Abdolhamid MEIMANDI PARIZI, Ahmad ORYAN, Shahram HADDADI, Amin BIGHAM SADEGH

Histopathological and biomechanical evaluation of bone healing properties of DBM and DBM-G90 in a rabbit model

Objective: The present study was designed to investigate the effects of DBM and DBM-G90 on bone healing in a rabbit model.Methods: Thirty male white albino rabbits were used in this study. An incision was made in all rabbits under general anesthesia directly over the radius in order to expose it. A 10-mm segmental defect was created on the middle portion of each radius. The defects of 10 rabbits (Group I) were filled with DBM Block and Strip (Zimmer, Inc., Warsaw, IN, USA), the defects of 10 rabbits (Group II) were filled with DBM soaked in G90, and the defects of 10 rabbits (Group III/control) were left empty. The rabbits were euthanized at 60 days postoperatively for histopathological and biomechanical evaluation.Results: At the histopathologic level, the defects of the animals in the DBM and DBM-G90 groups showed more advanced healing criteria than those of the control group. In biomechanical findings, there was a statistically significant difference between the injured bones and contralateral normal bones of the control group in terms of measured strength. There was not a statistically significant difference between the treated bones of the DBM and DBM-G90 groups with contralateral normal bones, nor was there a statistically significant difference between the treated bones of the DBM and DBM-G90 groups with the treated bones of the control group, in terms of other biomechanical tests.Conclusion: Based on the histopathological and biomechanical findings, the DBM and DBM-G90 groups demonstrated superior osteogenic potential; however, G90 shows no superiority over DBM on bone healing.

___

Shafiei Z, Bigham AS, Dehghani SN, Nezhad ST. Fresh cortical autograft versus fresh cortical allograft effects on experimental bone healing in rabbits: radiological, histo- pathological and biomechanical evaluation. Cell Tissue Bank 2009;10:19-26.

Bigham-Sadegh A, Oryan A. Selection of animal models for pre-clinical strategies in evaluating the fracture heal- ing, bone graft substitutes and bone tissue regeneration and engineering. Connect Tissue Res 2015;56:175-94.

Bigham-Sadegh A, Karimi I, Alebouye M, Shafie-Sarvest- ani Z, Oryan A. Evaluation of bone healing in canine tibial defects filled with cortical autograft, commercial-DBM, calf fetal DBM, omentum and omentum-calf fetal DBM. J Vet Sci 2013;14:337-43.

Stevenson S. Enhancement of fracture healing with autog- enous and allogeneic bone grafts. Clin Orthop Relat Res 1998;(355 Suppl):239-46.

Banwart JC, Asher MA, Hassanein RS. Iliac crest bone graft harvest donor site morbidity. A statistical evaluation. Spine (Phila Pa 1976) 1995;20:1055-60.

Bigham A, Shadkhast M, Dehghani S. Autogenous bone marrow concurrent with static magnetic field effects on bone-defect healing: radiological and histological study. Comparative Clinical Pathology 2009;18:163-8.

Parizi AM, Oryan A, Shafiei-Sarvestani Z, Bigham AS. Human platelet rich plasma plus Persian Gulf coral effects on experimental bone healing in rabbit model: radiologi- cal, histological, macroscopical and biomechanical evalua- tion. J Mater Sci Mater Med 2012;23:473-83.

Oryan A, Meimandi Parizi A, Shafiei-Sarvestani Z, Big- ham AS. Effects of combined hydroxyapatite and human platelet rich plasma on bone healing in rabbit model: ra- diological, macroscopical, hidtopathological and biome- chanical evaluation. Cell Tissue Bank 2012;13:639-51.

Bigham-Sadegh A, Mirshokraei P, Karimi I, Oryan A, Aparviz A, Shafiei-Sarvestani Z. Effects of adipose tis- sue stem cell concurrent with greater omentum on ex- perimental long-bone healing in dog. Connect Tissue Res 2012;53:334-42.

Bigham AS, Dehghani SN, Shafiei Z, Torabi Nezhad S. Xenogenic demineralized bone matrix and fresh autog- enous cortical bone effects on experimental bone healing: radiological, histopathological and biomechanical evalua- tion. J Orthop Traumatol 2008;9:73-80.

Du C, Cui FZ, Feng QL, Zhu XD, de Groot K. Tis- sue response to nano-hydroxyapatite/collagen com- posite implants in marrow cavity. J Biomed Mater Res 1998;42:540-8.

El-Ghannam A. Bone reconstruction: from bioceramics to tissue engineering. Expert Rev Med Devices 2005;2:87- 101.

Li J, Lin Z, Zheng Q, Guo X, Lan S, Liu S, et al. Repair of rabbit radial bone defects using true bone ceramics com- bined with BMP-2-related peptide and type I collagen. Materials science and engineering: C 2010;30:1272-9.

McAuliffe JA. Bone graft substitutes. Journal of Hand Therapy 2003;16:180-187.

Wozney JM, Seeherman HJ. Protein-based tissue engi- neering in bone and cartilage repair. Curr Opin Biotechnol 2004;15:392-8.

Jin DD. Bone matrix gelatin. Clinical application in 38 cases. [Article in Chinese] Zhonghua Wai Ke Za Zhi 1991;29:312-4, 335. [Abstract]

Riley EH, Lane JM, Urist MR, Lyons KM, Lieberman JR. Bone morphogenetic protein-2: biology and applications. Clin Orthop Relat Res 1996;324:39-46.

Bostrom MP, Lane JM, Berberian WS, Missri AA, Tomin E, Weiland A, et al. Immunolocalization and expression of bone morphogenetic proteins 2 and 4 in fracture healing. J Orthop Res 1995;13:357-67.

Cook SD, Baffes GC, Wolfe MW, Sampath TK, Rueger DC, Whitecloud TS 3rd. The effect of recombinant human osteogenic protein-1 on healing of large segmental bone defects. J Bone Joint Surg Am 1994;76:827-38.

Kirker-Head CA. Recombinant bone morphogenetic pro- teins: novel substances for enhancing bone healing. Vet Surg 1995;24:408-19.

Reddi AH. Bone morphogenetic proteins, bone mar- row stromal cells, and mesenchymal stem cells. Maureen Owen revisited. Clin Orthop Relat Res 1995;313:115-9.

Loredo GA, MacDonald MH, Benton HP. Regulation of glycosaminoglycan metabolism by bone morphogenetic protein-2 in equine cartilage explant cultures. Am J Vet Res 1996;57:554-9.

Tanaka T, Fujii K, Ohta M, Soshi S, Kitamura A, Murota K. Use of a guanidine extract of demineralized bone in the treatment of osteochondral defects of articular cartilage. J Orthop Res 1995;13:464-9.

El-Kamali HH. Folk medicinal use of some animal prod- ucts in Central Sudan. J Ethnopharmacol 2000;72:279-82.

Hrzenjak T, Hrzenjak M, Kasuba V, Efenberger- Marinculi? P, Levanat S. A new source of biologically ac- tive compounds--earthworm tissue (Eisenia foetida, Lum- bricus rubelus). Comp Biochem Physiol Comp Physiol 1992;102:441-7.

Popovi? M, Hrcenjak TM, Babi? T, Kos J, Grdisa M. Ef- fect of earthworm (G-90) extract on formation and lysis of clots originated from venous blood of dogs with car- diopathies and with malignant tumors. Pathol Oncol Res 2001;7:197-202.

Popovi? M, Hrzenjak T, Grdisa M, Vukovi? S. Adhesins of immunoglobulin-like superfamily from earthworm Eisenia foetida. Gen Pharmacol 1998;30:795-800.

Hrzenjak M, Kobrehel D, Levanat S, Jurin M, Hrzenjak T. Mitogenicity of the earthworm's (Eisenia foetida) insulin- like proteins. Comp Biochem Physiol B 1993;104:723-9.

Hrzenjak T, Popovi? M, Bozi? T, Grdisa M, Kobrehel D, Tiska-Rudman L. Fibrinolytic and anticoagulative activi- ties from the earthworm Eisenia foetida. Comp Biochem Physiol B Biochem Mol Biol 1998;119:825-32.

Grdisa M, Popovic M, Hrzenjak T. Glycolipoprotein ex- tract (G-90) from earthworm Eisenia foetida exerts some antioxidative activity. Comp Biochem Physiol A Mol In- tegr Physiol 2001;128:821-5.

Vail TB, Trotter GW, Powers BE. Equine demineralized bone matrix: relationship between particle size and osteo- induction. Vet Surg 1994;23:386-95.

Forell EB, Straw RC, Powers BE, Johnson J, Cooper MF, St JW. Evaluation of the osteoinductive capacity of canine demineralized bone matrix in heterotopic muscle sites of athymic rats. Vet com Orthop traumatol 1993;6:25-32.

Reddi AH, Huggins C. Biochemical sequences in the transformation of normal fibroblasts in adolescent rats. Proc Natl Acad Sci U S A 1972;69:1601-5.

Urist MR, Strates BS. Bone formation in implants of partially and wholly demineralized bone matrix. Includ- ing observations on acetone-fixed intra and extracellular proteins. Clin Orthop Relat Res 1970;71:271-8.

Bolander ME, Balian G. The use of demineralized bone matrix in the repair of segmental defects. Augmentation with extracted matrix proteins and a comparison with au- tologous grafts. J Bone Joint Surg Am 1986;68:1264-74.

Matos MA, Araşjo FP, Paixão FB. Histomorphometric evaluation of bone healing in rabbit fibular osteotomy model without fixation. J Orthop Surg Res 2008;3:4.

Shafiei-Sarvestani Z, Oryan A, Bigham AS, Meimandi- Parizi A. The effect of hydroxyapatite-hPRP, and coral- hPRP on bone healing in rabbits: radiological, biome- chanical, macroscopic and histopathologic evaluation. Int J Surg 2012;10:96-101.

Bigham AS, Dehghani SN, Shafiei Z, Nezhad ST. Experi- mental bone defect healing with xenogenic demineralized bone matrix and bovine fetal growth plate as a new xe- nograft: radiological, histopathological and biomechanical evaluation. Cell Tissue Bank 2009;10:33-41.

Landoni MF. Management of pain in laboratory animals In: Rigalli A,Elina Di Loreto V, eds. Experimental surgical models in the laboratory rat. 1 ed. Broken Sound Parkway NW: CRC Press, 2009. p. 17-20.

Emery SE, Brazinski MS, Koka A, Bensusan JS, Steven- son S. The biological and biomechanical effects of irra- diation on anterior spinal bone grafts in a canine model. J Bone Joint Surg Am 1994;76:540-8.

Oryan A, Goodship AE, Silver IA. Response of a colla- genase-induced tendon injury to treatment with a poly- sulphated glycosaminoglycan (Adequan). Connect Tissue Res 2008;49:351-60.

Oryan A, Moshiri A, Meimandiparizi AH. Effects of sodium-hyaluronate and glucosamine-chondroitin sul- fate on remodeling stage of tenotomized superficial digital flexor tendon in rabbits: a clinical, histopathological, ultra- structural, and biomechanical study. Connect Tissue Res 2011;52:329-39.

Oryan A, Shoushtari AH. Biomechanical properties and dry weight content of the developing superficial digital flexor tendon. Comp Clin Pathol 2009;18:131-7.

Wang KH, Brose K, Arnott D, Kidd T, Goodman CS, Henzel W, et al. Biochemical purification of a mammalian slit protein as a positive regulator of sensory axon elonga- tion and branching. Cell 1999;96:771-84.

Oryan A, Alidadi S, Moshiri A, Maffulli N. Bone regener- ative medicine: classic options, novel strategies, and future directions. J Orthop Surg Res 2014;9:18.

Urist MR, Mikulski A, Lietze A. Solubilized and insolubi- lized bone morphogenetic protein. Proc Natl Acad Sci U S A 1979;76:1828-32.

Urist MF, Sato K, Brownell AG. Human bone morphoge- netic protein. Proceedings of the Society for Experimental Biology and Medicine Society for Experimental Biology and Medicine (New York, NY) 1983;194-9.

Martin GJ Jr, Boden SD, Titus L, Scarborough NL. New formulations of demineralized bone matrix as a more effec- tive graft alternative in experimental posterolateral lumbar spine arthrodesis. Spine (Phila Pa 1976) 1999;24:637-45.

Guizzardi S, Di Silvestre M, Scandroglio R, Ruggeri A, Savini R. Implants of heterologous demineralized bone matrix for induction of posterior spinal fusion in rats. Spine (Phila Pa 1976) 1992;17:701,7.

Omar HE-DM, Ibraheim ZZ, El-Shimy NA, Ali RS. Anti-inflammatory, antipyretic and antioxidant activities of the earthworms extract. Journal of Biology and Earth Sciences 2012;2:10-7.

Balamurugan M, Parthasarathi K, Cooper EL, Rangana- than LS. Anti-inflammatory and anti-pyretic activities of earthworm extract-Lampito mauritii (Kinberg). J Ethno- pharmacol 2009;121:330-2.

Kini U, Nandeesh BN. Physiology of bone formation, re- modeling and metabolism In: Fogelman I, Gnanasegaran G, Van der Wall H, eds. Radionuclide and Hybrid Bone Imaging: Springer 2012. p. 29-57.

Grdisa M, Popovi? M, Hrzenjak T. Stimulation of growth factor synthesis in skin wounds using tissue extract (G- 90) from the earthworm Eissenia foetida. Cell Biochem Funct 2004;22:373-8.

Popovic M, Grdisa M, Hrzenjak TM. Glycolipoprotein G-90 obtained from the earthworm Eisenia foetida exerts antibacterial activity. Vet Arhiv 2005;75:119-28.