Investigation of the Effect of Trace Element, Copper Chloride, on Chondrogenic Cell Line, ATDC5 Function

Author

Alexandra BambrickFollow

Date of Award

Fall 12-20-2022

Degree Type

Thesis

Degree Name

MS Microbiology

Department

Biology

Advisor

Jessica Cottrell, Ph.D.

Committee Member

Jessica Cottrell, Ph.D.

Committee Member

Daniel Brian Nichols, Ph.D.

Committee Member

Constantine Bitsaktsis, Ph.D.

Keywords

Chondrocyte, Cartilage, Chondrogenesis, Osteoarthritis, Copper chloride, Cell Culture

Abstract

Copper chloride (CuCl₂) is an essential trace element found in humans, which helps to aid in brain and nerve function, glucose metabolism, calcium absorption, and bone formation. Bone formation utilizes a variety of cell types, including, osteoblasts, osteoclasts, and chondrocytes. Chondrocytes deposit cartilage components like collagen and proteoglycans that provide the necessary scaffold for osteoblasts to synthesize and mineralize bone. Copper chloride is known to aid in osteogenesis, maintain bone metabolism, and regulate bone mineral density, but its role in chondrogenesis is still unclear (15).

In this study, it is hypothesized that CuCl₂ treatments would enhance chondrogenic differentiation in the murine chondrogenic ATDC5 cell line. To test this hypothesis, chondrocytes were treated with a no treatment control (DMEM/F12) (Corning, Corning NY), a positive control of insulin (2.8mM), and increasing doses CuCl₂ (2.6mM, 26mM, 260mM) over 14 days. Cell media was collected 18-20 hours post-treatment on days 0, 1, 3, 7, 10, and 14. Additional experiments were run with spent media to observe the effect of copper chloride on functionality. The amount of alkaline phosphatase (ALP) activity, an important biomarker of chondrocyte activity was measured overtime throughout treatments. At 14 days post-treatment, alizarin red staining was performed to quantify calcium deposited in the presence of CuCl₂ treated cells. To observe gene expression changes, quantitative polymerase chain reaction (qPCR) was run for each sample. Specific genes to chondrocytes were run including, VEGF, aggrecan, COL2A1, COL2B1, KISS1, and RHEB. Results found that CuCl₂ decreased cell proliferation and ALP activity at medium and high doses. Calcium deposition as well as proteoglycan activity both increased as doses of CuCl₂ increased. Overall, by understanding these effects on chondrogenesis advances our knowledge on the environmental impact they have on organismal health.

Recommended Citation

Bambrick, Alexandra, "Investigation of the Effect of Trace Element, Copper Chloride, on Chondrogenic Cell Line, ATDC5 Function" (2022). Seton Hall University Dissertations and Theses (ETDs). 3045.
https://scholarship.shu.edu/dissertations/3045