Due to the fact that bone cells are highly responsive to mechanical stimuli, shear stress has been extensively studied for its ability to enhance osteogenic differentiation through mechanotransduction. In addition, thermal stress has also been explored as a conditioning method to stimulate cellular proliferation, differentiation, and cytoprotection through heat shock protein induction. Despite the beneficial effects observed with individual stress on cells, there has been little focus on the potential of a combination of stresses to improve cellular response. Therefore, the aim of this study was to investigate the effect of combined shear and thermal stress on preosteoblasts to stimulate an enhanced osteogenic response. To achieve this, MC3T3-E1 cells were exposed to one of the following protocols for an hour: no stress (control), shear stress at 1 dyne/cm2 using a parallel plate flow chamber, thermal stress in a 42°C incubator, or combined shear and thermal stress (1 dyne/cm2 at 42°C). Stress treatments were applied on Day 2, Day 6, and Day 10. To assess the early response of cells to stress treatments, we measured metabolic activity, expression of signaling factors, and HSPs following stress on Day 2. Despite an initial decrease in metabolism, combined stress stimulated a strong response in VEGF (12.49 RFI) COX-2 (12.32 RFI), HSPs (2-4 RFI) and increased PGE accumulation. The long-term cellular response to stress treatments was measured on Day 15 by evaluating the ability of combined stress to stimulate late stage markers of differentiation. Combined stress increased OPN gene and protein expression, yet OCN was minimally affected by stress treatments. However, mineralization was significantly decreased with combined stress. Overall, combined stress was able to stimulate an enhanced effect across a majority of the bone-related markers measured, whereas individual shear stress or thermal stress were limited in their response. This suggests that combined stress can provide the appropriate cues to modify osteoblast differentiation and generate an enhanced osteogenic response. / Master of Science
| Identifer | oai:union.ndltd.org:VTETD/oai:vtechworks.lib.vt.edu:10919/50822 |
| Date | 06 November 2014 |
| Creators | Sampson, Alana Cherrell |
| Contributors | Biomedical Engineering, Rylander, M. Nichole, Goldstein, Aaron S., Freeman, Joseph W. |
| Publisher | Virginia Tech |
| Source Sets | Virginia Tech Theses and Dissertation |
| Detected Language | English |
| Type | Thesis |
| Format | ETD, application/pdf |
| Rights | In Copyright, http://rightsstatements.org/vocab/InC/1.0/ |
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