Author

Shane Ruberto

Date of Award

12-2013

Embargo Date

2014

Degree Name

Master of Science (MS)

Department

Biomedical and Chemical Engineering

Advisor(s)

Julie Hasenwinkel

Keywords

Chondroitinase ABC, Chondroitin sulfate proteoglycans, Multivariate regression, Organotypic culture, Raman spectroscopy, Spinal cord injury

Subject Categories

Biochemistry, Biophysics, and Structural Biology | Biomedical Engineering and Bioengineering

Abstract

This study investigated (1) the ability of an organotypic model of spinal cord injury (SCI) to recapitulate significant pathobiological events of in vivo SCI from a Raman spectroscopic perspective, and (2) the Raman spectroscopic effects of chondroitinase ABC (cABC) treatment on said model of SCI. Tissue from mouse spinal cords was excised, sectioned, and placed into culture, before being systematically injured to induce trauma. Cultures with and without cABC treatment were assessed using Raman spectroscopy at varying times post-injury. We demonstrate that Raman spectral phenomena characteristic of SCI can be observed using an organotypic model. By comparing these phenomena with well-known cellular mechanisms of SCI, we hypothesize their correlation with apoptosis, demyelination, and upregulation of endogenous biochemicals, such as chondroitin sulfate proteoglycans (CSPGs), among others. Furthermore, we demonstrate for the first time that Raman spectral phenomena are discernable during cABC treatment of SCI. Observed spectral variations correlate to the enzymatic degradation of CSPGs, protein compositional changes, and remyelination. Multivariate regression modeling confirms changes in lipid and chondroitin sulfate concentration during SCI and its cABC treatment, and provides a semi-quantitative measure of these changes. Overall, these data establish Raman spectroscopy as a viable resource for monitoring cABC treatment of SCI.

Access

Open Access

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