Abstract
Background. Immunosuppressant therapy is thought to be a major contributor to post-transplant bone disease. Histological data and serum parameters suggest that Cyclosporin A (CsA) treatment causes osteopenia as a result of an altered bone turnover, but the pathogenic mechanisms of this process remain unclear. We investigate if CsA affects cell turnover and extracellular matrix (ECM) synthesis and degradation in MC3T3-E1 osteoblasts, as a surrogate model for in vivo events. Methods. Cell were exposed to increasing doses of CsA (0, 0.5, 1 and 5 μg/ml). Proliferation was evaluated by bromodeoxyuridine (BrdU) incorporation, viability by Trypan Blue exclusion and apoptosis by ELISA. Type I collagen was measured by ELISA and reverse transcription-polymerase chain reaction (RT-PCR), matrix metalloproteinases (MMP) by zymography and RT-PCR, and tissue inhibitors of MMP (TIMP) by reverse zymography. Results. CsA exposure for 48 h decreased osteoblast-number in a dose-dependent manner in the absence of apoptosis or cytotoxicity. CsA at a dose of 5 μg/ml for 72 h caused decreased collagen type I mRNA expression and protein accumulation. While MMP-2 remained unaffected, MMP-9 activity increased. TIMP-1 activity was unaffected, while a dose-dependent increase of TIMP-2 was observed. Conclusions. These data suggest that CsA alters ECM synthesis and degradation in MC3T3-E1 osteoblasts by decreasing type I collagen production and increasing MMP-9 activity. The combination of increased MMP-9 with unchanged TIMP-1 activity could reduce the osteoid matrix available for mineralization. In addition, decreased proliferation could further reduce the number of cells synthesizing new osteoid matrix and thus contribute to the process of bone loss.
| Original language | English |
|---|---|
| Pages (from-to) | 500-505 |
| Number of pages | 6 |
| Journal | Nephrology Dialysis Transplantation |
| Volume | 16 |
| Issue number | 3 |
| State | Published - Apr 2 2001 |
Fingerprint
Keywords
- Cyclosporine
- Extracellular matrix
- Osteoblasts
ASJC Scopus subject areas
- Nephrology
- Transplantation
Cite this
Cyclosporin A affects extracellular matrix synthesis and degradation by mouse MC3T3-E1 osteoblasts in vitro. / Fornoni, Alessia; Cornacchia, Flavia; Howard, Guy; Roos, Bernard A.; Striker, Gary E.; Striker, Liliane J.
In: Nephrology Dialysis Transplantation, Vol. 16, No. 3, 02.04.2001, p. 500-505.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Cyclosporin A affects extracellular matrix synthesis and degradation by mouse MC3T3-E1 osteoblasts in vitro
AU - Fornoni, Alessia
AU - Cornacchia, Flavia
AU - Howard, Guy
AU - Roos, Bernard A.
AU - Striker, Gary E.
AU - Striker, Liliane J.
PY - 2001/4/2
Y1 - 2001/4/2
N2 - Background. Immunosuppressant therapy is thought to be a major contributor to post-transplant bone disease. Histological data and serum parameters suggest that Cyclosporin A (CsA) treatment causes osteopenia as a result of an altered bone turnover, but the pathogenic mechanisms of this process remain unclear. We investigate if CsA affects cell turnover and extracellular matrix (ECM) synthesis and degradation in MC3T3-E1 osteoblasts, as a surrogate model for in vivo events. Methods. Cell were exposed to increasing doses of CsA (0, 0.5, 1 and 5 μg/ml). Proliferation was evaluated by bromodeoxyuridine (BrdU) incorporation, viability by Trypan Blue exclusion and apoptosis by ELISA. Type I collagen was measured by ELISA and reverse transcription-polymerase chain reaction (RT-PCR), matrix metalloproteinases (MMP) by zymography and RT-PCR, and tissue inhibitors of MMP (TIMP) by reverse zymography. Results. CsA exposure for 48 h decreased osteoblast-number in a dose-dependent manner in the absence of apoptosis or cytotoxicity. CsA at a dose of 5 μg/ml for 72 h caused decreased collagen type I mRNA expression and protein accumulation. While MMP-2 remained unaffected, MMP-9 activity increased. TIMP-1 activity was unaffected, while a dose-dependent increase of TIMP-2 was observed. Conclusions. These data suggest that CsA alters ECM synthesis and degradation in MC3T3-E1 osteoblasts by decreasing type I collagen production and increasing MMP-9 activity. The combination of increased MMP-9 with unchanged TIMP-1 activity could reduce the osteoid matrix available for mineralization. In addition, decreased proliferation could further reduce the number of cells synthesizing new osteoid matrix and thus contribute to the process of bone loss.
AB - Background. Immunosuppressant therapy is thought to be a major contributor to post-transplant bone disease. Histological data and serum parameters suggest that Cyclosporin A (CsA) treatment causes osteopenia as a result of an altered bone turnover, but the pathogenic mechanisms of this process remain unclear. We investigate if CsA affects cell turnover and extracellular matrix (ECM) synthesis and degradation in MC3T3-E1 osteoblasts, as a surrogate model for in vivo events. Methods. Cell were exposed to increasing doses of CsA (0, 0.5, 1 and 5 μg/ml). Proliferation was evaluated by bromodeoxyuridine (BrdU) incorporation, viability by Trypan Blue exclusion and apoptosis by ELISA. Type I collagen was measured by ELISA and reverse transcription-polymerase chain reaction (RT-PCR), matrix metalloproteinases (MMP) by zymography and RT-PCR, and tissue inhibitors of MMP (TIMP) by reverse zymography. Results. CsA exposure for 48 h decreased osteoblast-number in a dose-dependent manner in the absence of apoptosis or cytotoxicity. CsA at a dose of 5 μg/ml for 72 h caused decreased collagen type I mRNA expression and protein accumulation. While MMP-2 remained unaffected, MMP-9 activity increased. TIMP-1 activity was unaffected, while a dose-dependent increase of TIMP-2 was observed. Conclusions. These data suggest that CsA alters ECM synthesis and degradation in MC3T3-E1 osteoblasts by decreasing type I collagen production and increasing MMP-9 activity. The combination of increased MMP-9 with unchanged TIMP-1 activity could reduce the osteoid matrix available for mineralization. In addition, decreased proliferation could further reduce the number of cells synthesizing new osteoid matrix and thus contribute to the process of bone loss.
KW - Cyclosporine
KW - Extracellular matrix
KW - Osteoblasts
UR - http://www.scopus.com/inward/record.url?scp=0035095290&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035095290&partnerID=8YFLogxK
M3 - Article
VL - 16
SP - 500
EP - 505
JO - Nephrology Dialysis Transplantation
JF - Nephrology Dialysis Transplantation
SN - 0931-0509
IS - 3
ER -