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SPECTROSCOPIC ASSESSMENT OF TISSUE ENGINEERED CARTILAGE: A PATHWAY FOR BENCH TO BEDSIDE EVALUATION OF CARTILAGE DEVELOPMENT AND REPAIR
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Thesis/Dissertation
Date
2021
Advisor
Pleshko, Nancy
Committee member
Patil, Chetan Appasaheb
Orrego, Santiago
Mauck, Robert L.
Orrego, Santiago
Mauck, Robert L.
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Department
Bioengineering
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DOI
http://dx.doi.org/10.34944/dspace/7625
Abstract
Cartilage tissue engineering is a promising approach for the repair of chondral defects. Engineering of cartilage combines a three-dimensional scaffold with chondrogenic cells and appropriate external stimuli, and ideally results in constructs with properties that resemble, as closely as possible, native cartilage. Once implanted to repair a cartilage defect, the integration of tissue engineered cartilage (TEC) to surrounding native tissue is critical for a successful clinical outcome. However, this depends in part on the initial maturity of the engineered construct, which is challenging to assess a priori. Another challenge relates to the assessment of the longitudinal repair of cartilage defects after tissue engineering approaches are applied. Currently, evaluation is qualitative, based on visual and tactile observations using arthroscopic hook probes which can be a very subjective approach. Furthermore, gold standard techniques (histological, mechanical and biochemical evaluation) to determine compositional and mechanical properties of constructs in vitro and ex vivo are generally destructive.In this thesis, we are proposing the use of a spectroscopic fiber optic probe approach that spans the visible-near infrared (Vis-NIR) regions. This would be a novel, non-destructive technique based on high frequency nonionizing radiation that causes vibrations in the NIR region (750-2500 nm or 12000 to 4000 cm-1), and electronic transitions in the VIS region (400-750 nm) that result in a unique spectrum of the sampled tissues. Previously, we have shown that NIR spectral data collected in a non-destructive manner correlate to compositional and biomechanical properties of tissue engineered cartilage. Additionally, using an arthroscopic fiber optic probe, Vis-NIR spectra can be collected from repairing cartilage tissue in situ.
The overarching hypothesis of this thesis is that Vis-NIR fiber optic spectroscopy can be utilized to assess engineered cartilage development in vitro, and in vivo to assess repair in a pre-clinical model of chondral defect. This hypothesis was tested in the following three aims: 1. Assessment of the mid and NIR spectral features of scaffold and extracellular matrix components of cartilage and TECs for in vitro monitoring of TEC development by fiber optic NIR spectroscopy; 2. Assessment of the Vis-NIR spectral features of tissues present in the mini-pig stifle joint during the chondral repair process to facilitate interpretation of in vivo repair; 3. Model-informed design and analysis of an arthroscopic probe for spectral collection during the cartilage repair process in preclinical models and clinical scenarios. Together these studies contribute to an overall approach for spectroscopic assessment of tissue engineered cartilage as a pathway for bench to bedside evaluation of cartilage development and repair.
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