Document Type

Thesis

Date of Degree

Summer 2013

Degree Name

MS (Master of Science)

Degree In

Biomedical Engineering

First Advisor

James A. Martin

Second Advisor

Hongjun Zheng

Abstract

Articular cartilage is an avascular, aneural, and alymphatic tissue with a structure consisting of a superficial, a middle and a deep zone, overlie a calcified zone at the cartilage border between. Each zone has biological and mechanical properties. Self-repair of damaged cartilage seldom if ever occurs, and joint injuries that harm cartilage surfaces often result in osteoarthritis. This has prompted researchers to explore diverse approaches to cartilage regeneration.

The superficial zone shows the highest cellularity and the lowest matrix density. Cartilage cells (chondrocytes) residing in the superficial zone had been thought to be a subpopulation of chondrocytes. However, our laboratory identified a second population of cells that were distinguishable from chondrocytes based on their clonogenicity, multipotency, migratory activity, higher proliferate rate and substantial morphological differences. These cells later proved to be chondrogenic progenitor cells (CPCs). Our continuing studies have shown that CPCs are less chondrogenic than normal chondrocytes and their function is to protect the cartilage surface rather than to regenerate cartilage matrix as previously supposed. In addition, we found evidence to suggest that CPCs act as pro-inflammatory cells in the context of cartilage injury. For these reasons, we undertook a more comprehensive comparison of the phenotypic differences between CPCs and normal chondrocytes and between CPCs and joint cells (tissue synoviocytes from the joint capsule and cells present in synovial fluid) which have been shown to be play roles in joint inflammation.

Gene expression microarray analysis of >25,000 genes revealed that the overall pattern of gene expression in CPCs was distinct from normal chondrocytes, but closely related to synoviocytes and synovial fluid cells. Analysis of specific genes by quantitative PCR (qPCR) showed profound differences between CPCs and normal chondrocytes in terms of cartilage matrix gene expression (Collagen Type ІІ, Aggrecan, Link Protein and COMP) and pro-inflammatory gene expression (IL6, IL8, CCL2 and CXCL12). In contrast, the pattern of CPC gene expression closely resembled. Sulfated glycosaminoglycan assays revealed that cartilage matrix deposition by CPCs, as well as synoviocytes and synovial fluid cells, was significantly inferior to normal chondrocytes. However, chondrogenic and osteogenic differentiation assays, showed no significant differences among the four cell types.

In addition to establishing that CPCs are distinct from chondrocytes, this work suggests significant revisions to our understanding of CPC function in cartilage. The weak chondrogenic ability and higher expression of inflammatory cytokines, suggests these cells don't play a regenerative role as previously thought. On the other, we found evidence that CPCs may form a protective layer on the top of the injured cartilage surfaces, preventing further cartilage injury. In vivo studies are needed to fully elucidate the significance of these roles in cartilage health and disease.

Keywords

cartilage, chondrogenic progenitor cells, microarray, normal chondrocytes, synovial fluid cells, synoviocytes

Pages

x, 57 pages

Bibliography

Includes bibliographical references (pages 54-57).

Copyright

Copyright 2013 Cheng Zhou

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