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J Knee Surg 2012; 25(02): 117-126
DOI: 10.1055/s-0032-1319809
Special Focus Section
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Improved Preservation of Fresh Osteochondral Allografts for Clinical Use

Aaron Stoker
1   Department of Veterinary Medicine and Surgery, University of Missouri, Columbia, Missouri
2   Comparative Orthopaedic Laboratory, University of Missouri, Columbia, Missouri
,
Joeseph T. Garrity
2   Comparative Orthopaedic Laboratory, University of Missouri, Columbia, Missouri
,
Clark T. Hung
3   Department of Biomedical Engineering, Columbia University, New York, New York
,
James P. Stannard
4   Department of Orthopaedic Surgery, University of Missouri Hospital, Columbia, Missouri
,
Jimi Cook
2   Comparative Orthopaedic Laboratory, University of Missouri, Columbia, Missouri
› Institutsangaben
Weitere Informationen

Publikationsverlauf

31. Oktober 2011

16. Mai 2012

Publikationsdatum:
28. Juni 2012 (online)

    Lizenzen und Reprints

Abstract

Introduction Fresh osteochondral allografts (OCAs) have been used clinically to treat cartilage focal defects of the knee for over 30 years. Over the last decade, significant research has been performed to develop and improve protocols for preservation of osteochondral tissue before transplantation into patients for treatment of cartilage defects. This work has resulted in preservation protocols that allow for maintenance of OCA tissues for time periods sufficient for clinical use based on disease testing requirements in the United States. However, graft quality and the window for clinical use of these tissues could be greatly enhanced from current levels.

Materials and Methods Femoral condyles from 14 dogs were harvested and stored in one of the three proprietary media composition (M-1, M-2, M-3) and container condition (C-1, C-2, C-3) at 25°C for 63 days. Viability of the OCA was determined using a proprietary media metabolic assay and live cell fluorescent microscopy. Media biomarker concentrations were analyzed to determine the metabolic activity of tissue.

Results Media protein biomarkers were detected throughout the culture period, indicating OCAs remain metabolically active at 25°C, and biomarker levels correlated with tissue viability. Viable chondrocyte density was maintained at day 0 levels throughout the depth of the tissue in the M-3 media using container condition C-3 after 63 days in storage. The media metabolic assay correlated strongly to cell viability of the OCA tissue.

Conclusion These data indicate that near day 0 tissue viability can be maintained for up to 63 days when OCAs are stored at 25°C in the correct conditions. Further, tissue viability could be assessed nondestructively using media biomarkers and the media metabolic assay. If the preservation protocol reported here can be validated for safety and functional outcome, it could then be employed in tissue banks throughout the world, decreasing the number of grafts discarded and improving quality of life for thousands of patients affected by cartilage defects.

Keywords

osteochondral allograft - osteoarthritis - storage - viability
 

  • References

  • 1 Gross AE, Silverstein EA, Falk J, Falk R, Langer F. The allotransplantation of partial joints in the treatment of osteoarthritis of the knee. Clin Orthop Relat Res 1975; (108) 7-14
    PubMedGoogle Scholar
  • 2 Williams III RJ, Ranawat AS, Potter HG, Carter T, Warren RF. Fresh stored allografts for the treatment of osteochondral defects of the knee. J Bone Joint Surg Am 2007; 89 (4) 718-726
    CrossrefPubMedGoogle Scholar
  • 3 Gross AE, Kim W, Las Heras F, Backstein D, Safir O, Pritzker KP. Fresh osteochondral allografts for posttraumatic knee defects: long-term followup. Clin Orthop Relat Res 2008; 466 (8) 1863-1870
    Google Scholar
  • 4 Farr J, Cole B, Dhawan A, Kercher J, Sherman S. Clinical cartilage restoration: evolution and overview. Clin Orthop Relat Res 2011; 469 (10) 2696-2705
    CrossrefPubMedGoogle Scholar
  • 5 Bakay A, Csönge L, Papp G, Fekete L. Osteochondral resurfacing of the knee joint with allograft. Clinical analysis of 33 cases. Int Orthop 1998; 22 (5) 277-281
    CrossrefPubMedGoogle Scholar
  • 6 Beaver RJ, Mahomed M, Backstein D, Davis A, Zukor DJ, Gross AE. Fresh osteochondral allografts for post-traumatic defects in the knee. A survivorship analysis. J Bone Joint Surg Br 1992; 74 (1) 105-110
    PubMedGoogle Scholar
  • 7 Teng MS, Yuen AS, Kim HT. Enhancing osteochondral allograft viability: effects of storage media composition. Clin Orthop Relat Res 2008; 466 (8) 1804-1809
    CrossrefPubMedGoogle Scholar
  • 8 Ball ST, Amiel D, Williams SK , et al. The effects of storage on fresh human osteochondral allografts. Clin Orthop Relat Res 2004; (418) 246-252
    PubMedGoogle Scholar
  • 9 Black J, Shadle CA, Parsons JR, Brighton CT. Articular cartilage preservation and storage. II. Mechanical indentation testing of viable, stored articular cartilage. Arthritis Rheum 1979; 22 (10) 1102-1108
    CrossrefPubMedGoogle Scholar
  • 10 Brighton CT, Shadle CA, Jimenez SA, Irwin JT, Lane JM, Lipton M. Articular cartilage preservation and storage. I. Application of tissue culture techniques to the storage of viable articular cartilage. Arthritis Rheum 1979; 22 (10) 1093-1101
    CrossrefPubMedGoogle Scholar
  • 11 Thomas VJ, Jimenez SA, Brighton CT, Brown N. Sequential changes in the mechanical properties of viable articular cartilage stored in vitro. J Orthop Res 1984; 2 (1) 55-60
    CrossrefPubMedGoogle Scholar
  • 12 Pennock AT, Wagner F, Robertson CM, Harwood FL, Bugbee WD, Amiel D. Prolonged storage of osteochondral allografts: does the addition of fetal bovine serum improve chondrocyte viability?. J Knee Surg 2006; 19 (4) 265-272
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 13 Linn MS, Chase DC, Healey RM, Harwood FL, Bugbee WD, Amiel D. Etanercept enhances preservation of osteochondral allograft viability. Am J Sports Med 2011; 39 (7) 1494-1499
    CrossrefPubMedGoogle Scholar
  • 14 Williams SK, Amiel D, Ball ST , et al. Prolonged storage effects on the articular cartilage of fresh human osteochondral allografts. J Bone Joint Surg Am 2003; 85-A (11) 2111-2120
    Google Scholar
  • 15 Amiel D, Harwood FL, Hoover JA, Meyers M. A histological and biochemical assessment of the cartilage matrix obtained from in vitro storage of osteochondral allografts. Connect Tissue Res 1989; 23 (1) 89-99
    CrossrefPubMedGoogle Scholar
  • 16 Bastian JD, Egli RJ, Ganz R, Hofstetter W, Leunig M. Chondrocytes within osteochondral grafts are more resistant than osteoblasts to tissue culture at 37°C. J Invest Surg 2011; 24 (1) 28-34
    CrossrefPubMedGoogle Scholar
  • 17 Williams JM, Virdi AS, Pylawka TK, Edwards III RB, Markel MD, Cole BJ. Prolonged-fresh preservation of intact whole canine femoral condyles for the potential use as osteochondral allografts. J Orthop Res 2005; 23 (4) 831-837
    CrossrefPubMedGoogle Scholar
  • 18 Pennock AT, Robertson CM, Wagner F, Harwood FL, Bugbee WD, Amiel D. Does subchondral bone affect the fate of osteochondral allografts during storage?. Am J Sports Med 2006; 34 (4) 586-591
    CrossrefPubMedGoogle Scholar
  • 19 McCarty WJ, Pallante AL, Rone RJ, Bugbee WD, Sah RL. The proteoglycan metabolism of articular cartilage in joint-scale culture. Tissue Eng Part A 2010; 16 (5) 1717-1727
    CrossrefPubMedGoogle Scholar
  • 20 Williams III RJ, Dreese JC, Chen CT. Chondrocyte survival and material properties of hypothermically stored cartilage: an evaluation of tissue used for osteochondral allograft transplantation. Am J Sports Med 2004; 32 (1) 132-139
    CrossrefPubMedGoogle Scholar
  • 21 Malinin T, Temple HT, Buck BE. Transplantation of osteochondral allografts after cold storage. J Bone Joint Surg Am 2006; 88 (4) 762-770
    CrossrefPubMedGoogle Scholar
  • 22 Allen RT, Robertson CM, Pennock AT , et al. Analysis of stored osteochondral allografts at the time of surgical implantation. Am J Sports Med 2005; 33 (10) 1479-1484
    CrossrefPubMedGoogle Scholar
  • 23 Csönge L, Bravo D, Newman-Gage H , et al. Banking of osteochondral allografts, Part II. Preservation of Chondrocyte Viability During Long-Term Storage. Cell Tissue Bank 2002; 3 (3) 161-168
    CrossrefPubMedGoogle Scholar
  • 24 Robertson CM, Allen RT, Pennock AT, Bugbee WD, Amiel D. Upregulation of apoptotic and matrix-related gene expression during fresh osteochondral allograft storage. Clin Orthop Relat Res 2006; 442: 260-266
    CrossrefPubMedGoogle Scholar
  • 25 Ohlendorf C, Tomford WW, Mankin HJ. Chondrocyte survival in cryopreserved osteochondral articular cartilage. J Orthop Res 1996; 14 (3) 413-416
    CrossrefPubMedGoogle Scholar
  • 26 Gole MD, Poulsen D, Marzo JM, Ko SH, Ziv I. Chondrocyte viability in press-fit cryopreserved osteochondral allografts. J Orthop Res 2004; 22 (4) 781-787
    CrossrefPubMedGoogle Scholar
  • 27 Jomha NM, Law GK, McGann LE. Storage of porcine articular cartilage at high subzero temperatures. Cell Tissue Bank 2006; 7 (1) 55-60
    CrossrefPubMedGoogle Scholar
  • 28 Acosta CA, Izal I, Ripalda P, Forriol F. Cell viability and protein composition in cryopreserved cartilage. Clin Orthop Relat Res 2007; 460: 234-239
    PubMedGoogle Scholar
  • 29 Judas F, Rosa S, Teixeira L, Lopes C, Ferreira Mendes A. Chondrocyte viability in fresh and frozen large human osteochondral allografts: effect of cryoprotective agents. Transplant Proc 2007; 39 (8) 2531-2534
    CrossrefPubMedGoogle Scholar
  • 30 Brockbank KG, Chen ZZ, Song YC. Vitrification of porcine articular cartilage. Cryobiology 2010; 60 (2) 217-221
    CrossrefPubMedGoogle Scholar
  • 31 Jomha NM, Weiss AD, Fraser Forbes J, Law GK, Elliott JA, McGann LE. Cryoprotectant agent toxicity in porcine articular chondrocytes. Cryobiology 2010; 61 (3) 297-302
    CrossrefPubMedGoogle Scholar
  • 32 Forriol F, Longo UG, Alvarez E , et al. Scanty integration of osteochondral allografts cryopreserved at low temperatures with dimethyl sulfoxide. Knee Surg Sports Traumatol Arthrosc 2011; 19 (7) 1184-1191
    CrossrefPubMedGoogle Scholar
  • 33 Chase D, Gortz S, Pallante A , et al. Histological and Biochemical Characterization of Fresh vs. Frozen Osteochondral Allografts after Six Months In Vivo in a Goat Model Paper presented at: Annual Meeting of the Orthopaedic Research Society 2011; Long Beach, CA
    PubMed
  • 34 Chase D, Görtz S, Pallante AL , et al. Histological and Biochemical Characterization of Fresh vs. Frozen Osteochondral Allografts after Six Months In Vivo in a Goat Model. Orthop Res Soc (ORS) Meeting 2011; Long Beach, CA (Poster 2098)
    PubMed
  • 35 Stoker A, Garrity J, Sims H, Cook J. Optimization of Osteochondral Allograft Preservation to Extend the Usable Life Span of Harvested Tissue. Paper presented at: Annual Meeting of the Orthopaedic Research Society 2011; Long Beach, CA
    PubMedGoogle Scholar
  • 36 Pallante AL, Bae WC, Chen AC, Görtz S, Bugbee WD, Sah RL. Chondrocyte viability is higher after prolonged storage at 37 degrees C than at 4 degrees C for osteochondral grafts. Am J Sports Med 2009; 37 (Suppl. 01) 24S-32S
    CrossrefPubMedGoogle Scholar
  • 37 Bian L, Stoker AM, Marberry KM, Ateshian GA, Cook JL, Hung CT. Effects of dexamethasone on the functional properties of cartilage explants during long-term culture. Am J Sports Med 2010; 38 (1) 78-85
    CrossrefPubMedGoogle Scholar
  • 38 López C, Ajenjo N, Muñoz-Alonso MJ, Farde P, León J, Gómez-Cimiano J. Determination of viability of human cartilage allografts by a rapid and quantitative method not requiring cartilage digestion. Cell Transplant 2008; 17 (7) 859-864
    CrossrefPubMedGoogle Scholar
  • 39 van der Valk J, Brunner D, De Smet K , et al. Optimization of chemically defined cell culture media—replacing fetal bovine serum in mammalian in vitro methods. Toxicol In Vitro 2010; 24 (4) 1053-1063
    CrossrefPubMedGoogle Scholar
  • 40 Friedlaender GE, Horowitz MC. Immune responses to osteochondral allografts: nature and significance. Orthopedics 1992; 15 (10) 1171-1175
    PubMedGoogle Scholar