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In Vivo Bone Formation Within Engineered Hydroxyapatite Scaffolds in a Sheep Model

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Abstract

Large bone defects still represent a major burden in orthopedics, requiring bone-graft implantation to promote the bone repair. Along with autografts that currently represent the gold standard for complicated fracture repair, the bone tissue engineering offers a promising alternative strategy combining bone-graft substitutes with osteoprogenitor cells able to support the bone tissue ingrowth within the implant. Hence, the optimization of cell loading and distribution within osteoconductive scaffolds is mandatory to support a successful bone formation within the scaffold pores. With this purpose, we engineered constructs by seeding and culturing autologous, osteodifferentiated bone marrow mesenchymal stem cells within hydroxyapatite (HA)-based grafts by means of a perfusion bioreactor to enhance the in vivo implant-bone osseointegration in an ovine model. Specifically, we compared the engineered constructs in two different anatomical bone sites, tibia, and femur, compared with cell-free or static cell-loaded scaffolds. After 2 and 4 months, the bone formation and the scaffold osseointegration were assessed by micro-CT and histological analyses. The results demonstrated the capability of the acellular HA-based grafts to determine an implant-bone osseointegration similar to that of statically or dynamically cultured grafts. Our study demonstrated that the tibia is characterized by a lower bone repair capability compared to femur, in which the contribution of transplanted cells is not crucial to enhance the bone-implant osseointegration. Indeed, only in tibia, the dynamic cell-loaded implants performed slightly better than the cell-free or static cell-loaded grafts, indicating that this is a valid approach to sustain the bone deposition and osseointegration in disadvantaged anatomical sites.

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Acknowledgments

This work was funded by Regione Lombardia with POR FESR 2007-2013 resources (Grant ID: ATP 2009, No. 13396272). The authors thank Finceramica for providing the Engipore® scaffolds.

Author contribution

Guarantor: author#9. Study concept and design: authors #1, #2, #3, #4, #8 and #9. Data acquisition and analyses: authors #1, #2, #3, #4, #5 and #7. Data interpretation: authors #1, #2, #3, #4 and #6. Drafting of the manuscript: authors #1, #2, #3, #4, #5, #6 and #7. Critical revision of the manuscript for important intellectual contents: authors #1, #2, #3, #6, #8 and #9. All authors approved the final version of the manuscript.

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    Authors and Affiliations

    1. Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Milan, Italy

      A. B. Lovati, S. Lopa, G. Talò, M. Bottagisio & M. Moretti

    2. Mouse and Animal Pathology Laboratory, Fondazione Filarete, Milan, Italy

      C. Recordati, M. Losa & E. Scanziani

    3. Dipartimento di Elettronica, Informazione e Bioingegneria (DEIB), Politecnico di Milano, Milan, Italy

      C. Turrisi

    4. Department of Veterinary Sciences and Public Health, University of Milan, Milan, Italy

      M. Bottagisio & E. Scanziani

    5. Regenerative Medicine Technologies Lab, Ente Ospedaliero Cantonale (EOC), Lugano, Switzerland

      M. Moretti

    6. Swiss Institute of Regenerative Medicine (SIRM), Lugano, Switzerland

      M. Moretti

    7. Fondazione Cardiocentro Ticino, Lugano, Switzerland

      M. Moretti

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    1. A. B. Lovati
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    Correspondence to M. Moretti.

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    Conflict of interest

    M Moretti has received research grant from Regione Lombardia with POR FESR 2007–2013 resources (Grant ID: ATP 2009, No. 13396272) and he is inventor of the patent #WO2008098165A2 issued by USPTO.

    Human and Animal Rights and Informed Consent

    This article does not contain studies with human participants performed by any of the authors. All applicable international, national and/or institutional guidelines for the care and use of animals were followed. Specifically, the Lazzaro Spallanzani Institute Animal Care and Use Committee (IACUC) approved the whole study. Animals and their care were handled in compliance with institutional guidelines as defined in national (Law 116/92, Authorization n.19/2008-A issued March 6, 2008, by the Italian Ministry of Health) and international laws and policies (EEC Council Directive 86/609, OJ L 358. 1, December 12, 1987; Standards for the Care and Use of Laboratory Animals—UCLA, U. S. National Research Council, Statement of Compliance A5023-01, November 6, 1998). The animals were housed at the facilities of the Lazzaro Spallanzani Institute according international standards. A certified veterinarian—responsible for health monitoring, animal welfare supervision, experimental protocols and procedure revision—regularly checked animals. All procedures on animals performed in this study were in accordance with the ethical standards.

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    S. Lopa and C. Recordati have contributed equally to this work.

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    Lovati, A.B., Lopa, S., Recordati, C. et al. In Vivo Bone Formation Within Engineered Hydroxyapatite Scaffolds in a Sheep Model. Calcif Tissue Int 99, 209–223 (2016). https://doi.org/10.1007/s00223-016-0140-8

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    • DOI: https://doi.org/10.1007/s00223-016-0140-8

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