Gene Therapy, Growth Factors, Mesenchymal Cells, New Trends and Future Perspectives

  • Stefano Zaffagnini
  • Magali Cucchiarini
  • Laura de Girolamo
  • Peter Angele
  • Helder Pereira
  • Rui L. Reis
  • Henning Madry
  • Carlotta Perucca Orfei
  • Johannes Zellner
  • Giuseppe Filardo
Chapter

Abstract

Meniscal tears are common, prevalent intra-articular knee injury and the most frequent cause of orthopedic surgical procedures [1], being a significant risk factor for the development of osteoarthritis (OA) [2]. As widely documented in the previous chapters, tears in the peripheral (vascularized) portion of the meniscus can be repaired using a variety of operative procedures, while those in the central (avascular) area have a poorer healing capacity. Reconstruction of a torn meniscus in this location is challenging, and the long-term effect of allografts on the progression of OA remains uncertain [3].

Notes

Acknowledgment

Parts of this chapter have been published by Angele et al., JEO1 (2014).

References

  1. 1.
    Beaufils P, Hulet C, Dhenain M, Nizard R, Nourissat G, Pujol N (2009) Clinical practice guidelines for the management of meniscal lesions and isolated lesions of the anterior cruciate ligament of the knee in adults. Orthop Traumatol Surg Res 95(6):437–442CrossRefPubMedGoogle Scholar
  2. 2.
    Salata MJ, Gibbs AE, Sekiya JK (2010) A systematic review of clinical outcomes in patients undergoing meniscectomy. Am J Sports Med 38(9):1907–1916CrossRefPubMedGoogle Scholar
  3. 3.
    Rosso F, Bisicchia S, Bonasia DE, Amendola A (2015) Meniscal allograft transplantation: a systematic review. Am J Sports Med 43(4):998–1007CrossRefPubMedGoogle Scholar
  4. 4.
    Lee HP, Kaul G, Cucchiarini M, Madry H (2014) Nonviral gene transfer to human meniscal cells. Part I: transfection analyses and cell transplantation to meniscus explants. Int Orthop 38(9):1923–1930CrossRefPubMedGoogle Scholar
  5. 5.
    Evans CH, Huard J (2015) Gene therapy approaches to regenerating the musculoskeletal system. Nat Rev Rheumatol 11(4):234–242CrossRefPubMedGoogle Scholar
  6. 6.
    Cucchiarini M, Madry H (2005) Gene therapy for cartilage defects. J Gene Med 7(12):1495–1509CrossRefPubMedGoogle Scholar
  7. 7.
    Wang ZH, Li XL, He XJ, Wu BJ, Xu M, Chang HM, Yang YY (2014) Delivery of the Sox9 gene promotes chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in an in vitro model. Braz J Med Biol Res 47(4):279–286CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Garza-Veloz I, Romero-Diaz VJ, Martinez-Fierro ML, Marino-Martinez IA, Gonzalez-Rodriguez M, Martinez-Rodriguez HG, Rojas-Martinez A (2013) Analyses of chondrogenic induction of adipose mesenchymal stem cells by combined co-stimulation mediated by adenoviral gene transfer. Arthritis Res Ther 15(4):R80–R91CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Martinek V, Usas A, Pelinkovic D, Robbins P, Fu FH, Huard J (2002) Genetic engineering of meniscal allografts. Tissue Eng 8(1):107–117CrossRefPubMedGoogle Scholar
  10. 10.
    Brunger JM, Huynh NP, Guenther CM, Perez-Pinera P, Moutos FT, Sanchez-Adams J, Guilak F (2014) Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage. Proc Natl Acad Sci U S A 111(9):E798–E806CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Gerich TG, Ghivizani S, Fu FH, Robbins PD, Evans CH (1997) Gene transfer into the patellar tendon of rabbits: a preliminary study of locoregional expression of growth factors. Wien Klin Wochenschr 109(11):384–389PubMedGoogle Scholar
  12. 12.
    Cucchiarini M, Schmidt K, Frisch J, Kohn D, Madry H (2015) Overexpression of TGF-beta via rAAV-mediated gene transfer promotes the healing of human meniscal lesions ex vivo on explanted menisci. Am J Sports Med 43(5):1197–1205CrossRefPubMedGoogle Scholar
  13. 13.
    Cucchiarini M, Schetting S, Terwilliger EF, Kohn D, Madry H (2009) rAAV-mediated overexpression of FGF-2 promotes cell proliferation, survival, and alpha-SMA expression in human meniscal lesions. Gene Ther 16(11):1363–1372CrossRefPubMedGoogle Scholar
  14. 14.
    Goto H, Shuler FD, Lamsam C, Moller HD, Niyibizi C, Fu FH, Evans CH (1999) Transfer of lacZ marker gene to the meniscus. J Bone Joint Surg Am 81(7):918–925PubMedGoogle Scholar
  15. 15.
    Nakata K, Shino K, Hamada M, Mae T, Miyama T, Shinjo H, Yoshikawa H (2001) Human meniscus cell: characterization of the primary culture and use for tissue engineering. Clin Orthop Relat Res 391(Suppl):S208–S218Google Scholar
  16. 16.
    Steinert AF, Palmer GD, Capito R, Hofstaetter JG, Pilapil C, Ghivizzani SC, Evans CH (2007) Genetically enhanced engineering of meniscus tissue using ex vivo delivery of transforming growth factor-beta 1 complementary deoxyribonucleic acid. Tissue Eng 13(9):2227–2237CrossRefPubMedGoogle Scholar
  17. 17.
    Hidaka C, Ibarra C, Hannafin JA, Torzilli PA, Quitoriano M, Jen SS, Crystal RG (2002) Formation of vascularized meniscal tissue by combining gene therapy with tissue engineering. Tissue Eng 8(1):93–105CrossRefPubMedGoogle Scholar
  18. 18.
    Zhang H, Leng P, Zhang J (2009) Enhanced meniscal repair by overexpression of hIGF-1 in a full-thickness model. Clin Orthop Relat Res 467(12):3165–3174CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Huang G, Zheng Q, Sun J, Guo C, Yang J, Chen R, Wang J (2008) Stabilization of cellular properties and differentiation multipotential of human mesenchymal stem cells transduced with hTERT gene in a long-term culture. J Cell Biochem 103(4):1256–1269CrossRefPubMedGoogle Scholar
  20. 20.
    Filardo G, Kon E, Roffi A et al (2013) Platelet-rich plasma: why intra-articular? A systematic review of preclinical studies and clinical evidence on PRP for joint degeneration. Knee Surg Sports Traumatol Arthrosc. http://link.springer.com/article/10.1007/s00167-013-2743-1
  21. 21.
    Volpi P, Quaglia A, Schoenhuber H et al (2010) Growth factors in the management of sport-induced tendinopathies: results after 24 months from treatment. A pilot study. J Sports Med Phys Fitness 50:494–500PubMedGoogle Scholar
  22. 22.
    Tschon M, Fini M, Giardino R, Filardo G, Dallari D, Torricelli P, Martini L, Giavaresi G, Kon E, Maltarello MC, Nicolini A, Carpi A (2011) Lights and shadows concerning platelet products for musculoskeletal regeneration. Front Biosci (Elite Ed) 3:96–107CrossRefGoogle Scholar
  23. 23.
    Di Matteo B, Filardo G, Kon E et al (2015) Platelet-rich plasma: evidence for the treatment of patellar and Achilles tendinopathy-a systematic review. Musculoskelet Surg 99:1–9CrossRefPubMedGoogle Scholar
  24. 24.
    Griffin JW, Hadeed MM, Werner BC, Diduch DR, Carson EW, Miller MD (2015) Platelet-rich plasma in meniscal repair: does augmentation improve surgical outcomes? Clin Orthop Relat Res 473(5):1665–1672CrossRefPubMedGoogle Scholar
  25. 25.
    Pujol N, Salle De Chou E, Boisrenoult P, Beaufils P (2015) Platelet-rich plasma for open meniscal repair in young patients: any benefit? Knee Surg Sports Traumatol Arthrosc 23:51–58CrossRefPubMedGoogle Scholar
  26. 26.
    Ishida K, Kuroda R, Miwa M et al (2007) The regenerative effects of platelet rich plasma on meniscal cells in vitro and its in vivo application with biodegradable gelatin hydrogel. Tissue Eng 13:1103–1112CrossRefPubMedGoogle Scholar
  27. 27.
    Kon E, Filardo G, Tschon M, Fini M, Giavaresi G, Marchesini Reggiani L, Chiari C, Nehrer S, Martin I, Salter DM, Ambrosio L, Marcacci M (2012) Tissue engineering for total meniscal substitution: animal study in sheep model – results at 12 months. Tissue Eng Part A 18(15–16):1573–1582CrossRefPubMedGoogle Scholar
  28. 28.
    Kwak HS, Nam J, Lee JH, Kim HJ, Yoo JJ (2014) Meniscal repair in vivo using human chondrocyte-seeded PLGA mesh scaffold pretreated with platelet-rich plasma. J Tissue Eng Regen Med [Epub ahead of print]. doi:10.1002/term.1938
  29. 29.
    Zellner J, Taeger CD, Schaffer M, Roldan JC, Loibl M, Mueller MB, Berner A, Krutsch W, Huber MK, Kujat R, Nerlich M, Angele P (2014) Are applied growth factors able to mimic the positive effects of mesenchymal stem cells on the regeneration of meniscus in the avascular zone? Biomed Res Int 2014:537686CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Scotti C, Hirschmann MT, Antinolfi P, Martin I, Peretti GM (2013) Meniscus repair and regeneration: review on current methods and research potential. Eur Cell Mater 26:150–170PubMedGoogle Scholar
  31. 31.
    Freymann U, Endres M, Goldmann U, Sittinger M, Kaps C (2013) Toward scaffold-based meniscus repair: effect of human serum, hyaluronic acid and TGF-ß3 on cell recruitment and re-differentiation. Osteoarthritis Cartilage 21:773–781CrossRefPubMedGoogle Scholar
  32. 32.
    Esparza R, Gortazar AR, Forriol F (2012) Cell study of the three areas of the meniscus: effect of growth factors in an experimental model in sheep. J Orthop Res 30:1647–1651CrossRefPubMedGoogle Scholar
  33. 33.
    Caplan AI, Dennis JE (2006) Mesenchymal stem cells as trophic mediators. J Cell Biochem 98(5):1076–1084. doi:10.1002/jcb.20886 CrossRefPubMedGoogle Scholar
  34. 34.
    Arnoczky SP (1999) Building a meniscus. Biologic considerations. Clin Orthop Relat Res 367(Suppl):S244–253Google Scholar
  35. 35.
    Filardo G, Madry H, Jelic M, Roffi A, Cucchiarini M, Kon E (2013) Mesenchymal stem cells for the treatment of cartilage lesions: from preclinical findings to clinical application in orthopaedics. Knee Surg Sports Traumatol Arthrosc 21(8):1717–1729CrossRefPubMedGoogle Scholar
  36. 36.
    Crisan M, Corselli M, Chen CW, Peault B (2011) Multilineage stem cells in the adult: a perivascular legacy? Organogenesis 7(2):101–104CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Angele P, Kujat R, Koch M, Zellner J (2014) Role of mesenchymal stem cells in meniscal repair. J Exp Orthop 1:12CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Dowthwaite GP, Bishop JC, Redman SN, Khan IM, Rooney P, Evans DJ, Haughton L, Bayram Z, Boyer S, Thomson B, Wolfe MS, Archer CW (2004) The surface of articular cartilage contains a progenitor cell population. J Cell Sci 117(Pt 6):889–897. doi:10.1242/jcs.00912 CrossRefPubMedGoogle Scholar
  39. 39.
    Mauck RL, Martinez-Diaz GJ, Yuan X, Tuan RS (2007) Regional multilineage differentiation potential of meniscal fibrochondrocytes: implications for meniscus repair. Anat Rec 290(1):48–58. doi:10.1002/ar.20419 CrossRefGoogle Scholar
  40. 40.
    McCarthy HE, Bara JJ, Brakspear K, Singhrao SK, Archer CW (2012) The comparison of equine articular cartilage progenitor cells and bone marrow-derived stromal cells as potential cell sources for cartilage repair in the horse. Vet J 192(3):345–351. doi:10.1016/j.tvjl.2011.08.036 CrossRefPubMedGoogle Scholar
  41. 41.
    Matsukura Y, Muneta T, Tsuji K, Koga H, Sekiya I (2014) Mesenchymal stem cells in synovial fluid increase after meniscus injury. Clin Orthop Relat Res 472(5):1357–1364. doi:10.1007/s11999-013-3418-4 CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Horie M, Sekiya I, Muneta T, Ichinose S, Matsumoto K, Saito H, Murakami T, Kobayashi E (2009) Intra-articular Injected synovial stem cells differentiate into meniscal cells directly and promote meniscal regeneration without mobilization to distant organs in rat massive meniscal defect. Stem Cells 27(4):878–887. doi:10.1634/stemcells.2008-0616 CrossRefPubMedGoogle Scholar
  43. 43.
    Nepple JJ, Dunn WR, Wright RW (2012) Meniscal repair outcomes at greater than five years: a systematic literature review and meta-analysis. J Bone Joint Surg Am 94(24):2222–2227. doi:10.2106/jbjs.k.01584 CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Zellner J, Hierl K, Mueller M, Pfeifer C, Berner A, Dienstknecht T, Krutsch W, Geis S, Gehmert S, Kujat R, Dendorfer S, Prantl L, Nerlich M, Angele P (2013) Stem cell-based tissue-engineering for treatment of meniscal tears in the avascular zone. J Biomed Mater Res B Appl Biomater 101(7):1133–1142. doi:10.1002/jbm.b.32922 CrossRefPubMedGoogle Scholar
  45. 45.
    Vangsness CT Jr, Farr J 2nd, Boyd J, Dellaero DT, Mills CR, LeRoux-Williams M (2014) Adult human mesenchymal stem cells delivered via intra-articular injection to the knee following partial medial meniscectomy: a randomized, double-blind, controlled study. J Bone Joint Surg Am 96(2):90–98. doi:10.2106/JBJS.M.00058 CrossRefPubMedGoogle Scholar
  46. 46.
    Filardo G, Andriolo L, Kon E, de Caro F, Marcacci M (2015) Meniscal scaffolds: results and indications. A systematic literature review. Int Orthop 39(1):35–46CrossRefPubMedGoogle Scholar
  47. 47.
    Di Matteo B, Perdisa F, Gostynska N, Kon E, Filardo G, Marcacci M (2015) Meniscal scaffolds – preclinical evidence to support their use: a systematic review. Open Orthop J 9:143–156. doi:10.2174/1874325001509010143 CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Kon E, Filardo G, Zaffagnini S, Di Martino A, Di Matteo B, Marcheggiani Muccioli GM, Busacca M, Marcacci M (2014) Biodegradable polyurethane meniscal scaffold for isolated partial lesions or as combined procedure for knees with multiple comorbidities: clinical results at 2 years. Knee Surg Sports Traumatol Arthrosc 22(1):128–134CrossRefPubMedGoogle Scholar
  49. 49.
    Zaffagnini S, Marcheggiani Muccioli GM, Bulgheroni P, Bulgheroni E, Grassi A, Bonanzinga T, Kon E, Filardo G, Busacca M, Marcacci M (2012) Arthroscopic collagen meniscus implantation for partial lateral meniscal defects: a 2-year minimum follow-up study. Am J Sports Med 40(10):2281–2288CrossRefPubMedGoogle Scholar
  50. 50.
    Ishimura M, Tamai S, Fujisawa Y (1991) Arthroscopic meniscal repair with fibrin glue. Arthroscopy 7(2):177–181CrossRefPubMedGoogle Scholar
  51. 51.
    Angele P, Muller R, Schumann D, Englert C, Zellner J, Johnstone B, Yoo J, Hammer J, Fierlbeck J, Angele MK, Nerlich M, Kujat R (2009) Characterization of esterified hyaluronan-gelatin polymer composites suitable for chondrogenic differentiation of mesenchymal stem cells. J Biomed Mater Res A 91(2):416–427. doi:10.1002/jbm.a.32236 CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Pereira H, Silva-Correia J, Oliveira JM, Reis RL, Espregueira-Mendes J (2013) Future trends in the treatment of meniscus lesions: from repair to regeneration. In: Verdonk R, Espregueira-Mendes J, Monllau JC (eds) Meniscal transplantation. Springer, Heidelberg/New York/Dordrecht/London, pp 103–114CrossRefGoogle Scholar
  53. 53.
    Pereira H, Frias AM, Oliveira JM, Espregueira-Mendes J, Reis RL (2011) Tissue engineering and regenerative medicine strategies in meniscus lesions. Arthroscopy 27:1706–1719CrossRefPubMedGoogle Scholar
  54. 54.
    Yan LP, Oliveira JM, Oliveira AL, Caridade SG, Mano JF, Reis RL (2012) Macro/microporous silk fibroin scaffolds with potential for articular cartilage and meniscus tissue engineering applications. Acta Biomater 8:289–301CrossRefPubMedGoogle Scholar
  55. 55.
    Welsing RT, van Tienen TG, Ramrattan N, Heijkants R, Schouten AJ, Veth RP et al (2008) Effect on tissue differentiation and articular cartilage degradation of a polymer meniscus implant: a 2-year follow-up study in dogs. Am J Sports Med 36:1978–1989CrossRefPubMedGoogle Scholar
  56. 56.
    Kang SW, Son SM, Lee JS, Lee ES, Lee KY, Park SG et al (2006) Regeneration of whole meniscus using meniscal cells and polymer scaffolds in a rabbit total meniscectomy model. J Biomed Mater Res A 78:659–671CrossRefPubMedGoogle Scholar
  57. 57.
    Kim IL, Mauck RL, Burdick JA (2011) Hydrogel design for cartilage tissue engineering: a case study with hyaluronic acid. Biomaterials 32:8771–8782CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Heo J, Koh RH, Shim W, Kim HD, Yim HG, Hwang NS (2015) Riboflavin-induced photo-crosslinking of collagen hydrogel and its application in meniscus tissue engineering. Drug Deliv Transl Res [Epub ahead of print]Google Scholar
  59. 59.
    Silva-Correia J, Gloria A, Oliveira MB, Mano JF, Oliveira JM, Ambrosio L et al (2013) Rheological and mechanical properties of acellular and cell-laden methacrylated gellan gum hydrogels. J Biomed Mater Res A 101:3438–3446CrossRefPubMedGoogle Scholar
  60. 60.
    Huey DJ, Athanasiou KA (2011) Maturational growth of self-assembled, functional menisci as a result of TGF-β1 and enzymatic chondroitinase-ABC stimulation. Biomaterials 32:2052–2058CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    MacBarb RF, Makris EA, Hu JC, Athanasiou KA (2012) A chondroitinase-ABC and TGF-β1 treatment regimen for enhancing the mechanical properties of tissue engineered fibrocartilage. Acta Biomater 9(1):4626–4634CrossRefPubMedPubMedCentralGoogle Scholar
  62. 62.
    Ionescu LC, Lee GC, Huang KL, Mauck RL (2012) Growth factor supplementation improves native and engineered meniscus repair in vitro. Acta Biomater 8:3687–3694CrossRefPubMedPubMedCentralGoogle Scholar
  63. 63.
    Gu Y, Wang Y, Dai H, Lu L, Cheng Y, Zhu W (2012) Chondrogenic differentiation of canine myoblasts induced by cartilage-derived morphogenetic protein-2 and transforming growth factor-β1 in vitro. Mol Med Rep 5:767–772PubMedGoogle Scholar
  64. 64.
    Hoben GM, Willard VP, Athanasiou KA (2009) Fibrochondrogenesis of hESCs: growth factor combinations and cocultures. Stem Cells Dev 18:283–292CrossRefPubMedPubMedCentralGoogle Scholar
  65. 65.
    Pörtner R, Nagel-Heyer S, Goepfert C, Adamietz P, Meenen NM (2005) Bioreactor design for tissue engineering. J Biosci Bioeng 100:235–245CrossRefPubMedGoogle Scholar
  66. 66.
    Petri M, Ufer K, Toma I, Becher C, Liodakis E, Brand S et al (2012) Effects of perfusion and cyclic compression on in vitro tissue engineered meniscus implants. Knee Surg Sports Traumatol Arthrosc 20:223–231CrossRefPubMedGoogle Scholar
  67. 67.
    Martínez H, Brackmann C, Enejder A, Gatenholm P (2012) Mechanical stimulation of fibroblasts in micro‐channeled bacterial cellulose scaffolds enhances production of oriented collagen fibers. J Biomed Mater Res A 100:948–957CrossRefPubMedGoogle Scholar
  68. 68.
    Fox DB, Warnock JJ, Stoker AM, Luther JK, Cockrell M (2010) Effects of growth factors on equine synovial fibroblasts seeded on synthetic scaffolds for avascular meniscal tissue engineering. Res Vet Sci 88:326–332CrossRefPubMedGoogle Scholar
  69. 69.
    Gunja NJ, Dujari D, Chen A, Luengo A, Fong JV, Hung CT (2012) Migration responses of outer and inner meniscus cells to applied direct current electric fields. J Orthop Res 30:103–111CrossRefPubMedPubMedCentralGoogle Scholar
  70. 70.
    Pasa L, Visna P (2005) Suture of meniscus. Scripta Medica. (BRNO), 78(3):135–150Google Scholar
  71. 71.
    Pereira H, Caridade SG, Frias AM, Silva-Correia J, Pereira DR, Cengiz IF et al (2014) Biomechanical and cellular segmental characterization of human meniscus: building the basis for tissue engineering therapies. Osteoarthritis Cartilage 22:1271–1281CrossRefPubMedGoogle Scholar
  72. 72.
    Makris EA, Hadidi P, Athanasiou KA (2011) The knee meniscus: structure-function, pathophysiology, current repair techniques, and prospects for regeneration. Biomaterials 32:7411–7431CrossRefPubMedPubMedCentralGoogle Scholar
  73. 73.
    Goto H, Shuler FD, Niyibizi C, Fu FH, Robbins PD,Evans CH (2000) Gene therapy for meniscal injury: enhanced synthesis of proteoglycan and collagen by meniscal cells transduced with a TGFbeta(1)gene. Osteoarthritis Cartilage 8(4):266–271Google Scholar
  74. 74.
    Simonsen JL, Rosada C, Serakinci N, Justesen J, Stenderup K, Rattan SI, Kassem M (2002) Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells. Nat Biotechnol 20(6):592–596Google Scholar
  75. 75.
    Cucchiarini M, Ekici M, Schetting S, Kohn D, Madry H (2011) Metabolic activities and chondrogenic differentiation of human mesenchymal stem cells following recombinant adeno-associated virus-mediated gene transfer and overexpression of fibroblast growth factor 2. Tissue Eng Part A 17(15–16):1921–1933Google Scholar

Copyright information

© ESSKA 2016

Authors and Affiliations

  • Stefano Zaffagnini
    • 1
  • Magali Cucchiarini
    • 2
  • Laura de Girolamo
    • 3
  • Peter Angele
    • 4
    • 5
  • Helder Pereira
    • 6
    • 7
    • 8
    • 9
  • Rui L. Reis
    • 10
    • 8
  • Henning Madry
    • 2
    • 11
  • Carlotta Perucca Orfei
    • 3
  • Johannes Zellner
    • 4
  • Giuseppe Filardo
    • 1
  1. 1.II Clinic – Biomechanics and Innovation Technology LaboratoryRizzoli Orthopaedic InstituteBolognaItaly
  2. 2.Center of Experimental OrthopaedicsSaarland University Medical CenterHomburg/SaarGermany
  3. 3.Orthopaedic Biotechnology LaboratoryIRCCS Galeazzi Orthopaedic InstituteMilanItaly
  4. 4.Department of Trauma SurgeryUniversity Medical Center RegensburgRegensburgGermany
  5. 5.SporthopaedicumRegensburgGermany
  6. 6.Orthopedic DepartmentCentro Hospitalar Póvoa de Varzim - Vila do CondeVila do CondePortugal
  7. 7.3B’s Research Group – Biomaterials, Biodegradables and BiomimeticsUniv. Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative MedicineGuimarãesPortugal
  8. 8.ICVS/3B’s – PT Government Associated LaboratoryBragaPortugal
  9. 9.Ripoll y De Prado Sports Clinic FIFA Medical Centre of ExcellenceMurcia-MadridSpain
  10. 10.Tissue Engineering and Regenerative Medicine, 3B’s Research Group – Biomaterials, Biodegradables and BiomimeticsUniversity Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative MedicineGuimarãesPortugal
  11. 11.Department of Orthopaedic SurgerySaarland University Medical CenterHomburg/SaarGermany

Personalised recommendations