Doctor of Philosophy (PhD)



Document Type



Adult multipotent stromal cells (MSCs) in combination with biocompatible scaffolds may augment well-established medical and surgical techniques. Pre-implantation cell loading and viability on scaffold carriers followed by in vitro differentiation and extracellular matrix (ECM) formation contribute to the efficacy of in vivo tissue formation. Given the specificity of the parameters for each cell-scaffold construct, it is vital to assess the relative potential of various combinations to select the best option for various clinical needs. High demand for equine and human bone regeneration especially non-union fracture repair is still largely unaddressed. Therefore osteogenesis potential of both equine and human MSCs engrafted on bioscaffolds was comprehensively investigated in this study. After loading onto scaffolds, cells attached to scaffolds were assessed immediately and following different time of culture. Cell number, viability, distribution and differentiation were evaluated with DNA levels, confocal laser, light, and scanning electron microscopy (SEM) and RT-PCR. Both perfusion bioreactor and spinner flasks loading proved to be reliable and efficient procedures for seeding cells within bioscaffolds as well as achieving uniform cell distribution. Tissue specific micro- and ultrastructural changes were evident in equine cell-scaffold constructs cultured in induction medium. The versatility and suitability of type I collagen (COLI) scaffolds for equine adult MSC tissue regeneration was confirmed by generation of distinct mesenchymal tissues by paired adipose- (ASCs) and bone marrow derived multipotent stromal cells (BMSCs). In vivo study of equine MSCs on COLI scaffold further supported bone formation by BMSC constructs after short-term induction. However, ASC constructs produced comparative ECM as that of BMSC constructs but under commitment into dermal-like tissue. Pre-induction times as well as osteoinductive biomaterial addition to ColI scaffold were optimized to improve efficiency of human ASCs constructs to commit osteogenesis. The customized bioscaffold was proved in vitro as a promising carrier of MSCs to meet diverse clinical needs for osteogenesis. Overall, this research established straightforward and reproducible procedures to support customized tissue engineering with bioreactor loading of MSCs onto proper scaffolds. This achievement extensively supports customized equine tissue engineering to meet various clinical needs of equine bone regeneration in the near future.



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Release the entire work immediately for access worldwide.

Committee Chair

Lopez, Mandi



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