Analysis of ectopic and orthopic bone formation in cell-based tissue-engineered constructs in goats

M.C. Kruyt, W.J.A. Dhert, F. Cumhur Oner, Clemens van Blitterswijk, Abraham J. Verbout, J.D. de Bruijn

    Research output: Contribution to journalArticleAcademicpeer-review

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    Abstract

    Despite decades of extensive research, the application of cell-based bone tissue engineering in clinically relevant models remains challenging. To improve effectiveness, a better understanding of how the technique should work is crucial. In the current study, we investigated the onset time, rate, location and direction of bone formation in ectopically and orthotopically implanted clinically sized tissue-engineered constructs to gain insight the mechanism behind it. Bone marrow stromal cells (BMSCs) were obtained from 10 goats, culture expanded and cryopreserved. Porous biphasic calcium phosphate (BCP) disks of 17 mm×6 mm were per-operatively seeded with BMSCs or left empty. Both conditions were implanted intramuscularly and in bilateral critical-sized iliac wing defects. Fluorochromes were administered at 3, 5 and 7 weeks and samples were retrieved after 9 weeks. Histology showed abundant and homogeneous bone formation throughout the intramuscular BMSC samples and little bone in the controls. Histomorphometry and measurements of the fluorochrome labels of the ectopical BMSC samples indicated that osteogenesis started at the periphery and subsequent osteoconduction filled the whole scaffold within 7 weeks. In the orthotopically implanted disks, there was good integration with the surrounding bone, but minimal bone in the center of the implants, in both conditions. Bone was only derived from the interface with the surrounding bone, there was no early bone at the surfaces in contact to soft tissue as was seen in the ectopical samples. Apparently cell survival was minimal and insufficient for relevant additional bone formation. However, the speed of integration with surrounding bone and subsequent bone apposition on the BMSC-seeded orthotopic scaffolds were found to be significantly enhanced, which may be relevant especially in challenging environments.
    Original languageEnglish
    Pages (from-to)1798-1805
    JournalBiomaterials
    Volume28
    Issue number10
    Early online date19 Dec 2006
    DOIs
    Publication statusPublished - Apr 2007

    Fingerprint

    Osteogenesis
    Goats
    Bone
    Mesenchymal Stromal Cells
    Tissue
    Bone and Bones
    Fluorescent Dyes
    Bone Regeneration
    Tissue Engineering
    Cell Survival
    Histology
    Scaffolds
    Research
    Calcium phosphate
    Tissue engineering
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    Keywords

    • Polychrome labeling
    • Cell survival
    • Stem cells
    • Goats
    • IR-78865
    • METIS-245445
    • Bone tissue engineering

    Cite this

    Kruyt, M. C., Dhert, W. J. A., Cumhur Oner, F., van Blitterswijk, C., Verbout, A. J., & de Bruijn, J. D. (2007). Analysis of ectopic and orthopic bone formation in cell-based tissue-engineered constructs in goats. Biomaterials, 28(10), 1798-1805. https://doi.org/10.1016/j.biomaterials.2006.11.038
    Kruyt, M.C. ; Dhert, W.J.A. ; Cumhur Oner, F. ; van Blitterswijk, Clemens ; Verbout, Abraham J. ; de Bruijn, J.D. . / Analysis of ectopic and orthopic bone formation in cell-based tissue-engineered constructs in goats. In: Biomaterials. 2007 ; Vol. 28, No. 10. pp. 1798-1805.
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    abstract = "Despite decades of extensive research, the application of cell-based bone tissue engineering in clinically relevant models remains challenging. To improve effectiveness, a better understanding of how the technique should work is crucial. In the current study, we investigated the onset time, rate, location and direction of bone formation in ectopically and orthotopically implanted clinically sized tissue-engineered constructs to gain insight the mechanism behind it. Bone marrow stromal cells (BMSCs) were obtained from 10 goats, culture expanded and cryopreserved. Porous biphasic calcium phosphate (BCP) disks of 17 mm×6 mm were per-operatively seeded with BMSCs or left empty. Both conditions were implanted intramuscularly and in bilateral critical-sized iliac wing defects. Fluorochromes were administered at 3, 5 and 7 weeks and samples were retrieved after 9 weeks. Histology showed abundant and homogeneous bone formation throughout the intramuscular BMSC samples and little bone in the controls. Histomorphometry and measurements of the fluorochrome labels of the ectopical BMSC samples indicated that osteogenesis started at the periphery and subsequent osteoconduction filled the whole scaffold within 7 weeks. In the orthotopically implanted disks, there was good integration with the surrounding bone, but minimal bone in the center of the implants, in both conditions. Bone was only derived from the interface with the surrounding bone, there was no early bone at the surfaces in contact to soft tissue as was seen in the ectopical samples. Apparently cell survival was minimal and insufficient for relevant additional bone formation. However, the speed of integration with surrounding bone and subsequent bone apposition on the BMSC-seeded orthotopic scaffolds were found to be significantly enhanced, which may be relevant especially in challenging environments.",
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    Kruyt, MC, Dhert, WJA, Cumhur Oner, F, van Blitterswijk, C, Verbout, AJ & de Bruijn, JD 2007, 'Analysis of ectopic and orthopic bone formation in cell-based tissue-engineered constructs in goats' Biomaterials, vol. 28, no. 10, pp. 1798-1805. https://doi.org/10.1016/j.biomaterials.2006.11.038

    Analysis of ectopic and orthopic bone formation in cell-based tissue-engineered constructs in goats. / Kruyt, M.C.; Dhert, W.J.A.; Cumhur Oner, F.; van Blitterswijk, Clemens; Verbout, Abraham J.; de Bruijn, J.D. .

    In: Biomaterials, Vol. 28, No. 10, 04.2007, p. 1798-1805.

    Research output: Contribution to journalArticleAcademicpeer-review

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    KW - Polychrome labeling

    KW - Cell survival

    KW - Stem cells

    KW - Goats

    KW - IR-78865

    KW - METIS-245445

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    Kruyt MC, Dhert WJA, Cumhur Oner F, van Blitterswijk C, Verbout AJ, de Bruijn JD. Analysis of ectopic and orthopic bone formation in cell-based tissue-engineered constructs in goats. Biomaterials. 2007 Apr;28(10):1798-1805. https://doi.org/10.1016/j.biomaterials.2006.11.038