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Regeneration of hyaline cartilage promoted by xenogeneic mesenchymal stromal cells embedded within elastin-like recombinamer-based bioactive hydrogels

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Abstract

Over the last decades, novel therapeutic tools for osteochondral regeneration have arisen from the combination of mesenchymal stromal cells (MSCs) and highly specialized smart biomaterials, such as hydrogel-forming elastin-like recombinamers (ELRs), which could serve as cell-carriers. Herein, we evaluate the delivery of xenogeneic human MSCs (hMSCs) within an injectable ELR-based hydrogel carrier for osteochondral regeneration in rabbits. First, a critical-size osteochondral defect was created in the femora of the animals and subsequently filled with the ELR-based hydrogel alone or with embedded hMSCs. Regeneration outcomes were evaluated after three months by gross assessment, magnetic resonance imaging and computed tomography, showing complete filling of the defect and the de novo formation of hyaline-like cartilage and subchondral bone in the hMSC-treated knees. Furthermore, histological sectioning and staining of every sample confirmed regeneration of the full cartilage thickness and early subchondral bone repair, which was more similar to the native cartilage in the case of the cell-loaded ELR-based hydrogel. Overall histological differences between the two groups were assessed semi-quantitatively using the Wakitani scale and found to be statistically significant (p < 0.05). Immunofluorescence against a human mitochondrial antibody three months post-implantation showed that the hMSCs were integrated into the de novo formed tissue, thus suggesting their ability to overcome the interspecies barrier. Hence, we conclude that the use of xenogeneic MSCs embedded in an ELR-based hydrogel leads to the successful regeneration of hyaline cartilage in osteochondral lesions.

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References

  1. 1.

    Kuettner KE, Cole AA. Cartilage degeneration in different human joints. Osteoarthr Cartil. 2005;13(2):93–103.

  2. 2.

    Hiligsmann M, Reginster J-Y. The economic weight of osteoarthritis in Europe. Medicographia. 2013;35(1):197–202.

  3. 3.

    Chevalier X. Physiopathology of arthrosis. The normal cartilage. Presse Med. 1998;27(2):75–80.

  4. 4.

    Vaquero J, Forriol F. Knee chondral injuries: clinical treatment strategies and experimental models. Injury. 2011;43:694–705.

  5. 5.

    Chen F, Rousche K, Tuan R. Technology insight: adult stem cells in cartilage regeneration and tissue engineering. Nat Clin Pract Rheumatol. 2006;2(7):373–82.

  6. 6.

    Li J, Ezzelarab MB, Cooper DKC. Do mesenchymal stem cells function across species barriers? Relevance for xenotransplantation. Xenotransplantation. 2012;19(5):273–85.

  7. 7.

    Roche ET, Hastings CL, Lewin SA, Shvartsman D, Brudno Y, Vasilyev NV, et al. Comparison of biomaterial delivery vehicles for improving acute retention of stem cells in the infarcted heart. Biomaterials. 2014;35(25):6850–8.

  8. 8.

    Martens TP, Godier AFG, Parks JJ, Wan LQ, Koeckert MS, Eng GM, et al. Percutaneous cell delivery into the heart using hydrogels polymerizing in situ. Cell Transplant. 2009;18(3):297–304.

  9. 9.

    Girotti A, Orbanic D, Ibáñez-Fonseca A, Gonzalez-Obeso C, Rodríguez-Cabello JC. Recombinant technology in the development of materials and systems for soft-tissue repair. Adv Healthc Mater. 2015;4(16):2423–55.

  10. 10.

    Rodriguez-Cabello JC, Martin L, Girotti A, Garcia-Arevalo C, Arias FJ, Alonso M. Emerging applications of multifunctional elastin-like recombinamers. Nanomedicine. 2011;6(1):111–22.

  11. 11.

    Urry DW. Molecular machines - how motion and other functions of living organisms can result from reversible chemical-changes. Angew Chem-Int Ed Engl. 1993;32(6):819–41.

  12. 12.

    Martin L, Arias FJ, Alonso M, Garcia-Arevalo C, Rodriguez-Cabello JC. Rapid micropatterning by temperature-triggered reversible gelation of a recombinant smart elastin-like tetrablock-copolymer. Soft Matter. 2010;6(6):1121–4.

  13. 13.

    Ruoslahti E, Pierschbacher MD. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986;44(4):517–8.

  14. 14.

    Trattnig S, Millington SA, Szomolanyi P, Marlovits S. MR imaging of osteochondral grafts and autologous chondrocyte implantation. Eur Radiol. 2007;17(1):103–18.

  15. 15.

    Barber FA, Dockery WD. A computed tomography scan assessment of synthetic multiphase polymer scaffolds used for osteochondral defect repair. Arthroscopy. 2011;27(1):60–4.

  16. 16.

    Orth P, Zurakowski D, Wincheringer D, Madry H. Reliability, reproducibility, and validation of five major histological scoring systems for experimental articular cartilage repair in the rabbit model. Tissue Eng Part C Methods. 2011;18(5):329–39.

  17. 17.

    Pérez-Simon JA, López-Villar O, Andreu EJ, Rifón J, Muntion S, Campelo MD, et al. Mesenchymal stem cells expanded in vitro with human serum for the treatment of acute and chronic graft-versus-host disease: results of a phase I/II clinical trial. Haematologica. 2011;96(7):1072–6.

  18. 18.

    Villaron E, Almeida J, Lopez-Holgado N, Alcoceba M, Sanchez-Abarca L, Sanchez-Guijo F, et al. Mesenchymal stem cells are present in peripheral blood and can engraft after allogeneic hematopoietic stem cell transplantation. Haematologica. 2004;89(12):1421–7.

  19. 19.

    Rodriguez-Cabello JC, Girotti A, Ribeiro A, Arias FJ. Synthesis of genetically engineered protein polymers (recombinamers) as an example of advanced self-assembled smart materials. Methods Mol Biol. 2012;811:17–38.

  20. 20.

    Nakamura T, Sekiya I, Muneta T, Hatsushika D, Horie M, Tsuji K, et al. Arthroscopic, histological and MRI analyses of cartilage repair after a minimally invasive method of transplantation of allogeneic synovial mesenchymal stromal cells into cartilage defects in pigs. Cytotherapy. 2012;14(3):327–38.

  21. 21.

    Hrabchak C, Rouleau J, Moss I, Woodhouse K, Akens M, Bellingham C, et al. Assessment of biocompatibility and initial evaluation of genipin cross-linked elastin-like polypeptides in the treatment of an osteochondral knee defect in rabbits. Acta Biomater. 2010;6(6):2108–15.

  22. 22.

    Nettles DL, Kitaoka K, Hanson NA, Flahiff CM, Mata BA, Hsu EW, et al. In situ crosslinking elastin-like polypeptide gels for application to articular cartilage repair in a goat osteochondral defect model. Tissue Eng Part A. 2008;14(7):1133–40.

  23. 23.

    Schütz K, Despang F, Lode A, Gelinsky M. Cell-laden biphasic scaffolds with anisotropic structure for the regeneration of osteochondral tissue. J Tissue Eng Regen Med. 2016;10(5):404–17.

  24. 24.

    Marquass B, Somerson JS, Hepp P, Aigner T, Schwan S, Bader A, et al. A novel MSC-seeded triphasic construct for the repair of osteochondral defects. J Orthop Res. 2010;28(12):1586–99.

  25. 25.

    Yang J, Zhang YS, Yue K, Khademhosseini A. Cell-laden hydrogels for osteochondral and cartilage tissue engineering. Acta Biomaterialia. 2017. doi:10.1016/j.actbio.2017.01.036.

  26. 26.

    Kim M, Foo LF, Uggen C, Lyman S, Ryaby JT, Moynihan DP, et al. Evaluation of early osteochondral defect repair in a rabbit model utilizing fourier transform-infrared imaging spectroscopy, magnetic resonance imaging, and quantitative T2 mapping. Tissue Eng Part C Methods. 2010;16(3):355–64.

  27. 27.

    Haleem AM, Singergy AA, Sabry D, Atta HM, Rashed LA, Chu CR, et al. The clinical use of human culture-expanded autologous bone marrow mesenchymal stem cells transplanted on platelet-rich fibrin glue in the treatment of articular cartilage defects: a pilot study and preliminary results. Cartilage. 2010;1(4):253–61.

  28. 28.

    Koga H, Muneta T, Ju YJ, Nagase T, Nimura A, Mochizuki T, et al. Synovial stem cells are regionally specified according to local microenvironments after implantation for cartilage regeneration. Stem Cells. 2007;25(3):689–96.

  29. 29.

    Mazaki T, Shiozaki Y, Yamane K, Yoshida A, Nakamura M, Yoshida Y, et al. A novel, visible light-induced, rapidly cross-linkable gelatin scaffold for osteochondral tissue engineering. Sci Rep. 2014;4:4457.

  30. 30.

    Miller RE, Grodzinsky AJ, Vanderploeg EJ, Lee C, Ferris DJ, Barrett MF, et al. Effect of self-assembling peptide, chondrogenic factors, and bone marrow-derived stromal cells on osteochondral repair. Osteoarthr Cartil. 2010;18(12):1608–19.

  31. 31.

    D’Este M, Sprecher CM, Milz S, Nehrbass D, Dresing I, Zeiter S, et al. Evaluation of an injectable thermoresponsive hyaluronan hydrogel in a rabbit osteochondral defect model. J Biomed Mater Res A. 2016;104(6):1469–78.

  32. 32.

    Levingstone TJ, Thompson E, Matsiko A, Schepens A, Gleeson JP, O’Brien FJ. Multi-layered collagen-based scaffolds for osteochondral defect repair in rabbits. Acta Biomater. 2016;32:149–60.

  33. 33.

    Pulkkinen HJ, Tiitu V, Valonen P, Jurvelin JS, Rieppo L, Töyräs J, et al. Repair of osteochondral defects with recombinant human type II collagen gel and autologous chondrocytes in rabbit. Osteoarthr Cartil. 2013;21(3):481–90.

  34. 34.

    Liu S, Jia Y, Yuan M, Guo W, Huang J, Zhao B, et al. Repair of osteochondral defects using human umbilical cord Wharton’s jelly-derived mesenchymal stem cells in a rabbit model. BioMed Res Int. 2017;2017:12.

  35. 35.

    Jang K-M, Lee J-H, Park CM, Song H-R, Wang JH. Xenotransplantation of human mesenchymal stem cells for repair of osteochondral defects in rabbits using osteochondral biphasic composite constructs. Knee Surg Sports Traumatol Arthrosc. 2014;22(6):1434–44.

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Acknowledgements

The authors are grateful for funding from the European Commission (NMP-2014-646075, HEALTH-F4-2011-278557, PITN-GA-2012-317306 and MSCA-ITN-2014-642687), the MINECO of the Spanish Government (MAT2016-78903-R, MAT2016-79435-R, MAT2013-42473-R, MAT2013-41723-R and MAT2012-38043), the Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, and Junta de Castilla y León (VA244U13, VA313U14 and GRS/516/A/10), Spain. Sandra Muntión is supported by grant RD12/0019/0017 from the Instituto de Salud Carlos III, Spain.

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Correspondence to José Carlos Rodríguez-Cabello.

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Pescador, D., Ibáñez-Fonseca, A., Sánchez-Guijo, F. et al. Regeneration of hyaline cartilage promoted by xenogeneic mesenchymal stromal cells embedded within elastin-like recombinamer-based bioactive hydrogels. J Mater Sci: Mater Med 28, 115 (2017). https://doi.org/10.1007/s10856-017-5928-1

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