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Progression of chondrocyte signaling responses to mechanical stimulation in 3-D gel culture

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dc.contributor.advisor Alan J. Grodzinsky. en_US
dc.contributor.author Chai, Diana H en_US
dc.contributor.other Massachusetts Institute of Technology. Biological Engineering Division. en_US
dc.date.accessioned 2008-09-02T17:58:34Z
dc.date.available 2008-09-02T17:58:34Z
dc.date.copyright 2008 en_US
dc.date.issued 2008 en_US
dc.identifier.uri http://hdl.handle.net/1721.1/42071
dc.description Thesis (Ph. D.)--Massachusetts Institute of Technology, Biological Engineering Division, 2008. en_US
dc.description This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections. en_US
dc.description Includes bibliographical references (leaves 148-156). en_US
dc.description.abstract Mechanical stimulation of 3-D chondrocyte cultures increases extracellular matrix (ECM) production and mechanical stiffness in regenerating cartilage. The goal of this study was to examine the progression of chondrocyte signaling responses to mechanical stimulation in 3-D culture during tissue regeneration. To investigate the role of integrins in chondrocyte mechanotransduction, function-blocking antibodies and small-molecule antagonists were used to disrupt integrin-matrix interactions during dynamic compression of chondrocytes in 3-D agarose culture. At early days in culture, blocking [alpha]v[beta]3 integrin abolished dynamic compression stimulation of proteoglycan synthesis, independent of effects in free-swell culture, while blocking [alpha]5[beta]1 integrins abolished the effect of compression only when blocking in free-swell increased proteoglycan synthesis. This suggests that disrupting [alpha]v[beta]3 and [alpha]5[beta]1 interactions with the ECM influences proteoglycan synthesis in distinct pathways and that [alpha]v[beta]3 more directly influences the mechanical response. To further distinguish individual mechanotransduction pathways, we investigated the temporal gene transcription response of chondrocytes to ramp-and-hold compression on Days 1, 10, and 28 in 3-D agarose culture. Clustered and individual gene expression profiles changed temporally and in magnitude over time in culture. Day 1 cultures differed from Days 10 and 28, reflecting changes in cell microenvironment with development of pericellular and extracellular matrices. Comparisons with the response of intact tissue to compression suggested similar regulatory mechanisms. We further investigated MAPkinase (ERK1/2, p38, JNK) and Akt activation on Days 1 and 28 in agarose culture through phosphorylation state-specific Western blotting. en_US
dc.description.abstract (cont.) Compression induced transient ERK1/2 phosphorylation on both days, with Day 28 levels similar to intact tissue. Unique from tissue behavior, only slight transient p38 phosphorylation was observed on Day 28, and SEK phosphorylation was undetected. Akt was uniquely regulated in intact cartilage compared to MAPks, with decreased total Akt levels over time under static compression. In contrast, compression transiently decreased pAkt levels in agarose cultures, with no changes in total Akt. Changes in the chondrocyte responses to compression with time in agarose culture suggest that cells sense different forces and respond differently with time; further studies may help optimize mechanical loading for tissue-engineering purposes. These studies provide a basis for further examination of mechanotransduction in cartilage. en_US
dc.description.statementofresponsibility by Diana H. Chai. en_US
dc.format.extent 156 leaves en_US
dc.language.iso eng en_US
dc.publisher Massachusetts Institute of Technology en_US
dc.rights M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. en_US
dc.rights.uri http://dspace.mit.edu/handle/1721.1/7582 en_US
dc.subject Biological Engineering Division. en_US
dc.title Progression of chondrocyte signaling responses to mechanical stimulation in 3-D gel culture en_US
dc.type Thesis en_US
dc.description.degree Ph.D. en_US
dc.contributor.department Massachusetts Institute of Technology. Biological Engineering Division. en_US
dc.identifier.oclc 234531899 en_US


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