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Effects of crystalline glucocorticoid triamcinolone acetonide on cultered human supraspinatus tendon cells.

Tempfer H, Gehwolf R, Lehner C, Wagner A, Mtsariashvili M, Bauer HC, Resch H, Tauber M - Acta Orthop (2009)

Bottom Line: Moreover, expression and secretion of collagen I was strongly reduced, and there was a decrease in proliferation rate.Cell migration was blocked and the rate of expression of the matrix metalloproteinases MMP2, MMP8, MMP9, and MMP13 was reduced, but expression of TIMP1 (a tissue inhibitor of MMPs) was upregulated, indicating a reduction in the cellular capacity for tendon repair.These results may indicate that the use of TAA is one reason for weaker mechanical tendon properties and for the high rate of re-rupture after supraspinatus tendon repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Organismic Biology, Division of Zoology and Functional Anatomy, University of Salzburg, Austria. Herbert.Tempfer@sbg.ac.at

ABSTRACT

Background: Rotator cuff tears are a common cause of shoulder pain and impairment. Subacromial glucocorticoid injections are widely used for treatment of epiphenomenons of chronic impingement syndrome with the possible side effects of tendon rupture and impaired tendon healing.

Methods: Using qRT-PCR, western blot, immunoflourescence, and measurement of 3H-thymidine uptake we investigated the effects of the crystalline glucocorticoid triamcinolone acetonide (TAA) when added to the culture medium of isolated human rotator cuff tendon cells.

Results: After 2 weeks of incubation, the cells had lost their fibroblastic appearance and parallel orientation, which is characteristic of cellular degeneration in vivo. Moreover, expression and secretion of collagen I was strongly reduced, and there was a decrease in proliferation rate. Cell migration was blocked and the rate of expression of the matrix metalloproteinases MMP2, MMP8, MMP9, and MMP13 was reduced, but expression of TIMP1 (a tissue inhibitor of MMPs) was upregulated, indicating a reduction in the cellular capacity for tendon repair. In addition, changes in cellular differentiation were observed: the number of adipocytes increased and levels of the protein Sox9-a marker of differentiating and mature chondrocytes-were elevated in triamcinolone acetonide treated cells.

Interpretation: These results may indicate that the use of TAA is one reason for weaker mechanical tendon properties and for the high rate of re-rupture after supraspinatus tendon repair.

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Related in: MedlinePlus

A. Results of gelatin zymography showing the downregulation of MMP2 and MMP9 by triamcinolone acetonide (right lane) compared to untreated control cells (left lane). RT-PCR result showing downregulation of MMP2, MMP8, MMP9, and MMP13 mRNA levels after TAA treatment. B. Levels of MMP1, MMP3, and MMP14 mRNA were unchanged. C. qRT-PCR showed significant downregulation of MMP2 and MMP9 mRNA levels, and significant upregulation of TIMP1 mRNA. Gene expression levels of MMP2, MMP9, and TIMP1 were normalized to the expression of housekeeping genes GAPDH and HPRT. * p < 0.001.
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Figure 0003: A. Results of gelatin zymography showing the downregulation of MMP2 and MMP9 by triamcinolone acetonide (right lane) compared to untreated control cells (left lane). RT-PCR result showing downregulation of MMP2, MMP8, MMP9, and MMP13 mRNA levels after TAA treatment. B. Levels of MMP1, MMP3, and MMP14 mRNA were unchanged. C. qRT-PCR showed significant downregulation of MMP2 and MMP9 mRNA levels, and significant upregulation of TIMP1 mRNA. Gene expression levels of MMP2, MMP9, and TIMP1 were normalized to the expression of housekeeping genes GAPDH and HPRT. * p < 0.001.

Mentions: In qRT-PCR applications, expression of MMP2 and MMP9 mRNA was found to be significantly downregulated by TAA treatment—by 58% (SD 0.9) for MMP2 and by 72% (SD 1.9) for MMP9—whereas the expression of TIMP1 mRNA was upregulated by 111% (SD 21). The decrease in MMP2 and MMP9 levels was confirmed by gelatine zymography. In the case of MMP9, all the functional forms of the enzyme that have been described to date (active MMP9 with a size of 92 kD, pro-MMP9 of about 130 kD, and the homodimer of 225 kD) were downregulated (Figure 3).


Effects of crystalline glucocorticoid triamcinolone acetonide on cultered human supraspinatus tendon cells.

Tempfer H, Gehwolf R, Lehner C, Wagner A, Mtsariashvili M, Bauer HC, Resch H, Tauber M - Acta Orthop (2009)

A. Results of gelatin zymography showing the downregulation of MMP2 and MMP9 by triamcinolone acetonide (right lane) compared to untreated control cells (left lane). RT-PCR result showing downregulation of MMP2, MMP8, MMP9, and MMP13 mRNA levels after TAA treatment. B. Levels of MMP1, MMP3, and MMP14 mRNA were unchanged. C. qRT-PCR showed significant downregulation of MMP2 and MMP9 mRNA levels, and significant upregulation of TIMP1 mRNA. Gene expression levels of MMP2, MMP9, and TIMP1 were normalized to the expression of housekeeping genes GAPDH and HPRT. * p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2823208&req=5

Figure 0003: A. Results of gelatin zymography showing the downregulation of MMP2 and MMP9 by triamcinolone acetonide (right lane) compared to untreated control cells (left lane). RT-PCR result showing downregulation of MMP2, MMP8, MMP9, and MMP13 mRNA levels after TAA treatment. B. Levels of MMP1, MMP3, and MMP14 mRNA were unchanged. C. qRT-PCR showed significant downregulation of MMP2 and MMP9 mRNA levels, and significant upregulation of TIMP1 mRNA. Gene expression levels of MMP2, MMP9, and TIMP1 were normalized to the expression of housekeeping genes GAPDH and HPRT. * p < 0.001.
Mentions: In qRT-PCR applications, expression of MMP2 and MMP9 mRNA was found to be significantly downregulated by TAA treatment—by 58% (SD 0.9) for MMP2 and by 72% (SD 1.9) for MMP9—whereas the expression of TIMP1 mRNA was upregulated by 111% (SD 21). The decrease in MMP2 and MMP9 levels was confirmed by gelatine zymography. In the case of MMP9, all the functional forms of the enzyme that have been described to date (active MMP9 with a size of 92 kD, pro-MMP9 of about 130 kD, and the homodimer of 225 kD) were downregulated (Figure 3).

Bottom Line: Moreover, expression and secretion of collagen I was strongly reduced, and there was a decrease in proliferation rate.Cell migration was blocked and the rate of expression of the matrix metalloproteinases MMP2, MMP8, MMP9, and MMP13 was reduced, but expression of TIMP1 (a tissue inhibitor of MMPs) was upregulated, indicating a reduction in the cellular capacity for tendon repair.These results may indicate that the use of TAA is one reason for weaker mechanical tendon properties and for the high rate of re-rupture after supraspinatus tendon repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Organismic Biology, Division of Zoology and Functional Anatomy, University of Salzburg, Austria. Herbert.Tempfer@sbg.ac.at

ABSTRACT

Background: Rotator cuff tears are a common cause of shoulder pain and impairment. Subacromial glucocorticoid injections are widely used for treatment of epiphenomenons of chronic impingement syndrome with the possible side effects of tendon rupture and impaired tendon healing.

Methods: Using qRT-PCR, western blot, immunoflourescence, and measurement of 3H-thymidine uptake we investigated the effects of the crystalline glucocorticoid triamcinolone acetonide (TAA) when added to the culture medium of isolated human rotator cuff tendon cells.

Results: After 2 weeks of incubation, the cells had lost their fibroblastic appearance and parallel orientation, which is characteristic of cellular degeneration in vivo. Moreover, expression and secretion of collagen I was strongly reduced, and there was a decrease in proliferation rate. Cell migration was blocked and the rate of expression of the matrix metalloproteinases MMP2, MMP8, MMP9, and MMP13 was reduced, but expression of TIMP1 (a tissue inhibitor of MMPs) was upregulated, indicating a reduction in the cellular capacity for tendon repair. In addition, changes in cellular differentiation were observed: the number of adipocytes increased and levels of the protein Sox9-a marker of differentiating and mature chondrocytes-were elevated in triamcinolone acetonide treated cells.

Interpretation: These results may indicate that the use of TAA is one reason for weaker mechanical tendon properties and for the high rate of re-rupture after supraspinatus tendon repair.

Show MeSH
Related in: MedlinePlus