MMP-2 mediates local degradation and remodeling of collagen by annulus fibrosus cells of the intervertebral disc

dc.contributor.authorRastogi, Anshu
dc.contributor.authorKim, Hyunchul
dc.contributor.authorTwomey, Julianne D
dc.contributor.authorHsieh, Adam H
dc.date.accessioned2021-09-27T19:57:46Z
dc.date.available2021-09-27T19:57:46Z
dc.date.issued2013-04-27
dc.description.abstractDegeneration of the intervertebral disc (IVD) is characterized by marked degradation and restructuring of the annulus fibrosus (AF). Although several matrix metalloproteinases (MMPs) have been found to be more prevalent in degenerate discs, their coordination and function within the context of the disease process are still not well understood. In this study, we sought to determine whether MMP-2 is associated with degenerative changes in the AF and to identify the manner by which AF cells use MMP-2. Two established animal models of disc degeneration, static compression and transannular needle puncture of rodent caudal discs, were examined for MMP-2 immunopositivity. With lentiviral transduction of an shRNA expression cassette, we screened and identified an effective shRNA sequence for generating stable RNA interference to silence MMP-2 expression in primary rat AF cells. Gelatin films were used to compare gelatinase activity and spatial patterns of degradation between transduced cells, and both noninfected and nonsense shRNA controls. The functional significance of MMP-2 was determined by assessing the ability for cells to remodel collagen gels. Both static compression and 18-g annular puncture of rodent caudal discs stimulated an increase in MMP-2 activity with concurrent lamellar disorganization in the AF, whereas 22-g and 26-g needle injuries did not. To investigate the functional role of MMP-2, we established lentivirus-mediated RNAi to induce stable knockdown of transcript levels by as much as 88%, and protein levels by as much as 95% over a 10-day period. Culturing transduced cells on gelatin films confirmed that MMP-2 is the primary functional gelatinase in AF cells, and that MMP-2 is used locally in regions immediately around AF cells. In collagen gels, transduced cells demonstrated an inability to remodel collagen matrices. Our study indicates that increases in MMP-2 observed in human degenerate discs are mirrored in experimentally induced degenerative changes in rodent animal models. AF cells appear to use MMP-2 in a very directed fashion for local matrix degradation and collagen remodeling. This suggests that MMP-2 may have a functionally significant role in the etiology of degenerative disc disease and could be a potential therapeutic target.en_US
dc.description.urihttps://doi.org/10.1186/ar4224
dc.description.urihttps://doi.org/10.1186/1471-2164-14-287
dc.identifierhttps://doi.org/10.13016/g4gk-hsji
dc.identifier.citationRastogi, A., Kim, H., Twomey, J.D. et al. MMP-2 mediates local degradation and remodeling of collagen by annulus fibrosus cells of the intervertebral disc. Arthritis Res Ther 15, R57 (2013).en_US
dc.identifier.urihttp://hdl.handle.net/1903/28028
dc.language.isoen_USen_US
dc.publisherSpringer Natureen_US
dc.relation.isAvailableAtA. James Clark School of Engineeringen_us
dc.relation.isAvailableAtFischell Department of Bioengineeringen_us
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_us
dc.relation.isAvailableAtUniversity of Maryland (College Park, MD)en_us
dc.subjectIntervertebral discen_US
dc.subjectIntervertebral discen_US
dc.subjectCollagenen_US
dc.subjectRemodelingen_US
dc.subjectRNA interferenceen_US
dc.titleMMP-2 mediates local degradation and remodeling of collagen by annulus fibrosus cells of the intervertebral discen_US
dc.typeArticleen_US

Files

Original bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
ar4224.pdf
Size:
5.75 MB
Format:
Adobe Portable Document Format
Description:
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.57 KB
Format:
Item-specific license agreed upon to submission
Description: