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Decreased CD90 expression in human mesenchymal stem cells by applying mechanical stimulation.

Wiesmann A, Bühring HJ, Mentrup C, Wiesmann HP - Head Face Med (2006)

Bottom Line: In stimulated and unstimulated cultures an increase in the calcium content over time was observed.These results demonstrate the influence of mechanical force on the differentiation of human MSC into osteoblast-like cells in vitro.While significant enhancement of the biomineral formation by mechanical stimulation is not detected before 21 days, effects on the extracellular matrix became already obvious after 14 days.

View Article: PubMed Central - HTML - PubMed

Affiliation: Medizinische Klinik und Poliklinik, Abteilung für Hämatologie, Onkologie, Immunologie und Rheumatologie, Universitätsklinikum Tübingen, Tübingen, Germany. anne.wiesmann@med.uni-tuebingen.de

ABSTRACT

Background: Mesenchymal stem cells (MSC) are multipotent cells which can differentiate along osteogenic, chondrogenic, and adipogenic lineages. The present study was designed to investigate the influence of mechanical force as a specific physiological stress on the differentiation of (MSC) to osteoblast-like cells.

Methods: Human MSC were cultured in osteoinductive medium with or without cyclic uniaxial mechanical stimulation (2000 mustrain, 200 cycles per day, 1 Hz). Cultured cells were analysed for expression of collagen type I, osteocalcin, osteonectin, and CD90. To evaluate the biomineral formation the content of bound calcium in the cultures was determined.

Results: After 14 days in culture immunfluorescence staining revealed enhancement of collagen type I and osteonectin expression in response to mechanical stimulation. In contrast, mechanically stimulated cultures stained negative for CD90. In stimulated and unstimulated cultures an increase in the calcium content over time was observed. After 21 days in culture the calcium content in mechanical stimulated cultures was significantly higher compared to unstimulated control cultures.

Conclusion: These results demonstrate the influence of mechanical force on the differentiation of human MSC into osteoblast-like cells in vitro. While significant enhancement of the biomineral formation by mechanical stimulation is not detected before 21 days, effects on the extracellular matrix became already obvious after 14 days. The decrease of CD90 expression in mechanically stimulated cultures compared to unstimulated control cultures suggests that CD90 is only transiently expressed expression during the differentiation of MSC to osteoblast-like cells in culture.

No MeSH data available.


Related in: MedlinePlus

Concentration of bound calcium in human MCS cultures. Grey bars correspond to cultures cultivated in osteoinduction medium containing dexamethasone, ascorbate and β-glycerophosphate with uniaxial mechanical stimulation (2000 μstrain, 200 × day, 1 Hz). White bars correspond to control cultures without mechanical stimulation. The calcium concentration was measured at day 7, 14 and 21. Statistically significant differences between mechanically stimulated and mechanically unstimulated cultures are indicated by *.
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Figure 3: Concentration of bound calcium in human MCS cultures. Grey bars correspond to cultures cultivated in osteoinduction medium containing dexamethasone, ascorbate and β-glycerophosphate with uniaxial mechanical stimulation (2000 μstrain, 200 × day, 1 Hz). White bars correspond to control cultures without mechanical stimulation. The calcium concentration was measured at day 7, 14 and 21. Statistically significant differences between mechanically stimulated and mechanically unstimulated cultures are indicated by *.

Mentions: The calcium concentrations in the mechanically stimulated and unstimulated cultures were determined after 7, 14 and 21 days in culture. The results are presented in Fig. 3. For stimulated and unstimulated cultures an increase in the calcium concentration could be observed. This increase is more prominent for the mechanically stimulated cultures. After 7 days of culture there is no difference in calcium concentration between mechanically unstimulated and mechanically stimulated cultures. In contrast, after 3 weeks in culture, there is a significant higher calcium concentration in mechanically stimulated cultures compared to mechanically unstimulated cultures.


Decreased CD90 expression in human mesenchymal stem cells by applying mechanical stimulation.

Wiesmann A, Bühring HJ, Mentrup C, Wiesmann HP - Head Face Med (2006)

Concentration of bound calcium in human MCS cultures. Grey bars correspond to cultures cultivated in osteoinduction medium containing dexamethasone, ascorbate and β-glycerophosphate with uniaxial mechanical stimulation (2000 μstrain, 200 × day, 1 Hz). White bars correspond to control cultures without mechanical stimulation. The calcium concentration was measured at day 7, 14 and 21. Statistically significant differences between mechanically stimulated and mechanically unstimulated cultures are indicated by *.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1483821&req=5

Figure 3: Concentration of bound calcium in human MCS cultures. Grey bars correspond to cultures cultivated in osteoinduction medium containing dexamethasone, ascorbate and β-glycerophosphate with uniaxial mechanical stimulation (2000 μstrain, 200 × day, 1 Hz). White bars correspond to control cultures without mechanical stimulation. The calcium concentration was measured at day 7, 14 and 21. Statistically significant differences between mechanically stimulated and mechanically unstimulated cultures are indicated by *.
Mentions: The calcium concentrations in the mechanically stimulated and unstimulated cultures were determined after 7, 14 and 21 days in culture. The results are presented in Fig. 3. For stimulated and unstimulated cultures an increase in the calcium concentration could be observed. This increase is more prominent for the mechanically stimulated cultures. After 7 days of culture there is no difference in calcium concentration between mechanically unstimulated and mechanically stimulated cultures. In contrast, after 3 weeks in culture, there is a significant higher calcium concentration in mechanically stimulated cultures compared to mechanically unstimulated cultures.

Bottom Line: In stimulated and unstimulated cultures an increase in the calcium content over time was observed.These results demonstrate the influence of mechanical force on the differentiation of human MSC into osteoblast-like cells in vitro.While significant enhancement of the biomineral formation by mechanical stimulation is not detected before 21 days, effects on the extracellular matrix became already obvious after 14 days.

View Article: PubMed Central - HTML - PubMed

Affiliation: Medizinische Klinik und Poliklinik, Abteilung für Hämatologie, Onkologie, Immunologie und Rheumatologie, Universitätsklinikum Tübingen, Tübingen, Germany. anne.wiesmann@med.uni-tuebingen.de

ABSTRACT

Background: Mesenchymal stem cells (MSC) are multipotent cells which can differentiate along osteogenic, chondrogenic, and adipogenic lineages. The present study was designed to investigate the influence of mechanical force as a specific physiological stress on the differentiation of (MSC) to osteoblast-like cells.

Methods: Human MSC were cultured in osteoinductive medium with or without cyclic uniaxial mechanical stimulation (2000 mustrain, 200 cycles per day, 1 Hz). Cultured cells were analysed for expression of collagen type I, osteocalcin, osteonectin, and CD90. To evaluate the biomineral formation the content of bound calcium in the cultures was determined.

Results: After 14 days in culture immunfluorescence staining revealed enhancement of collagen type I and osteonectin expression in response to mechanical stimulation. In contrast, mechanically stimulated cultures stained negative for CD90. In stimulated and unstimulated cultures an increase in the calcium content over time was observed. After 21 days in culture the calcium content in mechanical stimulated cultures was significantly higher compared to unstimulated control cultures.

Conclusion: These results demonstrate the influence of mechanical force on the differentiation of human MSC into osteoblast-like cells in vitro. While significant enhancement of the biomineral formation by mechanical stimulation is not detected before 21 days, effects on the extracellular matrix became already obvious after 14 days. The decrease of CD90 expression in mechanically stimulated cultures compared to unstimulated control cultures suggests that CD90 is only transiently expressed expression during the differentiation of MSC to osteoblast-like cells in culture.

No MeSH data available.


Related in: MedlinePlus