Vol 79, No 3 (2020)
Original article
Published online: 2019-09-03
Effects of alloplastic graft material combined with a topical ozone application on calvarial bone defects in rats
Abstract
Background: This study presents the evaluation of the damage in the bone tissue resulting from a calvarial defect in rats and the efficiency of exposure to an ozone application with an alloplastic bone graft on the calvarial bone damage.
Materials and methods: Wistar male rats (n = 56) were divided into four groups: a control group (n = 14), defect and ozone group (n = 14), defect and graft group (n = 14), and defect, graft, and ozone group (n = 14). Under anaesthesia, a circular full-thickness bone defect was created in all groups, and the experimental groups were further divided into two sub-groups, with 7 rats in each group sacrificed at the end of the 4th and 8th weeks. Bone samples were dissected, fixed in 10% formalin solution, and decalcified with 5% ethylene-diamine-tetraacetic acid (EDTA). After the routine follow-up on tissues, immunostaining of osteopontin and osteonectin antibodies was applied to sections and observed under a light microscope.
Results: The control group exhibited osteopontin and osteonectin expression in fibroblasts and inflammatory cells at the end of the 4th week with an acceleration at the 8th week. Ozone administration elucidated new trabecular bone formation by increasing osteoblastic activity. Lastly, our observations underscore that a combination of allograft and ozone application increased the osteoblast, osteocyte, and bone matrix development at the 4th and 8th weeks.
Conclusions: Exposure to an ozone application with an alloplastic bone graft on calvarial bone damage may induce osteoblastic activity, matrix development, mature bone cell formation, and new bone formation in rats.
Keywords: calvarial defectsosteopontinosteonectinratimmunohistochemistry
References
- Alpan AL, Toker H, Ozer H. Ozone therapy enhances osseous healing in rats with diabetes with calvarial defects: a morphometric and immunohistochemical study. J Periodontol. 2016; 87(8): 982–989.
- An YZ, Heo YK, Lee JS, et al. Dehydrothermally cross-linked collagen membrane with a bone graft improves bone regeneration in a rat calvarial defect model. Materials (Basel). 2017; 10(8).
- Ardeshirylajimi A, Farhadian S, Adegani FJ, et al. Enhanced osteoconductivity of polyethersulphone nanofibres loaded with bioactive glass nanoparticles in in vitro and in vivo models. Cell Prolif. 2015; 48(4): 455–464.
- Artas G, Gul M, Acikan I, et al. A comparison of different bone graft materials in peri-implant guided bone regeneration. Braz Oral Res. 2018; 32: e59.
- Bayer S, Kazancioglu HO, Acar AH, et al. Comparison of laser and ozone treatments on oral mucositis in an experimental model. Lasers Med Sci. 2017; 32(3): 673–677.
- Beirne OR. Comparison of complications after bone removal from lateral and medial plates of the anterior ileum for mandibular augmentation. Int J Oral Maxillofac Surg. 1986; 15(3): 269–272.
- Bodde EWH, Spauwen PHM, Mikos AG, et al. Closing capacity of segmental radius defects in rabbits. J Biomed Mater Res A. 2008; 85(1): 206–217.
- Buffoli B, Favero G, Borsani E, et al. Sodium-DNA for bone tissue regeneration: an experimental study in rat calvaria. Biomed Res Int. 2017; 2017: 7320953.
- Buyuk SK, Ramoglu SI, Sonmez MF. The effect of different concentrations of topical ozone administration on bone formation in orthopedically expanded suture in rats. Eur J Orthod. 2016; 38(3): 281–285.
- Byrd HS, Hobar PC, Shewmake K. Augmentation of the craniofacial skeleton with porous hydroxyapatite granules. Plast Reconstr Surg. 1993; 91(1): 15–22; discussion 23.
- Caliaperoumal G, Souyet M, Bensidhoum M, et al. Type 2 diabetes impairs angiogenesis and osteogenesis in calvarial defects: MicroCT study in ZDF rats. Bone. 2018; 112: 161–172.
- Cancedda R, Giannoni P, Mastrogiacomo M. A tissue engineering approach to bone repair in large animal models and in clinical practice. Biomaterials. 2007; 28(29): 4240–4250.
- Cestari TM, Granjeiro JM, de Assis GF, et al. Bone repair and augmentation using block of sintered bovine-derived anorganic bone graft in cranial bone defect model. Clin Oral Implants Res. 2009; 20(4): 340–350.
- Choi ST, Kim JH, Kang EJ, et al. Osteopontin might be involved in bone remodelling rather than in inflammation in ankylosing spondylitis. Rheumatology (Oxford). 2008; 47(12): 1775–1779.
- Chou J, Komuro M, Hao J, et al. Bioresorbable zinc hydroxyapatite guided bone regeneration membrane for bone regeneration. Clin Oral Implants Res. 2016; 27(3): 354–360.
- Corsair A. A clinical evaluation of resorbable hydroxylapatite for the repair of human intra-osseous defects. J Oral Implantol. 1990; 16(2): 125–128.
- Deng Y, Guo T, Li J, et al. Repair of calvarial bone defect using jarid1a-knockdown bone mesenchymal stem cells in rats. Tissue Eng Part A. 2018; 24(9-10): 711–718.
- Diker N, Gulsever S, Koroglu T, et al. Effects of hyaluronic acid and hydroxyapatite/beta-tricalcium phosphate in combination on bone regeneration of a critical-size defect in an experimental model. J Craniofac Surg. 2018; 29(4): 1087–1093.
- Dova S, Karkos PD, Constantinidis J. Reconstruction of frontal defects with calvarial grafts. Rhinology. 2018; 56(3): 297–302.
- Duman I, Davul S, Gokce H, et al. Effects of gaseous ozone treatment on bone regeneration in femoral defect model in rats. J Hard Tissue Biol. 2017; 26(1): 7–12.
- Erdemci F, Gunaydin Y, Sencimen M, et al. Histomorphometric evaluation of the effect of systemic and topical ozone on alveolar bone healing following tooth extraction in rats. Int J Oral Maxillofac Surg. 2014; 43(6): 777–783.
- Erdmann N, Bondarenko A, Hewicker-Trautwein M, et al. Evaluation of the soft tissue biocompatibility of MgCa0.8 and surgical steel 316L in vivo: a comparative study in rabbits. Biomed Eng Online. 2010; 9: 63.
- Ferland CE, Laverty S, Beaudry F, et al. Gait analysis and pain response of two rodent models of osteoarthritis. Pharmacol Biochem Behav. 2011; 97(3): 603–610.
- Ferrari JD, Bach BR. Bone graft procurement for patellar defect grafting in anterior cruciate ligament reconstruction. Arthroscopy. 1998; 14(5): 543–545.
- Fujishiro T, Bauer TW, Kobayashi N, et al. Histological evaluation of an impacted bone graft substitute composed of a combination of mineralized and demineralized allograft in a sheep vertebral bone defect. J Biomed Mater Res A. 2007; 82(3): 538–544.
- Garg AK. Grafting Materials in Repair and Restoration. In: Lynch SE, Genco RJ, Marx RE (ed. Tissue Engineering: Applications in Maxillofacial Surgery and Periodontics, Illinois: Quintessence Publishing. 1999: 83–102.
- Hou R, Chen F, Yang Y, et al. Comparative study between coral-mesenchymal stem cells-rhBMP-2 composite and auto-bone-graft in rabbit critical-sized cranial defect model. J Biomed Mater Res A. 2007; 80(1): 85–93.
- Huang B, Sun Y, Maciejewska I, et al. Distribution of SIBLING proteins in the organic and inorganic phases of rat dentin and bone. Eur J Oral Sci. 2008; 116(2): 104–112.
- Ihara H, Denhardt DT, Furuya K, et al. Parathyroid hormone-induced bone resorption does not occur in the absence of osteopontin. J Biol Chem. 2001; 276(16): 13065–13071.
- Isler SC, Unsal B, Soysal F, et al. The effects of ozone therapy as an adjunct to the surgical treatment of peri-implantitis. J Periodontal Implant Sci. 2018; 48(3): 136–151.
- Jakus AE, Rutz AL, Jordan SW, et al. Hyperelastic "bone": A highly versatile, growth factor-free, osteoregenerative, scalable, and surgically friendly biomaterial. Sci Transl Med. 2016; 8(358): 358ra127.
- Kan B, Sencimen M, Bayar GR, et al. Histomorphometric and microtomographic evaluation of the effects of hyperbaric oxygen and systemic ozone, used alone and in combination, on calvarial defect healing in rats. J Oral Maxillofac Surg. 2015; 73(6): 1231.e1–1231.10.
- Kazancioglu HO, Ezirganli S, Aydin MS. Effects of laser and ozone therapies on bone healing in the calvarial defects. J Craniofac Surg. 2013; 24(6): 2141–2146.
- Kelm RJ, et al. Jr, Swords NA, Orfeo T, Osteonectin in matrix remodeling. A plasminogen-osteonectin collagen complex. J Biol Chem. 1994; 269(48): 30147–30153.
- Khojasteh A, Eslaminejad MB, Nazarian H. Mesenchymal stem cells enhance bone regeneration in rat calvarial critical size defects more than platelete-rich plasma. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2008; 106(3): 356–62; discussion 363.
- Kim KS, Lee JY, Kang YM, et al. Small intestine submucosa sponge for in vivo support of tissue-engineered bone formation in the presence of rat bone marrow stem cells. Biomaterials. 2010; 31(6): 1104–1113.
- Klein-Nulend J, Roelofsen J, Semeins CM, et al. Mechanical stimulation of osteopontin mRNA expression and synthesis in bone cell cultures. J Cell Physiol. 1997; 170(2): 174–181, doi: 10.1002/(SICI)1097-4652(199702)170:2<174::AID-JCP9>3.0.CO;2-L.
- Koparal M, Irtegün S, Alan H, et al. Effects of Melatonin on Tibia Bone Defects in Rats. Int J Morphol. 2016; 34(2): 763–769.
- Laçin N, Kaya B, Deveci E, et al. Comparative evaluation of ozone treatment in critical size bone defects reconstructed with alloplastic bone grafts. Int J Clin Med. 2018; 09(07): 566–579.
- Lee MK, DeConde AS, Lee M, et al. Biomimetic scaffolds facilitate healing of critical-sized segmental mandibular defects. Am J Otolaryngol. 2015; 36(1): 1–6.
- Malard O, Guicheux J, Bouler JM, et al. Calcium phosphate scaffold and bone marrow for bone reconstruction in irradiated area: a dog study. Bone. 2005; 36(2): 323–330.
- Miron RJ, Zhang Q, Sculean A, et al. Osteoinductive potential of 4 commonly employed bone grafts. Clin Oral Investig. 2016; 20(8): 2259–2265.
- Ono N, Nakashima K, Rittling SR, et al. Osteopontin negatively regulates parathyroid hormone receptor signaling in osteoblasts. J Biol Chem. 2008; 283(28): 19400–19409.
- Ozdemir B, Kurtis B, Tuter G, et al. Osteocalcin and osteonectin expression after double application of platelet-rich plasma in rabbits. J Istanb Univ Fac Dent. 2016; 50(2): 1–9.
- Ozdemir H, Toker H, Balcı H, et al. Effect of ozone therapy on autogenous bone graft healing in calvarial defects: a histologic and histometric study in rats. J Periodontal Res. 2013; 48(6): 722–726.
- Ozeç I, Kiliç E, Gümüş C, et al. Effect of local simvastatin application on mandibular defects. J Craniofac Surg. 2007; 18(3): 546–550.
- Page D, Laskin D. Tissue response at the bone-implant interface in a hydroxylapatite augmented mandibular ridge. J Oral Maxillofac Surg. 1987; 45(4): 356–358.
- Pal U, Mishra N. Placement of implants in an ossifying fibroma defect obliterated with demineralized, freeze-dried bone allograft and Plasma-rich growth factor. Contemp Clin Dent. 2012; 3(4): 471–474.
- Pati F, Song TH, Rijal G, et al. Ornamenting 3D printed scaffolds with cell-laid extracellular matrix for bone tissue regeneration. Biomaterials. 2015; 37: 230–241.
- Pearce AI, Richards RG, Milz S, et al. Animal models for implant biomaterial research in bone: a review. Eur Cell Mater. 2007; 13: 1–10.
- Qin C, Baba O, Butler WT. Post-translational modifications of sibling proteins and their roles in osteogenesis and dentinogenesis. Crit Rev Oral Biol Med. 2004; 15(3): 126–136.
- Rasouli-Ghahroudi AA, Rokn A, Abdollahi M, et al. Effect of semelil, an herbal selenium-based medicine, on new bone formation in calvarium of rabbits. Biomed Res Int. 2018; 2018: 2860367.
- Reichert JC, Epari DR, Wullschleger ME, et al. Establishment of a preclinical ovine model for tibial segmental bone defect repair by applying bone tissue engineering strategies. Tissue Eng Part B Rev. 2010; 16(1): 93–104.
- Roach HI. Why does bone matrix contain non-collagenous proteins? The possible roles of osteocalcin, osteonectin, osteopontin and bone sialoprotein in bone mineralisation and resorption. Cell Biol Int. 1994; 18(6): 617–628.
- Rodriguez R, Kondo H, Nyan M, et al. Implantation of green tea catechin α-tricalcium phosphate combination enhances bone repair in rat skull defects. J Biomed Mater Res B Appl Biomater. 2011; 98(2): 263–271.
- Roux BM, Akar B, Zhou W, et al. Preformed vascular networks survive and enhance vascularization in critical sized cranial defects. Tissue Eng Part A. 2018; 24(21-22): 1603–1615.
- Sagai M, Bocci V. Mechanisms of action involved in ozone therapy: is healing induced via a mild oxidative stress? Med Gas Res. 2011; 1: 29.
- Saito A, Ooki A, Nakamura T, et al. Targeted reversion of induced pluripotent stem cells from patients with human cleidocranial dysplasia improves bone regeneration in a rat calvarial bone defect model. Stem Cell Res Ther. 2018; 9(1): 12.
- Sakata Y, Ueno T, Kagawa T, et al. Osteogenic potential of cultured human periosteum-derived cells - a pilot study of human cell transplantation into a rat calvarial defect model. J Craniomaxillofac Surg. 2006; 34(8): 461–465.
- Sodek J, Batista Da Silva AP, Zohar R. Osteopontin and mucosal protection. J Dent Res. 2006; 85(5): 404–415.
- Sodek J, Ganss B, McKee MD. Osteopontin. Crit Rev Oral Biol Med. 2000; 11(3): 279–303.
- Spicer PP, Kretlow JD, Young S, et al. Evaluation of bone regeneration using the rat critical size calvarial defect. Nat Protoc. 2012; 7(10): 1918–1929.
- Standal T, Borset M, Sundan A. Role of osteopontin in adhesion, migration, cell survival and bone remodeling. Exp Oncol. 2004; 26(3): 179–184.
- Toita R, Tsuru K, Ishikawa K. A superhydrophilic titanium implant functionalized by ozone gas modulates bone marrow cell and macrophage responses. J Mater Sci Mater Med. 2016; 27(8): 127.
- Thurner PJ, Chen CG, Ionova-Martin S, et al. Osteopontin deficiency increases bone fragility but preserves bone mass. Bone. 2010; 46(6): 1564–1573.
- Tricot M, Deleu PA, Detrembleur C, et al. Clinical assessment of 115 cases of hindfoot fusion with two different types of graft: allograft+dbm+bone marrow aspirate versus autograft+DBM. Orthop Traumatol Surg Res. 2017; 103(5): 697–702.
- Tshamala M, van Bree H. Osteoinductive properties of the bone marrow--myth or reality. Vet Comp Orthop Traumatol. 2006; 19(3): 133–141.
- Tye CE, Rattray KR, Warner KJ, et al. Delineation of the hydroxyapatite-nucleating domains of bone sialoprotein. J Biol Chem. 2003; 278(10): 7949–7955.
- Wang X, Zeng D, Weng W, et al. Alendronate delivery on amino modified mesoporous bioactive glass scaffolds to enhance bone regeneration in osteoporosis rats. Artif Cells Nanomed Biotechnol. 2018; 46(sup2): 171–181.
- Wang YH, Wu JY, Kong SC, et al. Low power laser irradiation and human adipose-derived stem cell treatments promote bone regeneration in critical-sized calvarial defects in rats. PLoS One. 2018; 13(4): e0195337.
- Yang Y, Cui Q, Sahai N. How does bone sialoprotein promote the nucleation of hydroxyapatite? A molecular dynamics study using model peptides of different conformations. Langmuir. 2010; 26(12): 9848–9859.
- Yang Yi, Hallgrimsson B, Putnins EE. Craniofacial defect regeneration using engineered bone marrow mesenchymal stromal cells. J Biomed Mater Res A. 2011; 99(1): 74–85.
