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International Orthopaedics

, Volume 43, Issue 1, pp 25–34 | Cite as

Bone marrow concentrate and expanded mesenchymal stromal cell surnatants as cell-free approaches for the treatment of osteochondral defects in a preclinical animal model

  • Francesca Veronesi
  • Giovanna Desando
  • Milena Fini
  • Annapaola Parrilli
  • Roberta Lolli
  • Melania Maglio
  • Lucia Martini
  • Gianluca Giavaresi
  • Isabella Bartolotti
  • Brunella Grigolo
  • Maria Sartori
Original Paper
  • 131 Downloads

Abstract

Purpose

To evaluate the regenerative potential of surnatants (SNs) from bone marrow concentrate (SN-BMC) and expanded mesenchymal stromal cells (SN-MSCs) loaded onto a collagen scaffold (SC) in comparison with cell-based treatments (BMC and MSCs) in an osteochondral (OC) defect model in rabbits.

Methods

OC defects (3 × 5 mm) were created in the rabbit femoral condyles and treated with SC alone or combined with SN-BMC, SN-MSCs, BMC, and MSCs. In control groups, the defects were left untreated. At three and six months, the quality of regenerated tissue was evaluated with macroscopic, histologic, microtomographic, and immunohistochemical assessments. The production of several immunoenzymatic markers was measured in the synovial fluid.

Results

All proposed treatments improved OC regeneration in comparison with untreated and SC-treated defects. Both BMC and MSCs showed a similar healing potential than their respective SNs, with the best performance exerted by BMC as demonstrated with macroscopic and histological scores and type I and II collagen results.

Conclusions

SNs loaded onto SC exerted a positive effect on OC defect regeneration, underlying the biological significance of the trophic factors, thus potentially opening new opportunities for the use of cell-free-based therapies. BMC was confirmed to be the most beneficial treatment.

Keywords

Osteochondral defect In vivo model Cell-free approach Bone marrow concentrate Mesenchymal stromal cells 

Notes

Funding information

This work was partially supported by the Ministry of Health-Ricerca Corrente to the IRCCS Rizzoli Orthopaedic Institute and by a grant from Regione Emilia Romagna: Programma di Ricerca Regione-Università 2010–2012—Strategic Program “Regenerative Medicine of Cartilage and Bone” (PRUa1RI-2012-007).

Compliance with ethical standards

The experimental protocol and surgical procedures were approved by a local Ethical Committee and authorized by the Italian Ministry of Health (Title of the project: One-step surgery with stem cells for the treatment of osteochondral lesions—No. 0017661, approved on May 29, 2013).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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References

  1. 1.
    Hofer HR, Tuan RS (2016) Secreted trophic factors of mesenchymal stem cells support neurovascular and musculoskeletal therapies. Stem Cell Res Ther 7(131).  https://doi.org/10.1186/s13287-016-0394-0
  2. 2.
    Cavallo C, Desando G, Ferrari A, Zini N, Mariani E, Grigolo B (2016) Hyaluronan scaffold supports osteogenic differentiation of bone marrow concentrate cells. J Biol Regul Homeost Agents 30:409–420Google Scholar
  3. 3.
    Veronesi F, Giavaresi G, Tschon M, Borsari V, Nicoli Aldini N, Fini M (2013) Clinical use of bone marrow, bone marrow concentrate, and expanded bone marrow mesenchymal stem cells in cartilage disease. Stem Cells Dev 22:181–192.  https://doi.org/10.1089/scd.2012.0373 CrossRefGoogle Scholar
  4. 4.
    Sartori M, Pagani S, Ferrari A, Costa V, Carina V, Figallo E, Maltarello MC, Martini L, Fini M, Giavaresi G (2017) A new bi-layered scaffold for osteochondral tissue regeneration: in vitro and in vivo preclinical investigations. Mater Sci Eng C Mater Biol Appl 70:101–111.  https://doi.org/10.1016/j.msec.2016.08.027 CrossRefGoogle Scholar
  5. 5.
    Veronesi F, Torricelli P, Borsari V, Tschon M, Rimondini L, Fini M (2011) Mesenchymal stem cells in the aging and osteoporotic population. Crit Rev Eukaryot Gene Expr 21:363–377CrossRefGoogle Scholar
  6. 6.
    Desando G, Giavaresi G, Cavallo C, Bartolotti I, Sartoni F, Nicoli Aldini N, Martini L, Parrilli A, Mariani E, Fini M, Grigolo B (2016) Autologous bone marrow concentrate in a sheep model of osteoarthritis: new perspectives for cartilage and meniscus repair. Tissue Eng Part C Methods 22:608–619.  https://doi.org/10.1089/ten.TEC.2016.0033 CrossRefGoogle Scholar
  7. 7.
    Veronesi F, Cadossi M, Giavaresi G, Martini L, Setti S, Buda R, Giannini S, Fini M (2015) Pulsed electromagnetic fields combined with a collagenous scaffold and bone marrow concentrate enhance osteochondral regeneration: an in vivo study. BMC Musculoskelet Disord 16(233).  https://doi.org/10.1186/s12891-015-0683-2
  8. 8.
    Desando G, Bartolotti I, Vannini F, Cavallo C, Castagnini F, Buda R, Giannini S, Mosca M, Mariani E, Grigolo B (2017) Repair potential of matrix-induced bone marrow aspirate concentrate and matrix-induced autologous chondrocyte implantation for talar osteochondral repair: patterns of some catabolic, inflammatory, and pain mediators. Cartilage 8:50–60CrossRefGoogle Scholar
  9. 9.
    Giannini S, Buda R, Vannini F, Cavallo M, Grigolo B (2009) One-step bone marrow-derived cell transplantation in talar osteochondral lesions. Clin Orthop Relat Res 467:3307–3320.  https://doi.org/10.1007/s11999-009-0885-8 CrossRefGoogle Scholar
  10. 10.
    Caplan AI, Dennis JE (2006) Mesenchymal stem cells as trophic mediators. J Cell Biochem 98:1076–1084CrossRefGoogle Scholar
  11. 11.
    Veronesi F, Borsari V, Sartori M, Orciani M, Mattioli-Belmonte M, Fini M (2018) The use of cell conditioned medium for musculoskeletal tissue regeneration. J Cell Physiol 233:4423–4442.  https://doi.org/10.1002/jcp.26291 CrossRefGoogle Scholar
  12. 12.
    Cavallo C, Desando G, Cattini L, Cavallo M, Buda R, Giannini S, Facchini A, Grigolo B (2013) Bone marrow concentrated cell transplantation: rationale for its use in the treatment of human osteochondral lesions. J Biol Regul Homeost Agents 27:165–175Google Scholar
  13. 13.
    Brittberg M, Winalski CS (2003) Evaluation of cartilage injuries and repair. J Bone Joint Surg Am 85-A(Suppl 2):58–69CrossRefGoogle Scholar
  14. 14.
    O’Driscoll SW, Keeley FW, Salter RB (1986) The chondrogenic potential of free autogenous periosteal grafts for biological resurfacing of major full-thickness defects in joint surfaces under the influence of continuous passive motion. An experimental investigation in the bone. J Bone Joint Surg Am 68:1017–1035CrossRefGoogle Scholar
  15. 15.
    R Development Core Team (2008) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0, URL http://www.R-project.org
  16. 16.
    Im GI (2017) Clinical use of stem cells in orthopaedics. Eur Cell Mater 33:183–196.  https://doi.org/10.22203/eCM.v033a14 CrossRefGoogle Scholar
  17. 17.
    Welch T, Mandelbaum B, Tom M (2016) Autologous chondrocyte implantation: past, present, and future. Sports Med Arthrosc Rev 24:85–91.  https://doi.org/10.1097/JSA.0000000000000115 CrossRefGoogle Scholar
  18. 18.
    Hochrein A, Zinser W, Spahn G, Angele P, Löer I, Albrecht D, Niemeyer P (2018) What parameters affect knee function in patients with untreated cartilage defects: baseline data from the German Cartilage Registry. Int Orthop.  https://doi.org/10.1007/s00264-018-4125-2
  19. 19.
    Volz M, Schaumburger J, Frick H, Grifka J, Anders S (2017) A randomized controlled trial demonstrating sustained benefit of autologous matrix-induced Chondrogenesis over microfracture at five years. Int Orthop 41(4):797–804.  https://doi.org/10.1007/s00264-016-3391-0 CrossRefGoogle Scholar
  20. 20.
    Lavoie JR, Rosu-Myles (2013) Uncovering the secretes of mesenchymal stem cells. Biochimie 95:2212–2221.  https://doi.org/10.1016/j.biochi.2013.06.017 CrossRefGoogle Scholar
  21. 21.
    Sampson S, Botto-van Bemden A, Aufiero D (2013) Autologous bone marrow concentrate: review and application of a novel intra-articular orthobiologic for cartilage disease. Phys Sportsmed 41(3):7–18.  https://doi.org/10.3810/psm.2013.09.2022 CrossRefGoogle Scholar
  22. 22.
    Cicione C, Muinos-Lopez E, Hermida-Gomez T, Fuentes-Boquete I, Diaz-Prado S, Blanco FJ (2016) Alternative protocols to induce chondrogenic differentiation: transforming growth factor-beta superfamily. Cell Tissue Bank 16:195CrossRefGoogle Scholar
  23. 23.
    Hernigou J, Vertongen P, Chahidi E, Kyriakidis T, Dehoux JP, Crutzen M, Boutry S, Larbanoix L, Houben S, Gaspard N, Koulalis D, Rasschaert J (2018) Effects of press-fit biphasic (collagen and HA/βTCP) scaffold with cell-based therapy on cartilage and subchondral bone repair knee defect in rabbits. Int Orthop 42:1755–1767.  https://doi.org/10.1007/s00264-018-3999-3 CrossRefGoogle Scholar
  24. 24.
    Santo VE, Gomes ME, Mano JF, Reis RL (2013) Controlled release strategies for bone, cartilage, and osteochondral engineering--part II: challenges on the evolution from single to multiple bioactive factor delivery. Tissue Eng Part B Rev 19:327–352.  https://doi.org/10.1089/ten.TEB.2012.0138 CrossRefGoogle Scholar
  25. 25.
    Wang Q, Zhang H, Gan H, Wang H, Li Q, Wang Z (2018) Application of combined porous tantalum scaffolds loaded with bone morphogenetic protein 7 to repair of osteochondral defect in rabbits. Int Orthop 42(7):1437–1448.  https://doi.org/10.1007/s00264-018-3800-7 CrossRefGoogle Scholar
  26. 26.
    Zhang Z, Li L, Yang W, Cao Y, Shi Y, Li X, Zhang Q (2017) The effects of different doses of IGF-1 on cartilage and subchondral bone during the repair of full-thickness articular cartilage defects in rabbits. Osteoarthr Cartil 25:309–320.  https://doi.org/10.1016/j.joca.2016.09.010 CrossRefGoogle Scholar
  27. 27.
    Lin H, Hay E, Latourte A, Funck-Brentano T, Bouaziz W, Ea HK, Khatib AM, Richette P, Cohen-Solal M (2018) Proprotein convertase furin inhibits matrix metalloproteinase 13 in a TGFβ-dependent manner and limits osteoarthritis in mice. Sci Rep 8(10488).  https://doi.org/10.1038/s41598-018-28713-2
  28. 28.
    Chen B, Li Q, Zhao B, Wang Y (2017) Stem cell-derived extracellular vescicles as a novel potential therapeutic tool for tissue repair. Stem Cells Transl Med 6:1753–1758.  https://doi.org/10.1002/sctm.16-0477 CrossRefGoogle Scholar

Copyright information

© SICOT aisbl 2018

Authors and Affiliations

  • Francesca Veronesi
    • 1
  • Giovanna Desando
    • 2
  • Milena Fini
    • 1
  • Annapaola Parrilli
    • 1
  • Roberta Lolli
    • 1
  • Melania Maglio
    • 1
  • Lucia Martini
    • 1
  • Gianluca Giavaresi
    • 1
  • Isabella Bartolotti
    • 2
  • Brunella Grigolo
    • 2
  • Maria Sartori
    • 1
  1. 1.Laboratory of Preclinical and Surgical Studies, Rizzoli-RITIRCCS Rizzoli Orthopedic InstituteBolognaItaly
  2. 2.Laboratory RAMSESIRCCS Rizzoli Orthopedic InstituteBolognaItaly

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