MSCs), found in many adult tissues, may be useful for regenerative medicine applications. Their identification and purification have been difficult due to their low frequency and lack of unambiguous markers. Using a magnetic micro-beads negative selection technique to remove contaminating hematopoietic cells from mouse bone marrow stromal cells (BMSCs), our lab recently isolated a highly purified osteoprogenitor (HipOp) population that was also enriched for other mesenchymal precursors, including MSCs (Itoh and Aubin, 2009). To further enhance enrichment, we positively selected BMSCs and HipOps for CD73, a putative MSC marker, which resulted in no significant additional enrichment for osteoprogenitors when the population was tested in vitro. However, we also found that HipOps were enriched in vascular endothelial cells, and that removing these cells by further negative selection with CD31/PECAM resulted in a CD31(-) HipOp population with higher osteogenic capacity than HipOps in vitro and in vivo.
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CD31(-) HipOps - A Highly Osteogenic Cell Population From Mouse Bone MarrowMcKenzie, Kristen Penny 04 December 2012 (has links)
Multipotent mesenchymal stem cells (MSCs), found in many adult tissues, may be useful for regenerative medicine applications. Their identification and purification have been difficult due to their low frequency and lack of unambiguous markers. Using a magnetic micro-beads negative selection technique to remove contaminating hematopoietic cells from mouse bone marrow stromal cells (BMSCs), our lab recently isolated a highly purified osteoprogenitor (HipOp) population that was also enriched for other mesenchymal precursors, including MSCs (Itoh and Aubin, 2009). To further enhance enrichment, we positively selected BMSCs and HipOps for CD73, a putative MSC marker, which resulted in no significant additional enrichment for osteoprogenitors when the population was tested in vitro. However, we also found that HipOps were enriched in vascular endothelial cells, and that removing these cells by further negative selection with CD31/PECAM resulted in a CD31(-) HipOp population with higher osteogenic capacity than HipOps in vitro and in vivo.
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Dancing in borrowed shoes : a history of ballroom dancing in South Africa (1600s-1940s)Green, Alida Maria 20 October 2009 (has links)
This study deals with the history of ballroom dancing in South Africa. While reference will be made to the founding of ballroom in the early eighteenth century in South Africa, the study will mainly focus on the period between 1920 and 1940 in the Johannesburg, Pretoria region. The study will determine how and why ballroom dancing came to South Africa from abroad; how South Africans borrowed from the international dancing world; what they copied, what ideas they followed, how they chose to dance ballroom and how this affected South African society at large. Copyright / Dissertation (MHCS)--University of Pretoria, 2009. / Historical and Heritage Studies / unrestricted
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