Abstract
Demineralized bone matrix (DBM) has been regarded as an ideal bone substitute as a native carrier of bone morphogenetic proteins (BMPs) and other growth factors. However, the osteoinductive properties diverse in different DBM products. We speculate that the harvest origin further contributing to variability of BMPs contents in DBM products besides the process technology. In the study, the cortical bone of femur, tibia, humerus, and ulna from a signal donor were prepared and followed demineralizd into DBM products. Proteins in bone martix were extracted using guanidine-HCl and collagenase, respectively, and BMP-2 content was detected by sandwich enzyme-linked immunosorbent assay (ELISA). Variability of BMP-2 content was found in 4 different DBM products. By guanidine-HCl extraction, the average concentration in DBMs harvested from ulna, humerus, tibia, and femur were 0.613 ± 0.053, 0.848 ± 0.051, 3.293 ± 0.268, and 21.763 ± 0.344, respectively (p < 0.05), while using collagenase, the levels were 0.089 ± 0.004, 0.097 ± 0.004, 0.330 ± 0.012, and 1.562 ± 0.008, respectively (p < 0.05). In general, the content of BMP-2 in long bones of Lower limb was higher than that in long bones of upper limb, and GuHCl had remarkably superior extracted efficiency for BMP-2 compared to collagenase. The results suggest that the origin of cortical bones harvested to fabricate DBM products contribute to the variability of native BMP-2 content, while the protein extracted method only changes the measured values of BMP-2.
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References
Alaee F, Hong SH, Dukas AG, Pensak MJ, Rowe DW, Lieberman JR (2014) Evaluation of osteogenic cell differentiation in response to bone morphogenetic protein or demineralized bone matrix in a critical sized defect model using GFP reporter mice. J Orthop Res 32:1120–1128. https://doi.org/10.1002/jor.22657
Bae HW, Zhao L, Kanim LE, Wong P, Delamarter RB, Dawson EG (2006) Intervariability and intravariability of bone morphogenetic proteins in commercially available demineralized bone matrix products. Spine (Phila Pa 1976) 31:1299–1308. https://doi.org/10.1097/01.brs.0000218581.92992.b7
Bae H, Zhao L, Zhu D, Kanim LE, Wang JC, Delamarter RB (2010) Variability across ten production lots of a single demineralized bone matrix product. J Bone Jt Surg Am 92:427–435. https://doi.org/10.2106/JBJS.H.01400
Bae HW, Zhao L, Kanim LE, Wong P, Marshall D, Delamarter RB (2013) Bone marrow enhances the performance of rhBMP-2 in spinal fusion: a rodent model. J Bone Jt Surg Am 95:338–347. https://doi.org/10.2106/JBJS.K.01118
Bal Z, Korkusuz F, Ishiguro H, Okada R, Kushioka J, Chijimatsu R et al (2021) A novel nano-hydroxyapatite/synthetic polymer/bone morphogenetic protein-2 composite for efficient bone regeneration. Spine J 21:865–873. https://doi.org/10.1016/j.spinee.2021.01.019
Bhamb N, Kanim LEA, Drapeau S, Mohan S, Vasquez E, Shimko D et al (2019) Comparative efficacy of commonly available human bone graft substitutes as tested for posterolateral fusion in an athymic rat model. Int J Spine Surg 13:437–458
Campana V, Milano G, Pagano E, Barba M, Cicione C, Salonna G et al (2014) Bone substitutes in orthopaedic surgery: from basic science to clinical practice. J Mater Sci Mater Med 25:2445–2461. https://doi.org/10.1007/s10856-014-5240-2
Dinopoulos HT, Giannoudis PV (2006) Safety and efficacy of use of demineralised bone matrix in orthopaedic and trauma surgery. Expert Opin Drug Saf 5:847–866. https://doi.org/10.1517/14740338.5.6.847
Drosos GI, Kazakos KI, Kouzoumpasis P, Verettas DA (2007) Safety and efficacy of commercially available demineralised bone matrix preparations: a critical review of clinical studies. Injury 38(Suppl 4):13–21. https://doi.org/10.1016/s0020-1383(08)70005-6
Drosos GI, Touzopoulos P, Ververidis A, Tilkeridis K, Kazakos K (2015) Use of demineralized bone matrix in the extremities. World J Orthop 6:269–277. https://doi.org/10.5312/wjo.v6.i2.269
Ellis SL, Grassinger J, Jones A, Borg J, Camenisch T, Haylock D et al (2011) The relationship between bone, hemopoietic stem cells, and vasculature. Blood 118:1516–1524. https://doi.org/10.1182/blood-2010-08-303800
Greenwald AS, Boden SD, Goldberg VM, Khan Y, Laurencin CT, Rosier RN (2001) Bone-graft substitutes: facts, fictions, and applications. J Bone Jt Surg Am 83:98–103. https://doi.org/10.2106/00004623-200100022-00007
Han B, Yang Z, Nimni M (2005) Effects of moisture and temperature on the osteoinductivity of demineralized bone matrix. J Orthop Res 23:855–861. https://doi.org/10.1016/j.orthres.2004.11.007
Honsawek S, Powers RM, Wolfinbarger L (2005) Extractable bone morphogenetic protein and correlation with induced new bone formation in an in vivo assay in the athymic mouse model. Cell Tissue Bank 6:13–23. https://doi.org/10.1007/s10561-005-1445-4
Huber E, Pobloth AM, Bormann N, Kolarczik N, Schmidt-Bleek K, Schell H et al (2017) Demineralized bone matrix as a carrier for bone morphogenetic protein-2: burst release combined with long-term binding and osteoinductive activity evaluated in vitro and in vivo. Tissue Eng Part 23:1321–1330. https://doi.org/10.1089/ten.TEA.2017.0005
Katz JM, Nataraj C, Jaw R, Deigl E, Bursac P (2009) Demineralized bone matrix as an osteoinductive biomaterial and in vitro predictors of its biological potential. J Biomed Mater Res B Appl Biomater 89:127–134. https://doi.org/10.1002/jbm.b.31195
Ku JK, Kim IH, Um IW, Kim BH, Yun PY (2022) Effect of gamma irradiation on the osteoinductivity of demineralized dentin matrix for allografts: a preliminary study. J Funct Biomater 13:14. https://doi.org/10.3390/jfb13010014
Loeffler BJ, Kellam JF, Sims SH, Bosse MJ (2012) Prospective observational study of donor-site morbidity following anterior iliac crest bone-grafting in orthopaedic trauma reconstruction patients. J Bone Joint Surg Am 94:1649–1654. https://doi.org/10.2106/JBJS.K.00961
Murray SS, Brochmann EJ, Harker JO, King E, Lollis RJ, Khaliq SA (2007) A statistical model to allow the phasing out of the animal testing of demineralised bone matrix products. Altern Lab Anim 35:405–409. https://doi.org/10.1177/026119290703500412
Oliveira Pinho F, Pinto Joazeiro P, Santos AR Jr (2021) Evaluation of the growth and differentiation of human fetal osteoblasts (hFOB) cells on demineralized bone matrix (DBM). Organogenesis 17:136–149. https://doi.org/10.1080/15476278.2021.2003134
Pacaccio DJ, Stern SF (2005) Demineralized bone matrix: basic science and clinical applications. Clin Podiatr Med Surg 22:599–606. https://doi.org/10.1016/j.cpm.2005.07.001
Pietrzak WS, Ali SN (2015) The extraction and measurement of bone morphogenetic protein 7 from bovine cortical bone as a function of particle size. J Craniofac Surg 26:296–299. https://doi.org/10.1097/SCS.0000000000001301
Pietrzak WS, Ali SN (2017) The elution kinetics of BMP-2, BMP-4, and BMP-7 from a commercial human demineralized bone matrix putty. J Craniofac Surg 28:2183–2188. https://doi.org/10.1097/SCS.0000000000004016
Pietrzak WS, Woodell-May J, McDonald N (2006) Assay of bone morphogenetic protein-2, -4, and -7 in human demineralized bone matrix. J Craniofac Surg 17:84–90. https://doi.org/10.1097/01.scs.0000179745.91165.73
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 12:81–88. https://doi.org/10.1007/s10561-009-9168-6
Pietrzak WS, Dow M, Gomez J, Soulvie M, Tsiagalis G (2012) The in vitro elution of BMP-7 from demineralized bone matrix. Cell Tissue Bank 13:653–661. https://doi.org/10.1007/s10561-011-9286-9
Schwartz Z, Somers A, Mellonig JT, Carnes DL Jr, Dean DD, Cochran DL et al (1998) Ability of commercial demineralized freeze-dried bone allograft to induce new bone formation is dependent on donor age but not gender. J Periodontol 69:470–478. https://doi.org/10.1902/jop.1998.69.4.470
Siclari VA, Zhu J, Akiyama K, Liu F, Zhang X, Chandra A et al (2013) Mesenchymal progenitors residing close to the bone surface are functionally distinct from those in the central bone marrow. Bone 53:575–586. https://doi.org/10.1016/j.bone.2012.12.013
van Houdt CIA, Cardoso DA, van Oirschot B, Ulrich DJO, Jansen JA, Leeuwenburgh SCG et al (2017) Porous titanium scaffolds with injectable hyaluronic acid-DBM gel for bone substitution in a rat critical-sized calvarial defect model. J Tissue Eng Regen Med 11:2537–2548. https://doi.org/10.1002/term.2151
Zhang H, Yang L, Yang XG, Wang F, Feng JT, Hua KC et al (2019) Demineralized bone matrix carriers and their clinical applications: an overview. Orthop Surg 11:725–737. https://doi.org/10.1111/os.12509
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We thank all the participants in this study.
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This study was funded entirely by the Spine Research Foundation. No funding was received for conducting this study.
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Author Yong-jie Zhao has received research support from Beijing Wonderful Biomaterials Company. Other authors have no competing interests to declare that are relevant to the content of this article.
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Zhao, Yj., Yin, G., Liu, B. et al. Variability of BMP-2 content in DBM products derived from different long bone. Cell Tissue Bank (2024). https://doi.org/10.1007/s10561-024-10132-5
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DOI: https://doi.org/10.1007/s10561-024-10132-5