Cryopreservation does not alter the ultrastructure of the meniscus

  • Pablo Eduardo GelberEmail author
  • Gemma Gonzalez
  • Raúl Torres
  • Natalia Garcia Giralt
  • Enrique Caceres
  • Juan Carlos Monllau


Fresh frozen menisci have recently been shown to have an altered meniscal ultrastructure. The cause might be a deterioration of its permeability due to collagen net disarray. The purpose of this study was to evaluate the cryopreserved meniscus in terms of ultrastructure and cellularity. Ten fresh human lateral menisci were harvested. Collagen architecture was evaluated with transmission electron microscopy. The Collagen Meniscal Architecture scoring system was used to assess the degree of meniscal disarray. Cell population, was also evaluated. The fibril collagen diameters of those menisci which had been previously cryopreserved showed an average size in the longitudinal section of 12.6 ± 1.3 nm, whereas it was 13.4 ± 2.2 nm in the menisci used as controls (n.s.). In the transverse section, the cryopreserved menisci averaged 15.5 ± 2.4 and 16.7 ± 3.5 nm in the controls (n.s.). The study group scored 4.8 points ± 1.7, whereas the control group did so at 4.1 ± 1.3 (n.s.). The percentage of cell survival after the cryopreservation ranged from 4 to 54. The fibril diameters and degree of disarray showed a similar distribution in both groups. The results suggest that meniscal cryopreservation does not alter the meniscal ultrastructure. Therefore, an allograft stored in that way would not alter its biomechanical properties, although its cellular viability is highly unpredictable.


Meniscus allograft Shrinkage Meniscal nutrition Transmission electron microscopy CMA scoring system 


  1. 1.
    Arnoczky SP, McDevitt CA, Schmidt MB, Mow VC, Warren RF (1988) The effect of cryopreservation on canine menisci: a biochemical, morphologic, and biomechanical evaluation. J Orthop Res 6:1–12PubMedCrossRefGoogle Scholar
  2. 2.
    Arnoczky SP, Dicarlo EF, O’Brien SJ, Warren RF (1992) Cellular repopulation of deep-frozen meniscal autografts: an experimental study in the dog. Arthroscopy 8:428–436PubMedGoogle Scholar
  3. 3.
    Chen J, Iosifidis M, Zhu J, Tatarinsev I, Wang J (2006) Vanadate ingestion enhances the organization and collagen fibril diameters of rat healing medical collateral ligaments. Knee Surg Sports Traumatol Arthrosc 14:750–755PubMedCrossRefGoogle Scholar
  4. 4.
    DeBeer P, Decorte R, Delvaux S, Bellemans J (2000) DNA analysis of a transplanted cryopreserved meniscal allograft. Arthroscopy 16:71–75PubMedGoogle Scholar
  5. 5.
    Fabbriciani C, Lucania L, Milano G, Schiavone Panni A, Evangelisti M (1997) Meniscal allografts: cryopreservation versus deep-frozen technique. An experimental study in goats. Knee Surg Sports Traumatol Arthrosc 5:124–134PubMedCrossRefGoogle Scholar
  6. 6.
    Gelber PE, Gonzalez G, Lloreta JL, Reina F, Caceres E, Monllau JC (2008) Freezing causes changes in the meniscus collagen net: a new ultrastructural meniscus disarray scale. Knee Surg Sports Traumatol Arthrosc 16:353–359PubMedCrossRefGoogle Scholar
  7. 7.
    Giannini S, Buda R, Di Caprio F, Agati P, Bigi A, De Pascuale V, Ruggeri A (2008) Effects of freezing on the biomechanical and structural properties of human posterior tibial tendons. Int Orthop 32:145–151PubMedCrossRefGoogle Scholar
  8. 8.
    Graf KW Jr, Sekiya JK, Wojtys EM (2004) Long-term results after combined medial meniscal allograft transplantation and anterior cruciate ligament reconstruction: minimum 8.5-year follow-up study. Arthroscopy 20:129–140PubMedGoogle Scholar
  9. 9.
    Hommen JP, Applegate GR, Del Pizzo W (2007) Meniscus allograft transplantation: ten year results of cryopreserved allografts. Arthroscopy 4:388–393Google Scholar
  10. 10.
    Lewis PB, Williams JM, Hallab N, Virdi A, Yanke A, Cole BJ (2008) Multiple freeze–thaw cycled meniscal allograft tissue: a biomechanical, biochemical, and histologic analysis. J Orthop Res 1:49–55CrossRefGoogle Scholar
  11. 11.
    Noyes FR, Barber-Westin SD, Butler DL, Wilkins RM (1998) The role of allografts in repair and reconstruction of knee joint ligaments and menisci. Instr Course Lect 47:379–396PubMedGoogle Scholar
  12. 12.
    Ochi M, Kanda T, Sumen Y, Ikuta Y (1997) Changes in the permeability and histologic findings of rabbit menisci after immobilization. Clin Orthop Relat Res 334:305–315PubMedCrossRefGoogle Scholar
  13. 13.
    Pegg DE (2006) The preservation of tissues for transplantation. Cell Tissue Bank 7:349–358PubMedCrossRefGoogle Scholar
  14. 14.
    Rodeo SA, Seneviratne A, Suzuki K, Felker K, Wickiewicz TL, Warren RF (2000) Histological analysis of human meniscal allografts, a preliminary report. J Bone Joint Surg Am 82-A:1071–1082PubMedGoogle Scholar
  15. 15.
    Salai M, Givon U, Messer Y, von Versen R (1997) Electron microscopic study on the effects of different preservation methods for meniscal cartilage. Ann Transplant 2:52–54PubMedGoogle Scholar
  16. 16.
    Shibuya S (1999) Meniscus transplantation using a cryopreserved allograft. Histological and ultrastructural study of the transplanted meniscus. J Orthop Sci 4:135–141PubMedCrossRefGoogle Scholar
  17. 17.
    Sumida S (2006) Transfusion and transplantation of cryopreserved cells and tissues. Cell Tissue Bank 7:265–305PubMedCrossRefGoogle Scholar
  18. 18.
    Verdonk PCM, Demurie A, Almqvist KF, Veys EM, Verbruggen G, Verdonk R (2005) Transplantation of viable meniscal allograft: survivorship analysis and clinical outcome of one hundred cases. J Bone Joint Surg Am 87:715–724PubMedCrossRefGoogle Scholar
  19. 19.
    Verdonk PCM, Verstraete KL, Almqvist KF, De Cuyper K, Veys EM, Verbruggen G, Verdonk R (2006) Meniscal allograft transplantation: long-term clinical results with radiological and magnetic imaging correlations. Knee Surg Sports Traumatol Arthrosc 14:694–706PubMedCrossRefGoogle Scholar
  20. 20.
    Verdonk R, Khon D (1999) Harvest and conservation of meniscal allografts. Scand J Med Sci Sports 9:158–159PubMedCrossRefGoogle Scholar
  21. 21.
    Wada Y, Amiel M, Harwood F, Moriya H, Amiel D (1998) Architectural remodeling in deep frozen meniscal allografts after total meniscectomy. Arthroscopy 14:250–257PubMedGoogle Scholar
  22. 22.
    Wirth CJ, Peters G, Milachowski KA, Weismeier KG, Kohn D (2002) Long-term results of meniscal allograft transplantation. Am J Sports Med 30:174–181PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Pablo Eduardo Gelber
    • 1
    Email author
  • Gemma Gonzalez
    • 1
  • Raúl Torres
    • 1
  • Natalia Garcia Giralt
    • 1
  • Enrique Caceres
    • 1
  • Juan Carlos Monllau
    • 1
  1. 1.Department of Orthopaedic Surgery, URFOA-IMIM, Hospital Universitari del MarUniversitat Autònoma de BarcelonaBarcelonaSpain

Personalised recommendations