Publication Date

2013-06-17

Availability

Open access

Embargo Period

2013-06-17

Degree Name

Master of Science (MS)

Department

Biomedical Engineering (Engineering)

Date of Defense

2013-05-16

First Committee Member

Chun-Yuh Charles Huang

Second Committee Member

Alicia R. Jackson

Third Committee Member

Weiyong Gu

Abstract

Lower back pain (LBP) and cartilage-related diseases such as osteoarthritis (OA) have a tremendous financial impact on the world today. It is well-understood that the provision of glucose to these avascular regions of the body plays a large role in the sustenance of health and the prevention of pathogenic processes. In order to undertake innovative approaches to allow for glucose-involved cell therapeutics and to enhance a wide range of stem cell therapies, we need to better understand avascular cell metabolism. In this study, annulus fibrosus (AF), nucleus pulposus (NP), and cartilage cells were placed in agarose gel constructs and incubated in varying concentrations of glucose for up to 12 days. Live cell density was quantified using a viability staining technique at varying time points. The stained samples were imaged and cropped using a standard area, and the cells in each image were counted using ImageJ. Cell density results indicated that cell viability can be maintained at 0.5 mM glucose for all cell types. NP cells appeared to lose viability when incubated in high concentrations of glucose medium. There was a statistically significant drop in cell density of NP cells incubated in 3.2 g/L vs. those in 0.1 g/L glucose medium. Glucose consumption was analyzed using a glucose assay kit in conjunction with absorbance readings from a spectrophotometer. AF cells consumed the most glucose in the first 24 hours of culture, followed by NP cells and then chondrocytes. The amount of glucose consumed increased for all cell types when cultured in higher concentrations of glucose.

Keywords

glucose; chondrocytes; ivd; Intervertebral disc degeneration; avascular; cell metabolism

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