Dissertation
ENGINEERING CELL CULTURE PLATFORMS USING MECHANICAL STRAIN, PERFUSION, AND HYPOXIA FOR CARTILAGE TISSUE ENGINEERING APPLICATIONS
Washington State University
Doctor of Philosophy (PhD), Washington State University
01/2021
DOI:
https://doi.org/10.7273/000005511
Handle:
https://hdl.handle.net/2376/119082
Abstract
Articular Cartilage (AC) is an avascular protective soft tissue that covers bones in articulating joints. The avascular nature of AC makes its access to oxygen and nutrients limited and reduces its ability to regenerate after trauma. AC degenerates gradually after injury leading to osteoarthritis (OA), the most common form of arthritis worldwide. Unfortunately, there is no cure for OA, and conventional treatments, including total knee arthroscopy (TKA) and microfracture, have failed to restore the functionality of cartilage. Articular cartilage tissue engineering (ACTE) is being developed to provide an alternative approach for tackling OA. The success of ACTE relies heavily on the availability of chondrogenic cell lines, using biochemical cues that improve the ability of cells to secrete a robust extracellular matrix (ECM), the development of dynamic bioreactors that stimulate ECM production and creating platforms that mimic the native environment in the knee. This dissertation is focused on engineering cell culture platforms that can be potentially used for enhancing lab-grown AC tissue to improve ACTE success. First, an optimum co-culture ratio of articular chondrocytes (ACh) and adipose-derived stem cells (ASCs) has been defined as a cell source for autologous chondrocytes implantation (ACI). Second, a unique approach for expanding and preparing chondrocyte, that combines dynamic stretching and gallic acid supplementation, was developed for better ACI outcomes. Third, a novel perfusion system was developed to enhance the ECM production by diseased AChs encapsulated in porous scaffolds. The beauty of this combination, perfusion, and porous scaffolds, is that a higher shear pressure is observed within the scaffold to stimulate AC growth, even when low rates of medium flow are used. Finally, a low-cost hypoxia chamber was designed, assembled, and used to simulate viability, DNA, and ECM production by chondrocytes encapsulated in agarose scaffolds. This system may be simply replicated in any laboratory to utilize the benefits of hypoxia for cartilage tissue engineering.
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Details
- Title
- ENGINEERING CELL CULTURE PLATFORMS USING MECHANICAL STRAIN, PERFUSION, AND HYPOXIA FOR CARTILAGE TISSUE ENGINEERING APPLICATIONS
- Creators
- Haneen A Abusharkh
- Contributors
- Bernard J Van Wie (Advisor)Nehal I Abu-Lail (Committee Member)Bulent A Gozen (Committee Member)Alla Kostyukova (Committee Member)David B Thiessen (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Voiland College of Engineering and Architecture
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Publisher
- Washington State University
- Number of pages
- 142
- Identifiers
- 99900592259801842
- Language
- English
- Resource Type
- Dissertation