Date of Award

Fall 12-8-2023

Degree Type

Thesis

Degree Name

MS Biology

Department

Biology

Advisor

Jessica Cottrell, PhD

Committee Member

Daniel Brian Nicholas, PhD

Committee Member

Constantine Bitsaktsis, PhD

Keywords

Inflammation, Chondrocytes, ROS/RNS, Cytokine Expression, CRP, Copper (ii) Chloride

Abstract

Copper (II) chloride is a common ionic salt that is present in the environment as well as various kinds of pesticides used in the agricultural industry. Previous research has shown that this molecule increases inflammation in ATDC5 chondrocytes, during the process of chondrogenesis. However, the mechanism by which copper (II) chloride induces inflammation is not well understood. As such, chondrocytes were cultured and examined at different timepoints (i.e., days 0, 1, 3, 7, 10, and 14) to better understand the induction of inflammation. The presence/absence of inflammation in the chondrocytes was determined through various tests such as an ROS/RNS quantification, qPCR gene expression analysis of different proinflammatory cytokines, and measuring C Reactive protein levels. Levels of superoxides and nitric oxide significantly increased by day 7 in a dose-dependent manner. Furthermore, the tested aggrecanase ADAMTS5 and the cytokines IL-17α, IL-18, and IL-6 were elevated at different points along in the timeline. ADAMTS5 and IL-17α were elevated on day 1 of the experiment, followed by an increase in IL-6 and IL-18 that peaked on day 3. Days 7 and 10 showed a decline in all cytokines other than IL-17α, which increased significantly in expression. CRP levels reflected a dose-dependent trend similar to the results of the ROS/RNS assay. Our data indicates CuCl2 induces inflammation in the ATDC5 chondrocytes primarily through the release of pro-inflammatory cytokines along with the concomitant accumulation of ROS/RNS species.

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