Comparative evaluation of bioburden and sterility of indigenously prepared bone allograft with and without gentamicin

Abstract

During bone allograft processing, despite stringent donor screening and use of aseptic techniques, microbial invasion may occur due to the porous nature of the graft and cause potentially fatal infections. The aim of the present study was to prepare bone allograft with and without gentamicin and to compare bioburden and sterility in the obtained grafts to evaluate the role of antibiotic in enhancing graft safety. Fifty samples of demineralized freeze-dried bone allograft were prepared from suitable donors according to international standards. Randomly selected 25 samples were placed in 8 mg gentamicin/gram bone solution for 1 h. Packaging and sealing was done to ensure no microbial ingress during transportation. 40 samples were selected for bioburden testing. Remaining 10 were subjected to 25 kGy gamma radiation and tested for sterility. Microbiological evaluation revealed no evidence of colony forming units in all the samples of both the groups (Bioburden = 0). Post-radiation sterility testing also revealed no bacterial colony in the tested samples from both the groups. Favorable results validate the processing protocol while comparable results in both groups indicate no additive benefit of gentamicin addition. Nil bioburden may be used in further studies to determine a lower radiation dose to achieve adequate sterility and minimize the disadvantages of radiation like collagen cross-linking and decreased osteoinductive capacity.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

References

  1. AATB (2002) Standards for tissue banking. American Association of Tissue Banking, Maryland

    Google Scholar 

  2. Anagnostakos K, Wilmes P, Schmitt E, Kelm J (2009) Elution of gentamicin and vancomycin from polymethylmethacrylate beads and hip spacers in vivo. Acta Orthop 80:193–197

    Article  Google Scholar 

  3. Atal S et al (2016) Cost analysis of commonly used drugs under price control in India: assessing the effect of drug price control order on brand price variation. Int J Pharm Pharm Sci 8(4):315–321

    Google Scholar 

  4. Binte Atique F, Rahman Khalil MM (2014) The bacterial contamination of allogeneic bone and emergence of multidrug-resistant bacteria in tissue bank. Biomed Res Int 2014:430581

    Article  Google Scholar 

  5. Campana V, Milano G, Pagano E, Barba M, Cicione C, Salonna G, Lattanzi W, Logroscino G (2014) Bone substitutes in orthopaedic surgery: from basic science to clinical practice. J Mater Sci Mater Med 25:2445–2461

    CAS  Article  Google Scholar 

  6. Catto M (1965) A histological study of avascular necrosis of the femoral head after transcervical fracture. J Bone Joint Surg Br 47(4):749–776

    CAS  Article  Google Scholar 

  7. Chambers WB, Pallagrosi AU (1977) Gentamicin in the treatment of staphylococcal infections. J Int Med Res 5(6):442–449

    CAS  PubMed  Google Scholar 

  8. Coraça-Huber DC, Hausdorfer J, Fille M, Nogler M (2013) Effect of storage temperature on gentamicin release from antibiotic-coated bone chips. Cell Tissue Bank 14(3):395–400

    Article  Google Scholar 

  9. EAMST (2005) Standards. European Association of Musculo Skeletal Transplantation. http://www.eam-st.org/standards.htm. Accessed 17 May

  10. El Deeb M, Hosny M, Sharawy M (1989) Osteogenesis in composite grafts of allogenic demineralized bone powder and porous hydroxylapatite. J Oral Maxillofac Surg 47:50–56

    Article  Google Scholar 

  11. Fölsch C, Mittelmeier W, Bilderbeek U, Timmesfeld N, von Garrel T, Peter Matter H (2012) Effect of storage temperature on allograft bone. Transfus Med Hemother 39:36–40

    Article  Google Scholar 

  12. Gendler E (1986) Perforated demineralized bone matrix: a new form of osteoinductive material. J Biomed Mater Res 20:687–697

    CAS  Article  Google Scholar 

  13. Hilmy N, Febrida A, Basril A (2000) Validation of radiation sterilization dose for lyophilized amnion and bone grafts. Cell Tissue Bank 1(2):143–148

    CAS  Article  Google Scholar 

  14. IAEA (2002) International standards on tissue banking – first meeting of the IAEA technical advisory committee. In: Committee ITA (ed) The IAE Aprogramme in radiation and tissue banking. IAEA, Boston, USA, pp 44

  15. Isefuku S, Joyner CJ, Simpson AH (2003) Gentamicin may have an adverse effect on osteogenesis. J Orthop Trauma 17(3):212–216

    Article  Google Scholar 

  16. Kamra P, Lamba AK, Faraz F, Thakur A (2017) Effect of antibiotic impregnation time on the release of gentamicin from cryopreserved allograft bone chips—an in vitro study. Dissertation, University of Delhi

  17. Ketonis C, Hickok N, Parvizi J (2011) Antimicrobial implants and bone allografts: new uses for old antibiotics. Science against microbial pathogens: communicating current research and technological advances. In: Mendez-Vilas A (ed) Science against microbial pathogens. Badajoz, Formatex Research Center, pp 469–482

    Google Scholar 

  18. Liu JW, Chao LH, Su LH, Wang JW, Wang CJ (2002) Experience with a bone bank operation and allograft bone infection in recipients at a medical centre in southern Taiwan. J Host Infect 50:293–297

    CAS  Article  Google Scholar 

  19. Lord CF, Gebhardt MC, Tomford WW, Mankin HJ (1988) Infection in bone allografts: incidence, nature, and treatment. J Bone Joint Surg Am 70(3):369–376

    CAS  Article  Google Scholar 

  20. Mellonig JT, Prewett AB, Moyer MP (1992) HIV inactivation in a bone allograft. J Periodontol 63(12):979–983

    CAS  Article  Google Scholar 

  21. Nather A, Yusof N, Hilmy N (2010) Allograft procurement, processing and transplantation. World Scientific Publishing Company, Singapore

    Google Scholar 

  22. Nguyen H, Morgan DA, Sly LI, Benkovich M, Cull S, Forwood MR (2008) Validation of 15 kGy as a radiation sterilisation dose for bone allografts manufactured at the Queensland Bone Bank: application of the VDmax 15 method. Cell Tissue Bank 9(2):139–147

    CAS  Article  Google Scholar 

  23. Pearson K, Dock N, Brubaker S (2008) Standards for tissue banking, 12th edn. American Association of Tissue Banks, McLean, Virginia

    Google Scholar 

  24. Pietrzak WS, Ali SN, Chitturi D, Jacob M, Woodell-May JE (2011) BMP depletion occurs during prolonged acid demineralization of bone: characterization and implications for graft preparation. Cell Tissue Bank 12(2):81–88

    CAS  Article  Google Scholar 

  25. Richards F, Cox C (1971) Gentamicin treatment of staphylococcal infections. JAMA 215(8):1297–1300

    CAS  Article  Google Scholar 

  26. Ronholdt CJ, Bogdansky S, Baker T (2005) Establishing an Appropriate terminal Sterilisation Dose Based upon Post-Processing Bioburden Levels on Allograft Tissue. In: Kennedy FJ, Phillips OG, Williams AP (eds) Sterilisation of tissues using ionising radiation. CRC Press, Cambridge, pp 311–318

    Google Scholar 

  27. Strong DM (2000) The US Navy Tissue Bank: 50 years on the cutting edge. Cell Tissue Bank 1(1):9–16

    CAS  Article  Google Scholar 

  28. Sutherland AG, Raafat A, Yates P, Hutchison JD (1997) Infection associated with the use of allograft bone from the north east Scotland Bone Bank. J Hosp Infect 35:215–222

    CAS  Article  Google Scholar 

  29. Thorén K, Aspenberg P, Thorngren KG (1993) Lipid extraction decreases the specific immunologic response to bone allografts in rabbits. Acta Orthop Scand 64(1):44–46

    Article  Google Scholar 

  30. Tian J, Zhou Z, Lian Y, Pei Yang Y, Shen B (2009) Effects of three defatting ways on porcine bone. J Clin Rehabilit Tissue Eng Res 13(16):3133–3136

    CAS  Google Scholar 

  31. van de Pol GJ, Sturm PD, van Loon CJ, Verhagen C, Schreurs BW (2007) Microbiological cultures of allografts of the femoral head just before transplantation. J Bone Joint Surg Br 89(9):1225–1228

    PubMed  Google Scholar 

  32. Witsø E, Persen L, Benum P, Bergh K (2002) Release of netilmicin and vancomycin from cancellous bone. Acta Orthop Scand 73(2):199–205

    Article  Google Scholar 

  33. Young SE, Wilkins RM (1995) Medical/social history questionnaires; validating the process. In: Proceedings of 19th annual meeting. American Association of Tissue Banks. Atlanta, GA

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Kanika Makker.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with animals performed by any of the authors. All procedures performed on human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments.

Funding

This research did not receive any grant from funding agencies in the public, commercial, or not for-profit sectors.

Informed consent

Informed consent was obtained from all human tissue donors.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Makker, K., Lamba, A.K., Faraz, F. et al. Comparative evaluation of bioburden and sterility of indigenously prepared bone allograft with and without gentamicin. Cell Tissue Bank 20, 243–253 (2019). https://doi.org/10.1007/s10561-019-09763-w

Download citation

Keywords

  • Bone allograft
  • Bioburden
  • Gentamicin
  • Gamma radiation