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Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells

  • Authors:
    • Dirk Henrich
    • Christoph Nau
    • Saskia Bo Kraft
    • Maximilian Zollfrank
    • Kerstin Kontradowitz
    • Elsie Oppermann
    • Judith Schultheiss
    • Simon Meier
    • Johannes Frank
    • Ingo Marzi
    • Caroline Seebach
  • View Affiliations / Copyright

    Affiliations: Department of Trauma, Hand and Reconstructive Surgery, Goethe University Hospital, 60590 Frankfurt am Main, Germany, Department of Visceral and Abdominal Surgery, Goethe University Hospital, 60590 Frankfurt am Main, Germany
    Copyright: © Henrich et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 976-988
    |
    Published online on: February 12, 2016
       https://doi.org/10.3892/ijmm.2016.2489
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Abstract

Evidence has indicated that mesenchymal stem cells (MSCs) harvested with the Reamer/Irrigator/Aspirator (RIA) procedure exhibited an improved osteogenic differentiation capability compared with MSCs obtained by bone marrow aspiration from the iliac crest. In the present study, we hypothesized that the harvest procedure indeed influences the osteogenic activity of human MSCs more than the tissue site itself. Concentration [by colony forming unit-fibroblast (CFU-F) assay], calcification (by von Kossa staining), collagen deposition, gene expression and the gene methylation of the bone morphogenetic protein (BMP)-2 pathway [BMP2, SMAD5 and runt-related transcription factor 2 (RUNX2)], the Wnt pathway [WNT3, dickkopf-1 (DKK1), low-density lipoprotein receptor‑related protein 5 (LRP5) and β-catenin] and osteogenic genes [alkaline phosphatase (ALP), collagen, type I, alpha 1 (COL1A) and osteocalcin] were analyzed in the MSCs isolated intraoperatively from the iliac crest with a spoon (n=14), from the femur with a spoon (n=7), from the femur with the RIA procedure (n=13) and from the iliac crest by fine-needle aspiration (n=8, controls). A Bonferroni-Holm corrected p-value <0.05 indicated a statistically significant difference. The concentration of CFU-F in the MSCs was increased in the RIA debris in comparison with that in the iliac crest aspirates (trend) and the femur (spoon, significant). Calcium deposition was highest in the femur-derived MSCs (by RIA) and was significantly increased in comparison with that in the iliac crest-derived MSCs (spoon, aspirate). The gene expression of BMP2, SMAD5, RUNX2, osteocalcin, and COL1A was significantly increased in the femur-derived MSCs (spoon) and the iliac crest aspirate derived-MSCs in comparison with that in the femur-derived MSCs (by RIA). There was no significant diversity between the samples obtained using a spoon (from the femur or iliac crest). Calcium deposition and osteogenic gene expression decreased significantly with the increasing passage number in all the samples. The methylation of genes did not correlate with their respective gene expression and inconsistent differences were observed between the groups. Herein, we provide evidence that the harvest procedure is a critical factor in the osteogenesis of MSCs in vitro. The MSCs isolated from the femur and iliac crest using a spoon exhibit no significant differences. The altered gene expression and function of the femur-derived MSCs (by RIA) may be due to the harsh isolation procedure. The variable differentiation ability of the MSCs, which depends on the harvest site and the harvest technique, as well as the rapid loss of the osteogenic differentiation capacity with the increasing culture duration should be taken into consideration when using MSCs as a potential therapeutic application for bone tissue engineering.
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Copy and paste a formatted citation
Spandidos Publications style
Henrich D, Nau C, Kraft SB, Zollfrank M, Kontradowitz K, Oppermann E, Schultheiss J, Meier S, Frank J, Marzi I, Marzi I, et al: Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells. Int J Mol Med 37: 976-988, 2016.
APA
Henrich, D., Nau, C., Kraft, S.B., Zollfrank, M., Kontradowitz, K., Oppermann, E. ... Seebach, C. (2016). Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells. International Journal of Molecular Medicine, 37, 976-988. https://doi.org/10.3892/ijmm.2016.2489
MLA
Henrich, D., Nau, C., Kraft, S. B., Zollfrank, M., Kontradowitz, K., Oppermann, E., Schultheiss, J., Meier, S., Frank, J., Marzi, I., Seebach, C."Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells". International Journal of Molecular Medicine 37.4 (2016): 976-988.
Chicago
Henrich, D., Nau, C., Kraft, S. B., Zollfrank, M., Kontradowitz, K., Oppermann, E., Schultheiss, J., Meier, S., Frank, J., Marzi, I., Seebach, C."Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells". International Journal of Molecular Medicine 37, no. 4 (2016): 976-988. https://doi.org/10.3892/ijmm.2016.2489
Copy and paste a formatted citation
x
Spandidos Publications style
Henrich D, Nau C, Kraft SB, Zollfrank M, Kontradowitz K, Oppermann E, Schultheiss J, Meier S, Frank J, Marzi I, Marzi I, et al: Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells. Int J Mol Med 37: 976-988, 2016.
APA
Henrich, D., Nau, C., Kraft, S.B., Zollfrank, M., Kontradowitz, K., Oppermann, E. ... Seebach, C. (2016). Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells. International Journal of Molecular Medicine, 37, 976-988. https://doi.org/10.3892/ijmm.2016.2489
MLA
Henrich, D., Nau, C., Kraft, S. B., Zollfrank, M., Kontradowitz, K., Oppermann, E., Schultheiss, J., Meier, S., Frank, J., Marzi, I., Seebach, C."Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells". International Journal of Molecular Medicine 37.4 (2016): 976-988.
Chicago
Henrich, D., Nau, C., Kraft, S. B., Zollfrank, M., Kontradowitz, K., Oppermann, E., Schultheiss, J., Meier, S., Frank, J., Marzi, I., Seebach, C."Effect of the harvest procedure and tissue site on the osteogenic function of and gene expression in human mesenchymal stem cells". International Journal of Molecular Medicine 37, no. 4 (2016): 976-988. https://doi.org/10.3892/ijmm.2016.2489
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