3D scaffold development for tissue engineering

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Πάντσιος, Πασχάλης
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During this thesis a novel perfusion bioreactor was constructed. Based on an existing bioreactor apparatus [1], investigating previous researches and using spare parts found in the Laboratory the bioreactor was designed and assembled. It was tested for functionality and non-toxicity by culturing cells in it and it passed. In association with Biomedical Research Foundations (Academy of Athens), Human Umbilical Artery specimens were tested. The ulterior purpose is to use HUA as grafts. Mesenchymal Stem Cells cultivated in decellularized HUA in incubator for one day and in the perfusion bioreactor for five days. The main parameter controlled was the flow rate, depending on the shear stress of the fluid (culture medium) flow. The results were significant. A successful recellularization was accomplished with a high cell density on the lumen of arteries. The results are depicted implementing Hematoxylin & Eosin Stain and confocal microscopy techniques processed at the BRFAA. The second experimental series consisted of ten-layered Polycaprolactone-Carbon Nanotubes scaffolds, manufactured in our Laboratory using a prototype Electrospinning unit. PCL-CNT scaffolds are considered promising tool for osteogenesis. Perfusion bioreactor MSC cultures, halting at one and three days, were compared to static cultures applying MTT assay. A strong indication was deducted that perfusion is quite more efficient to the proliferation of MSC, than in the case of static culture. Unfortunately, the time margins were narrow enough to be an obstacle to a more thorough investigation of the researches above. More days of culture and repetitive experiments are considered mandatory for a complete investigation. Finally, the next step should be the investigation of MSC differentiation to either endothelial cells, or osteocytes, regarding HUA and PCL-CNT scaffolds, respectively.
Bioreactors, Tissue engineering