Abstract
Bone allografts are clinically used in a variety of surgical procedures, and tissue banks are responsible for harvesting, processing, quality testing, storing, and delivering these materials for transplantation. In tissue banks, the bone is processed for the removal of all organic content, remaining only the tissue structure (scaffold). However, several studies have shown that even after using different processing methods, viable cells, functional proteins, and DNA may still persist in the tissue, which constitute the main causes of graft rejection. Therefore, the objective of this study was to establish techniques and biological parameters for quality validation of allografts. To this end, we propose the use of 3 combined methods such as microscopy, histology, and molecular biology techniques to evaluate the quality of allografts harvested and processed by the Brazilian National Institute of Traumatology and Orthopedics (INTO) tissue bank according to the donation criteria of the Brazilian National Health Surveillance Agency and the Brazilian National Transplant System. Bone fragments from different processing stages showed no viable cells on histology, an intact extracellular matrix on scanning electron microscopy, and gradual reduction in DNA amount. Different techniques were used to demonstrate the quality of allografts produced by the INTO tissue bank and to establish biological parameters for ensuring the safety and quality of these products. Future studies need to be undertaken to assess and validate the efficacy of the decellularization process in larger bone grafts with diverse architectural configurations.
Similar content being viewed by others
Data availability
All relevant data are within the paper.
References
Badylak SF, Gilbert TW. (2008) Immune response to biologic scaffold materials. Semin Immunol. 20(2):109–16. https://doi.org/10.1016/j.smim.2007.11.003
Chen G, Lv Y (2017) Decellularized bone matrix scaffold for bone regeneration. In: Turksen K (ed) Decellularized scaffolds and organogenesis. Methods in molecular biology. Springer, New York, pp 239–254
Civril F, Deimling T, de Oliveira Mann CC, Ablasser A, Moldt M, Witte G, Hornung V, Hopfner KP (2013) Structural mechanism of cytosolic DNA sensing by cGAS. Nature 498:332–337. https://doi.org/10.1038/nature12305
Coutinho LF, Amaral JBD, Santos EBD, Martinez EF, Montalli VAM, Junqueira JLC, Araujo VC, Napimoga MH (2017) Presence of cells in fresh-frozen allogeneic bone grafts from different tissue banks. Braz Dent J 28:152–157. https://doi.org/10.1590/0103-6440201701206
Crapo PM, Gilbert TW, Badylak SF (2011) An overview of tissue and whole organ decellularization processes. Biomaterials 32:3233–3243. https://doi.org/10.1016/j.biomaterials.2011.01.057
Curtis EM, van der Velde R, Moon RJ, van den Bergh JP, Geusens P, de Vries F, van Staa TP, Cooper C, Harvey NC (2016) Epidemiology of fractures in the United Kingdom 1988–2012: Variation with age, sex, geography, ethnicity and socioeconomic status. Bone 87:19–26. https://doi.org/10.1016/j.bone.2016.03.006
da Rocha LR, Dias RB, Fernandes MBC, Prinz R, Eirado TP, Costa IS, Monteiro MJ, da Silva CER, Dos Santos CT, Fogagnolo F (2023) A new option for bone regeneration: a rapid methodology for cellularization of allograft with human bone marrow stromal cells with in vivo bone-forming potential. Injury 54(Suppl 6):110777. https://doi.org/10.1016/j.injury.2023.05.008
Damien CJ, Parsons JR (1991) Bone graft and bone graft substitutes: a review of current technology and applications. J Appl Biomater 2:187–208. https://doi.org/10.1002/jab.770020307
De Lacerda PE, Pelegrine AA, Teixeira ML, Montalli VA, Rodrigues H, Napimoga MH (2016) Homologous transplantation with fresh frozen bone for dental implant placement can induce HLA sensitization: a preliminary study. Cell Tissue Bank 17:465–472. https://doi.org/10.1007/s10561-016-9562-9
Fretwurst T, Spanou A, Nelson K, Wein M, Steinberg T, Stricker A (2014) Comparison of four different allogeneic bone grafts for alveolar ridge reconstruction: a preliminary histologic and biochemical analysis. Oral Surg Oral Med Oral Pathol Oral Radiol 118:424–431. https://doi.org/10.1016/j.oooo.2014.05.020
Gardin C, Ricci S, Ferroni L, Guazzo R, Sbricoli L, De Benedictis G, Finotti L, Isola M, Bressan E, Zavan B (2015) Decellularization and delipidation protocols of bovine bone and pericardium for bone grafting and guided bone regeneration procedures. PLoS ONE 10:e0132344. https://doi.org/10.1371/journal.pone.0132344
Hashimoto Y, Funamoto S, Kimura T, Nam K, Fujisato T, Kishida A (2011) The effect of decellularized bone/bone marrow produced by high-hydrostatic pressurization on the osteogenic differentiation of mesenchymal stem cells. Biomaterials 32:7060–7067. https://doi.org/10.1016/j.biomaterials.2011.06.008
Hernigou P, Poignard A, Beaujean F, Rouard H (2005) Percutaneous autologous bone-marrow grafting for nonunions. Influence of the number and concentration of progenitor cells. J Bone Joint Surg Am 87:1430–1437. https://doi.org/10.2106/JBJS.D.02215
Keane TJ, Swinehart IT, Badylak SF (2015) Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance. Methods 84:25–34. https://doi.org/10.1016/j.ymeth.2015.03.005
Kim YS, Majid M, Melchiorri AJ, Mikos AG (2019) Applications of decellularized extracellular matrix in bone and cartilage tissue engineering. Bioeng Transl Med 4:83–95. https://doi.org/10.1002/btm2.10110
Lee DJ, Diachina S, Lee YT, Zhao L, Zou R, Tang N, Han H, Chen X, Ko CC (2016) Decellularized bone matrix grafts for calvaria regeneration. J Tissue Eng 7:2041731416680306. https://doi.org/10.1177/2041731416680306
Miranda C, Leonel L, Canada RR, Maria DA, Miglino MA, Del Sol M, Lobo SE (2021) Effects of chemical and physical methods on decellularization of murine skeletal muscles. An Acad Bras Cienc 93:e20190942. https://doi.org/10.1590/0001-3765202120190942
Rasch A, Naujokat H, Wang F, Seekamp A, Fuchs S, Kluter T (2019) Evaluation of bone allograft processing methods: impact on decellularization efficacy, biocompatibility and mesenchymal stem cell functionality. PLoS ONE 14:e0218404. https://doi.org/10.1371/journal.pone.0218404
Sohn HS, Oh JK (2019) Review of bone graft and bone substitutes with an emphasis on fracture surgeries. Biomater Res 23:9. https://doi.org/10.1186/s40824-019-0157-y
Urciuolo A, De Coppi P (2018) Decellularized tissue for muscle regeneration. Int J Mol Sci 19:2392. https://doi.org/10.3390/ijms19082392
Varettas K, Taylor P (2011) Bioburden assessment of banked bone used for allografts. Cell Tissue Bank 12:37–43. https://doi.org/10.1007/s10561-009-9154-z
Wainwright JM, Czajka CA, Patel UB, Freytes DO, Tobita K, Gilbert TW, Badylak SF (2010) Preparation of cardiac extracellular matrix from an intact porcine heart. Tissue Eng Part C Methods 16:525–532. https://doi.org/10.1089/ten.TEC.2009.0392
Wiese A, Pape HC (2010) Bone defects caused by high-energy injuries, bone loss, infected nonunions, and nonunions. Orthop Clin North Am 41:1–4. https://doi.org/10.1016/j.ocl.2009.07.003
Xu K, Kuntz LA, Foehr P, Kuempel K, Wagner A, Tuebel J, Deimling CV, Burgkart RH (2017) Efficient decellularization for tissue engineering of the tendon-bone interface with preservation of biomechanics. PLoS ONE 12:e0171577. https://doi.org/10.1371/journal.pone.0171577
Funding
The study was partially supported by the AO Trauma Latin America Research Office.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors have no conflicts of interest to disclose.
Human and animal participants
The research project was approved by the Institutional Research Ethics Committee (CAAE protocol no. 21778619.9.0000.5273). Study samples were harvested by the INTO tissue bank and are in accordance with the criteria for allograft donation of the Brazilian National Health Surveillance Agency (RDC 220/2006 ANVISA) and the Brazilian National Transplant System.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Prinz, R.A.D., da Rocha, L.R., Eirado, T.P. et al. Biological parameters for quality evaluation of allografts from the Brazilian National Institute of Traumatology and Orthopedics tissue bank. Cell Tissue Bank (2024). https://doi.org/10.1007/s10561-024-10125-4
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10561-024-10125-4