Calcified Tissue International

, Volume 101, Issue 3, pp 312–320 | Cite as

Potential of Osteoblastic Cells Derived from Bone Marrow and Adipose Tissue Associated with a Polymer/Ceramic Composite to Repair Bone Tissue

  • Gileade P. Freitas
  • Helena B. Lopes
  • Adriana L. G. Almeida
  • Rodrigo P. F. Abuna
  • Rossano Gimenes
  • Lucas E. B. Souza
  • Dimas T. Covas
  • Marcio M. Beloti
  • Adalberto L. RosaEmail author
Original Research


One of the tissue engineering strategies to promote bone regeneration is the association of cells and biomaterials. In this context, the aim of this study was to evaluate if cell source, either from bone marrow or adipose tissue, affects bone repair induced by osteoblastic cells associated with a membrane of poly(vinylidene-trifluoroethylene)/barium titanate (PVDF-TrFE/BT). Mesenchymal stem cells (MSC) were isolated from rat bone marrow and adipose tissue and characterized by detection of several surface markers. Also, both cell populations were cultured under osteogenic conditions and it was observed that MSC from bone marrow were more osteogenic than MSC from adipose tissue. The bone repair was evaluated in rat calvarial defects implanted with PVDF-TrFE/BT membrane and locally injected with (1) osteoblastic cells differentiated from MSC from bone marrow, (2) osteoblastic cells differentiated from MSC from adipose tissue or (3) phosphate-buffered saline. Luciferase-expressing osteoblastic cells derived from bone marrow and adipose tissue were detected in bone defects after cell injection during 25 days without difference in luciferin signal between cells from both sources. Corroborating the in vitro findings, osteoblastic cells from bone marrow combined with the PVDF-TrFE/BT membrane increased the bone formation, whereas osteoblastic cells from adipose tissue did not enhance the bone repair induced by the membrane itself. Based on these findings, it is possible to conclude that, by combining a membrane with cells in this rat model, cell source matters and that bone marrow could be a more suitable source of cells for therapies to engineer bone.


Animal model Bone Mesenchymal stem cells Bone marrow Adipose tissue 



Fabiana Rosseto de Moraes, Fabíola Singaretti de Oliveira, Milla Sprone Tavares, Roger Rodrigo Fernandes and Sebastião Carlos Bianco are acknowledged for their technical assistance during the experiments.


This research was supported by the National Council of Technological and Scientific Development (Grant # 456871/2013-6, CNPq - Brazil).

Compliance with Ethical Standards

Conflict of interest

Gileade P. Freitas, Helena B. Lopes, Adriana L. G. Almeida, Rodrigo P. F. Abuna, Rossano Gimenes, Lucas E. B. Souza, Dimas T. Covas, Marcio M. Beloti and Adalberto L. Rosa declare that they have no conflict of interest associated with this study and there has been no financial support that could have influenced our outcomes.

Human and Animal Rights and Informed Consent

All applicable international, national and/or institutional guidelines for the care and use of animals were followed. The Committee of Ethics in Animal Research of the School of Dentistry of Ribeirão Preto, University of São Paulo approved all animal procedures performed during the experiments.

Supplementary material

223_2017_282_MOESM1_ESM.tif (3.2 mb)
Supplementary material 1 (TIFF 3238 kb) FIG. S1. Three-dimensional reconstructed micro-CT image (A) and light microscopy (B) of an untreated rat calvarial bone defect at 4 weeks. No significant bone formation was noticed in the defect (A), which was filled with connective tissue (B). Alizarin red and Stevenel’s blue stain. Scale bar: A = 1.5 mm and B = 200 μm


  1. 1.
    Berner A, Reichert JC, Müller MB, Zellner J, Pfeifer C, Dienstknecht T, Nerlich M, Sommerville S, Dickinson IC, Schütz MA, Füchtmeier B (2012) Treatment of long bone defects and non-unions: from research to clinical practice. Cell Tissue Res 347:501–519CrossRefPubMedGoogle Scholar
  2. 2.
    Maruyama T, Jeong J, Sheu TJ, Hsu W (2016) Stem cells of the suture mesenchyme in craniofacial bone development, repair and regeneration. Nat Commun 7:10526CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Eap S, Keller L, Schiavi J, Huck O, Jacomine L, Fioretti F, Gauthier C, Sebastian V, Schwinté P, Benkirane-Jessel N (2015) A living thick nanofibrous implant bifunctionalized with active growth factor and stem cells for bone regeneration. Int J Nanomedicine 10:1061–1075PubMedPubMedCentralGoogle Scholar
  4. 4.
    Sicchieri LG, Crippa GE, de Oliveira PT, Beloti MM, Rosa AL (2012) Pore size regulates cell and tissue interactions with PLGA-CaP scaffolds used for bone engineering. J Tissue Eng Regen Med 6:155–162CrossRefPubMedGoogle Scholar
  5. 5.
    Pallesen L, Schou S, Aaboe M, Hjørting-Hansen E, Nattestad A, Melsen F (2002) Influence of particle size of autogenous bone grafts on the early stages of bone regeneration: a histologic and stereologic study in rabbit calvarium. Int J Oral Maxillofac Implants 17:498–506PubMedGoogle Scholar
  6. 6.
    Gimenes R, Zaghete MA, Bertolini M, Varela JA, Coelho LO, Silva NF Jr (2004) Composites PVDF-TrFE/BT used as bioactive membranes for enhancing bone regeneration. In: Bar-CohenY, editor. Proceedings of SPIE, Smart Structures and Materials, Vol. 5385. SPIE, Bellinghan, WA, pp 539–547Google Scholar
  7. 7.
    Lopes HB, Santos TD, de Oliveira FS, Freitas GP, de Almeida AL, Gimenes R, Rosa AL, Beloti MM (2014) Poly(vinylidene-trifluoroethylene)/barium titanate composite for in vivo support of bone formation. J Biomater Appl 29:104–112CrossRefPubMedGoogle Scholar
  8. 8.
    Teixeira LN, Crippa GE, Gimenes R, Zaghete MA, de Oliveira PT, Rosa AL, Beloti MM (2011) Response of human alveolar bone-derived cells to a novel poly(vinylidene fluoride-trifluoroethylene)/barium titanate membrane. J Mater Sci Mater Med 22:151–158CrossRefPubMedGoogle Scholar
  9. 9.
    Beloti MM, de Oliveira PT, Gimenes R, Zaghete MA, Bertolini MJ, Rosa AL (2006) In vitro biocompatibility of a novel membrane of the composite poly(vinylidene-trifluoroethylene)/barium titanate. J Biomed Mater Res A 79:282–288CrossRefPubMedGoogle Scholar
  10. 10.
    Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Friedenstein AJ, Petrakova KV, Kurolesova AI, Frolova GP (1968) Heterotopic of bone marrow. Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation 6:230–247CrossRefPubMedGoogle Scholar
  12. 12.
    Abuna RP, de Oliveira FS, Santos TS, Guerra TR, Rosa AL, Beloti MM (2016) Participation of TNF-α in inhibitory effects of adipocytes on osteoblast differentiation. J Cell Physiol 231:204–214CrossRefPubMedGoogle Scholar
  13. 13.
    Zhu Y, Liu T, Song K, Fan X, Ma X, Cui Z (2008) Adipose-derived stem cell: a better stem cell than BMSC. Cell Biochem Funct 26:664–675CrossRefPubMedGoogle Scholar
  14. 14.
    Rebelatto CK, Aguiar AM, Moretão MP, Senegaglia AC, Hansen P, Barchiki F, Oliveira J, Martins J, Kuligovski C, Mansur F, Christofis A, Amaral VF, Brofman PS, Goldenberg S, Nakao LS, Correa A (2008) Dissimilar differentiation of mesenchymal stem cells from bone marrow, umbilical cord blood, and adipose tissue. Exp Biol Med (Maywood) 233:901–913CrossRefGoogle Scholar
  15. 15.
    Dosier CR, Uhrig BA, Willett NJ, Krishnan L, Li MT, Stevens HY, Schwartz Z, Boyan BD, Guldberg RE (2015) Effect of cell origin and timing of delivery for stem cell-based bone tissue engineering using biologically functionalized hydrogels. Tissue Eng Part A 21:156–165CrossRefPubMedGoogle Scholar
  16. 16.
    Beloti MM, Sicchieri LG, de Oliveira PT, Rosa AL (2012) The influence of osteoblast differentiation stage on bone formation in autogenously implanted cell-based poly(lactide-co-glycolide) and calcium phosphate constructs. Tissue Eng Part A 18:999–1005CrossRefPubMedGoogle Scholar
  17. 17.
    Barzilay R, Sadan O, Melamed E, Offen D (2009) Comparative characterization of bone marrow-derived mesenchymal stromal cells from four different rat strains. Cytotherapy 11:435–442CrossRefPubMedGoogle Scholar
  18. 18.
    Harting M, Jimenez F, Pati S, Baumgartner J, Cox C Jr (2008) Immunophenotype characterization of rat mesenchymal stromal cells. Cytotherapy 10:243–253CrossRefPubMedGoogle Scholar
  19. 19.
    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop DJ, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317CrossRefPubMedGoogle Scholar
  20. 20.
    Han J, Koh YJ, Moon HR, Ryoo HG, Cho CH, Kim I, Koh GY (2010) Adipose tissue is an extramedullary reservoir for functional hematopoietic stem and progenitor cells. Blood 115:957–964CrossRefPubMedGoogle Scholar
  21. 21.
    Pachón-Peña G, Yu G, Tucker A, Wu X, Vendrell J, Bunnell BA, Gimble JM (2011) Stromal stem cells from adipose tissue and bone marrow of age-matched female donors display distinct immunophenotypic profiles. J Cell Physiol 226:843–851CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Shafiee A, Seyedjafari E, Soleimani M, Ahmadbeigi N, Dinarvand P, Ghaemi N (2011) A comparison between osteogenic differentiation of human unrestricted somatic stem cells and mesenchymal stem cells from bone marrow and adipose tissue. Biotechnol Lett 33:1257–1264CrossRefPubMedGoogle Scholar
  23. 23.
    Post S, Abdallah BM, Bentzon JF, Kassem M (2008) Demonstration of the presence of independent pre-osteoblastic and pre-adipocytic cell populations in bone marrow-derived mesenchymal stem cells. Bone 43:32–39CrossRefPubMedGoogle Scholar
  24. 24.
    Sakaguchi Y, Sekiya I, Yagishita K, Muneta T (2005) Comparison of human stem cells derived from various mesenchymal tissues: superiority of synovium as a cell source. Arthritis Rheum 52:2521–2529CrossRefPubMedGoogle Scholar
  25. 25.
    Kyllönen L, Haimi S, Mannerström B, Huhtala H, Rajala KM, Skottman H, Sándor GK, Miettinen S (2013) Effects of different serum conditions on osteogenic differentiation of human adipose stem cells in vitro. Stem Cell Res Ther 4:17CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Liao Y, Zhang XL, Li L, Shen FM, Zhong MK (2014) Stem cell therapy for bone repair: a systematic review and meta-analysis of preclinical studies with large animal models. Br J Clin Pharmacol 78:718–726CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Lebouvier A, Poignard A, Cavet M, Amiaud J, Leotot J, Hernigou P, Rahmouni A, Bierling P, Layrolle P, Rouard H, Chevallier N (2015) Development of a simple procedure for the treatment of femoral head osteonecrosis with intra-osseous injection of bone marrow mesenchymal stromal cells: study of their biodistribution in the early time points after injection. Stem Cell Res Ther 6:68CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Detante O, Moisan A, Dimastromatteo J, Richard MJ, Riou L, Grillon E, Barbier E, Desruet MD, De Fraipont F, Segebarth C, Jaillard A, Hommel M, Ghezzi C, Remy C (2009) Intravenous administration of 99mTc-HMPAO-labeled human mesenchymal stem cells after stroke: in vivo imaging and biodistribution. Cell Transplant 18:1369–1379CrossRefPubMedGoogle Scholar
  29. 29.
    Han DS, Chang HK, Kim KR, Woo SM (2014) Consideration of bone regeneration effect of stem cells: comparison of bone regeneration between bone marrow stem cells and adipose-derived stem cells. J Craniofac Surg 25:196–201CrossRefPubMedGoogle Scholar
  30. 30.
    Stockmann P, Park J, von Wilmowsky C, Nkenke E, Felszeghy E, Dehner JF, Schmitt C, Tudor C, Schlegel KA (2012) Guided bone regeneration in pig calvarial bone defects using autologous mesenchymal stem/progenitor cells—a comparison of different tissue sources. J Craniomaxillofac Surg 40:310–320CrossRefPubMedGoogle Scholar
  31. 31.
    Zhang W, Zhang X, Wang S, Xu L, Zhang M, Wang G, Jin Y, Zhang X, Jiang X (2013) Comparison of the use of adipose tissue-derived and bone marrow-derived stem cells for rapid bone regeneration. J Dent Res 92:1136–1141CrossRefPubMedGoogle Scholar
  32. 32.
    Hayashi O, Katsube Y, Hirose M, Ohgushi H, Ito H (2008) Comparison of osteogenic ability of rat mesenchymal stem cells from bone marrow, periosteum, and adipose tissue. Calcif Tissue Int 82:238–247CrossRefPubMedGoogle Scholar
  33. 33.
    Liu TM, Martina M, Hutmacher DW, Hui JH, Lee EH, Lim B (2007) Identification of common pathways mediating differentiation of bone marrow- and adipose tissue-derived human mesenchymal stem cells into three mesenchymal lineages. Stem Cells 25:750–760CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Gileade P. Freitas
    • 1
  • Helena B. Lopes
    • 1
  • Adriana L. G. Almeida
    • 1
  • Rodrigo P. F. Abuna
    • 1
  • Rossano Gimenes
    • 2
  • Lucas E. B. Souza
    • 3
    • 4
  • Dimas T. Covas
    • 3
    • 4
  • Marcio M. Beloti
    • 1
  • Adalberto L. Rosa
    • 1
    Email author
  1. 1.Cell Culture Laboratory, School of Dentistry of Ribeirão PretoUniversity of São PauloRibeirão PretoBrazil
  2. 2.Institute of Physics and ChemistryFederal University of ItajubáItajubáBrazil
  3. 3.National Institute of Science and Technology in Stem Cell and Cell TherapyRibeirão PretoBrazil
  4. 4.Department of Clinical Medicine, Ribeirão Preto School of MedicineUniversity of São PauloRibeirão PretoBrazil

Personalised recommendations