Publication Date

1991

Document Type

Dissertation/Thesis

First Advisor

Johnson-Wint, Barbara

Degree Name

M.S. (Master of Science)

Department

Department of Biological Sciences

LCSH

Collagen||Colloids||Serum||Proteinase

Abstract

Rotating and stationary floating collagen type I gels, seeded with rabbit corneal fibroblasts, were incubated in the presence or absence of calf serum, whole soluble plasma fibronectin, 29 kDa amino terminal fibronectin fragment, Pronectin F, or ultrogel AcA 54 fractionated calf serum. The corneal stromal cells were examined for collagen gel contraction by direct measurement of gel area and matrix metalloproteinase production using sodium dodecyl sulfate (SDS) - polyacrylamide gel zymography of their culture medium. Nonrotating collagen gels showed increased contraction of the collagen gels and no detectable production of MMP-1 and MMP-3 with increasing concentrations of calf serum, whereas collagen gels with no serum showed much less contraction and detected MMP-1 and MMP-3 in the culture medium. Detectable levels of MMP-2 and MMP-9 remained constituitive and were unaffected by serum or a lack of serum. Fibronectin, Pronectin F (bioengineered RGDS repeated protein polymer), and 29 kDa amino terminal fibronectin fragment added into cultures containing no serum did not replace effects of serum on gel contraction or MMP-1 and MMP-3 production. Detectable levels of MMP-2 and MMP-9 remained constituitive and were unaffected by adding fibronectin, Pronectin F, and 29 kDa amino terminal fibronectin fragment. To determine which factor(s) are involved in serum mediated collagen contraction and suppression of MMP production, calf serum was fractionated on a gel chromatography column. Higher molecular weight calf serum fractions resulted in improved contraction of collagen gels and showed a marked reduction in MMP-1 and MMP-3 production while MMP-2 and MMP-9 levels remained constituitive. Continuous rotation of the collagen gel cultures enhanced collagen gel contraction and MMP-1 and MMP-3 production with respect to previously nonrotated cultures. In summary, rotating corneal fibroblast seeded collagen type I gel cultures suggest that a high molecular weight protein in serum, not fibronectin, allows proper attachment and contraction of the collagen, prevents matrix metalloproteinase-1 and -3 production, and does not affect constituitive production of MMP-2 and MMP-9.

Comments

Includes bibliographical references (pages [62]-70).

Extent

70 pages

Language

eng

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

Media Type

Text

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