Cell and Tissue Banking

, Volume 19, Issue 4, pp 603–612 | Cite as

Effects of supercritical fluid CO2 and 25 kGy gamma irradiation on the initial mechanical properties and histological appearance of tendon allograft

  • Mazyar Irani
  • Vedran LovricEmail author
  • William R. Walsh


Tendon allografts, when autograft options are limited or when obtaining an autograft is not aligned with the patients’ best interest, play an important role in tendon and ligament reconstruction. To minimize the risk of infectious disease transmission tissue banks perform screening tests and the allografts cleaned are sterilized. The current study examines and compares the initial mechanical properties and histological appearance of supercritical CO2 (SCCO2)-treated and gamma-irradiated porcine extensor tendons. Thirty intact porcine forelimb extensor tendons randomized equally into three groups: control group, gamma-irradiation group, and SCCO2-treated group. Once treated, histological assessment and histomorphologic measurements were made on the histological sections obtained from each tendon while stiffness and ultimate failure loads were evaluated from tensile testing. Histological evaluation of gamma-irradiated tendons showed significant disruption to the hierarchical morphology of the fascicle bundles, which was not evident in SCCO2-treated specimens. Histomorphologic measurements showed a significant increase for measured dead space (void) between tendon fibrils of the gamma-irradiated group comparing to both control and SCCO2 treated groups (p < 0.01). There was a significant reduction in the ultimate failure load for tendons treated by gamma-irradiation compared to the control group (p < 0.05). No statistically significant difference was detected between control and SCCO2-treated tendons in the ultimate failure load. Stiffness values were not significantly different between three-study groups. This study suggests that while gamma-irradiation has a deleterious effect on mechanical properties of tendon tissue, SCCO2 does not alter the biomechanical properties and the histological structure of porcine extensor tendons.


Supercritical CO2 (SCCO2Gamma-irradiation Stiffness Ultimate failure loads Soft-tissue processing Tendon allograft 


Compliance with ethical standards

Conflict of interest

The authors of this study clarify that they have not accepted any funds for this study and had no conflict of interest related to this study to declare.


  1. Aydin A, Tuncer S, Eeer M, Bilgic B (2004) Tumoural calcinosis infiltrating the biceps brachii tendon: excision and reconstruction with allograft. J Hand Surg [Br] 29-B:170–172CrossRefGoogle Scholar
  2. Baldini T, Caperton Hawkins M, McCarty E (2014) Effect of novel sterilization method of biomechanical properties of soft tissue allografts. Knee Surg Sports Traumatol Arthroscop. CrossRefGoogle Scholar
  3. Beer FP, Johnston R (2006) Mechanics of materials. McGraw-Hill, New York. ISBN 978-0-07-352938-7Google Scholar
  4. Bhatia S, Bell R, Frank RM, Rodeo SA, Bach BR Jr, Cole BJ, Chubinskaya S, Wang VM, Verma NN (2012) Bony incorporation of soft tissue anterior cruciate ligament grafts in an animal model: autograft versus allograft with low-dose gamma irradiation. Am J Sports Med 40(8):1789–1798CrossRefPubMedGoogle Scholar
  5. Bui D, Lovric V, Oliver R, Bertollo N, Broe D, Walsh WR (2015) Meniscal allograft sterilisation: effect on biomechanical and histological properties. Cell Tissue Bank 16:467–475. CrossRefPubMedGoogle Scholar
  6. Canale ST, Azar FM, Beaty HD (2016) Campbell’s operative orthopaedics. Elsevier, Amsterdam. ISBN 978-0323072434Google Scholar
  7. Celli A, Arash A, Adams RA, Morrey BF (2005) Triceps insufficiency following total elbow arthroplasty. J Bone Joint Surg [Am] 87-A:1957–1964CrossRefGoogle Scholar
  8. Cheung DT, Perelman N, Tong D, Nimni ME (1990) The effect of gamma irradiation on collagen molecules, isolated alpha-chains and cross-linked native fibers. J Biomed Mater Res 24:581–589CrossRefPubMedGoogle Scholar
  9. Christensen TW, Burns D, White A, Ganem B, Eisenhunt A (2004) Sterilization methods and apparatus which employ additive-containing supercritical carbon dioxide sterilant. US 7108832 B2, 2006Google Scholar
  10. Conrad BP, Rappe M, Horodyski M, Farmer KW, Indelicato PA (2013) The effect of sterilization on mechanical properties of soft tissue allografts. Cell Tissue Bank 14(3):359–366CrossRefPubMedGoogle Scholar
  11. Edwards JH, Herbert A, Jones GL, Manfield IW, Fisher J, Ingham E (2016) The effects of irradiation on the biological and biomechanical properties of an acellular porcine superflexor tendon graft for cruciate ligament repair. J Biomed Mater Res Part B Appl Biomater. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Fages J, Poirier B, Barbier Y, Frayssinet P, Joffret ML, Majewski W, Bonel G, Larzul D (1996) Viral inactivation of human bone tissue using supercritical fluid extraction. ASAIO J 44(4):289–293Google Scholar
  13. Fideler BM, Vangsness CT Jr, Moore T, Li Z, Rasheed S (1994) Effects of gamma irradiation on the human immunodeficiency virus. A study in frozen human bone-patellar ligament-bone grafts obtained from infected cadavera. J Bone Joint Surg Am 76(7):1032–1035CrossRefGoogle Scholar
  14. Furukawa S, Watanabe T, Koyama T, Hirata J, Narisawa N, Ogihara H, Yamasaki M (2009) Inactivation of food poioning bacteria and Geobacillus stearothermophilus spores by high pressure carbon dioxide treatment. Food Control 20:53–58. CrossRefGoogle Scholar
  15. Gouk SS, Lim TM, Teoh SH, Sun WQ (2008) Alterations of human acellular tissue matrix by gamma irradiation: histology, biomechanical property, stability, in vitro cell repopulation and remodeling. J Biomed Mat Res Part B Appl Biomater 84B:205–217CrossRefGoogle Scholar
  16. Greis PE, Koch BS, Adams B (2012) Tibialis anterior or posterior allograft anterior cruciate ligament reconstruction versus hamstring autograft reconstruction: an economic analysis in a hospital-based outpatient setting. Arthroscopy 28(11):1695–1701CrossRefPubMedGoogle Scholar
  17. Guo L, Yang L, Duan XJ, He R, Chen GX, Wang FY, Zhang Y (2012) Anterior cruciate ligament reconstruction with bonepatellar tendon-bone graft: comparison of autograft, fresh-frozen allograft, and gamma-irradiated allograft. Arthroscopy 28(2):211–217CrossRefPubMedGoogle Scholar
  18. Gupta HS, Seto J, Krauss S, Boesecke P, Screen HRC (2010) In situ multi-level analysis of viscoelastic deformation mechanisms in tendon collagen. J Struct Biol 169(2):183–191CrossRefPubMedGoogle Scholar
  19. Hernigou P, Gras G, Marinello G, Dormont D (2000) Influence of irradiation on the risk of transmission of HIV in bone grafts obtained from appropriately screened donors and followed by radiation sterilization. Cell Tissue Bank 1(4):279–289CrossRefPubMedGoogle Scholar
  20. Hoburg AT, Keshlaf S, Schmidt T, Smith M, Gohs U, Perka C, Pruss A, Scheffler S (2010) Effect of electron beam irradiation on biomechanical properties of patellar tendon allografts in anterior cruciate ligament reconstruction. Am J Sports Med 38(6):1134–1140CrossRefPubMedGoogle Scholar
  21. Huang Q, Ingham E, Rooney P, Kearney JN (2013) Production of a sterilised decellularised tendon allograft for clinical use. Cell Tissue Bank 14(4):645–654CrossRefGoogle Scholar
  22. Hulmes DJS et al (2002) Building collagen molecules, fibrils, and suprafibrillar structures. J Struct Biol 137(1–2):2–10. CrossRefPubMedGoogle Scholar
  23. Inoue N, Bessho M, Furuta M, Kojima T, Okuda S, Hara M (2006) A novel collagen hydrogel cross-linked by gamma-ray irradiation in acidic pH conditions. J Biomater Sci Polym Ed 17:837–858CrossRefPubMedGoogle Scholar
  24. Joseph TA, Defranco MJ, Weiker GG (2003) Delayed repair of a pectoralis major tendon rupture with allograft: a case report. J Shoulder Elbow Surg 12:101–104CrossRefPubMedGoogle Scholar
  25. Keller PF, Verin V, Ziegler T, Mermillod B, Popowski Y, Delafontaine P (2001) Gamma-irradiation markedly inhibits the hydrated collagen gel contradiction by arterial smooth muscle cells. J Invest Med 49:258–264CrossRefGoogle Scholar
  26. Liu BC, Harrell R, Davis RH, Dresden MH, Spira M (1989) The effect of gamma irradiation on injectable human amnion collagen. J Biomed Mater Res 23:833–844CrossRefPubMedGoogle Scholar
  27. Lovric V, Kanazawa T, Nakamura Y, Oliver R, Yu Y, Walsh WR (2011) Effects of gaps induced into the ACL tendon graft on tendon-bone healing in a rodent ACL reconstruction model. Muscle Tendon Ligament J 1(3):91–99Google Scholar
  28. McAllister DR, Joyce MJ, Mann BJ, Vangsness CT Jr (2007) Allograft update: the current status of tissue regulation, procurement, processing, and sterilization. Am J Sports Med 35:2148–2158CrossRefGoogle Scholar
  29. McGilvray KC, Santoni BG, Turner AS, Bogdansky S, Wheeler DL, Puttlitz CM (2011) Effects of 60Co gamma radiation dose on initial structural biomechanical properties of ovine bone-patellar tendon-bone allografts. Cell Tissue Bank 12:89–98CrossRefPubMedGoogle Scholar
  30. Melo Silva J, Rigo AA, Dalmolin IA, Debien I, Cansian RL, Oliveira JV, Mazutti MA (2013) Effect of pressure, depressurization rate and pressure cycling on the inactivation of Escherichia coli by supercritical carbon dioxide. Food Control 29:76–81. CrossRefGoogle Scholar
  31. Meyerhof WE et al (1967) Elements of nuclear physics. McGraw-Hill, New York. ISBN 978-0070417458Google Scholar
  32. Nakata K, Shino K, Horibe S et al (2000) Reconstruction of the lateral ligaments of the ankle using solvent-dried and gamma-irradiated allogeneic fascia lata. J Bone Joint Surg [Br] 82-B:579–582CrossRefGoogle Scholar
  33. Ota T, Taketani S, Iwai S, Miyagawa S, Furuta M, Hara M et al (2007) Novel method of decellularization of porcine valves using polyethylene glycol and gamma irradiation. Ann Thorac Surg 83:1501–1507CrossRefPubMedGoogle Scholar
  34. Padrela L, Rodrigues MA, Velaga SP, Matos HA, Azevedo EG (2009) Formation of indomethacin–saccharin cocrystals using supercritical fluid technology. Eur J Pharm Sci 38:9–17. CrossRefPubMedGoogle Scholar
  35. Perrut M et al (2012) Sterilization and virus inactivation by supercritical fluids (a review). J Supercrit Fluids 66:359–371. CrossRefGoogle Scholar
  36. Puxkandl R, Zizak I, Paris O, Keckes J, Tesch W, Bernstorff S, Purslow P, Fratzl P (2002) Viscoelastic properties of collagen: synchrotron radiation investigations and structural model. Philos Trans R Soc 357(1418):191–197. CrossRefGoogle Scholar
  37. Robertson A, Nutton RW, Keating JF (2006) Current trends in the use of tendon allografts in orthopaedic surgery. J Bone Joint Surg [Br] 88-B:988–992CrossRefGoogle Scholar
  38. Russell NA, Pelletier MH, Bruce WJ, Walsh WR (2012a) The effect of gamma irradiation on the anisotropy of bovine cortical bone. Med Eng Phys 34:1117–1122. CrossRefPubMedGoogle Scholar
  39. Russell NA, Rives A, Pelletier MH, Bruce WJ, Walsh WR (2012b) The effect of sterilization on the mechanical properties of intact rabbit humeri in three-point bending, four-point bending and torsion. Cell Tissue Bank. CrossRefPubMedGoogle Scholar
  40. Sanchez-Sotelo J, Morrey BF (2002) Surgical techniques for reconstruction of chronic insufficiency of the triceps: rotation flap using anconeus and tendo achillis allograft. J Bone Joint Surg [Br] 84-B:1116–1120CrossRefGoogle Scholar
  41. Sanchez-Sotelo J, Morrey BF, Adams RA, O’Driscoll SW (2002) Reconstruction of chronic ruptures of the distal biceps tendon with use of an achilles tendon allograft. J Bone Joint Surg [Am] 84-A:999–1005CrossRefGoogle Scholar
  42. Schimizzi A, Wedemeyer M, Odell T, Thomas W, Mahar AT, Pedowitz R (2007) Effects of a novel sterilization process on soft tissue mechanical properties for anterior cruciate ligament allografts. Am J Sports Med 35(4):612–616CrossRefPubMedGoogle Scholar
  43. Schmidt T, Hoburg A, Broziat C, Smith MD, Gohs U, Pruss A, Scheffler S (2012) Sterilization with electron beam irradiation influences the biomechanical properties and the early remodeling of tendon allografts for reconstruction of the anterior cruciate ligament (ACL). Cell Tissue Bank 13(3):387–400CrossRefGoogle Scholar
  44. Screen H, Lee DA, Bader DL, Shelton JC (2004) An investigation into the effects of the hierarchical structure of tendon fascicles on micromechanical properties. J Eng Med 218:109–119CrossRefGoogle Scholar
  45. Seto AU, Culp BM, Gatt CJ Jr, Dunn M (2013) Radioprotection provides functional mechanics but delays healing of irradiated tendon allografts after ACL reconstruction in sheep. Cell Tissue Bank 14(4):655–665CrossRefPubMedGoogle Scholar
  46. Su EP, Healey JH (2003) Salvage reconstruction for lateral ankle instability using a tendon allograft. Clin Orthop 415:232–238CrossRefGoogle Scholar
  47. Sun K, Zhang J, Wang Y, Xia C, Zhang C, Yu T, Tian S (2011) Arthroscopic anterior cruciate ligament reconstruction with at least 2.5 years’ follow-up comparing hamstring tendon autograft and irradiated allograft. Arthroscopy 27(9):1195–1202CrossRefPubMedGoogle Scholar
  48. TGA (2011) Regulatory life cycle for biologicals that are included on the Australian Register of Therapeutic Goods, vol 1, 1st edn. Australian Government, CanberraGoogle Scholar
  49. White A, Burns D, Christensen TW (2006) Effective terminal sterilization using supercritical carbon dioxide. J Biotechnol 123(4):504–515CrossRefPubMedGoogle Scholar
  50. Yanke AB, Bell R, Lee A, Kang RW, Mather RC III, Shewman EF, Wang VM, Bach BR Jr (2013) The biomechanical effects of 1.0 to 1.2 Mrad of gamma irradiation on human bone-patellar tendon-bone allografts. Am J Sports Med 41(4):835–840CrossRefPubMedGoogle Scholar
  51. Zhang GEY, Chervoneva I, Robinson PS, Beason DP, Carine ET, Soslowsky LJ, Iozzo RV, Birk DE (2006a) Decorin regulates assembly of collagen fibrils and acquisition of biomechanical properties during tendon development. J Cell Biochem 98(6):1436–1449. CrossRefPubMedGoogle Scholar
  52. Zhang J, Davis TA, Matthews MA, Drews MJ, LaBerge M, An YH (2006b) Sterilization using high-pressure carbon dioxide. J Supercrit Fluids 38:354–372. CrossRefGoogle Scholar
  53. Zhou M, Zhang N, Liu X, Li Y, Zhang Y, Wang X, Li B (2014) Tendon allograft sterilized by peracetic acid/ethanol combined with gamma irradiation. J Orthop Sci. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Mazyar Irani
    • 1
  • Vedran Lovric
    • 1
    Email author
  • William R. Walsh
    • 1
  1. 1.Surgical and Orthopaedic Research Laboratories Prince of Wales Clinical School, Prince of Wales HospitalUniversity of New South WalesSydneyAustralia

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