Bone Healing in Ovariectomized-rabbit Calvarial Defect with Tricalcium Phosphate Coated with Recombinant Human Bone Morphogenetic Protein-2 Genetically Engineered in Escherichia coli원문보기
Purpose: This study compares the bone formation ability of tricalcium phosphate (TCP) with and without recombinant human bone morphogenetic protein-2 (rhBMP-2) and assesses TCP as a carrier of rhBMP-2. Methods: Bilateral round defects (diameter: 8.0 mm) were formed in the cranium of eight New Zealan...
Purpose: This study compares the bone formation ability of tricalcium phosphate (TCP) with and without recombinant human bone morphogenetic protein-2 (rhBMP-2) and assesses TCP as a carrier of rhBMP-2. Methods: Bilateral round defects (diameter: 8.0 mm) were formed in the cranium of eight New Zealand white rabbits. The defects were grafted with TCP only (control group) or with rhBMP-2-coated TCP (experimental group). The animals were sacrificed at 1st week, 2nd week, 4th week, and 8th week postoperatively; two rabbits sacrificed each time. The skulls were harvested and subjected to radiographic and histological examination. Results: Radiologic evaluation showed faster bone remodeling in the experimental group than in the control group. Histologic evaluation (H&E, Masson's trichrome stain) showed rapid bone formation, remodeling and calcification in the 1st and 2nd week in the experimental group. Immunohistochemical evaluation showed higher expression rate of osteoprotegerin, receptor activator of nuclear factor ${\kappa}B$ ligand, and receptor activator of nuclear factor ${\kappa}B$ in the experimental group at the 1st and 2nd week than in the control group. Conclusion: rhBMP-2 coated TCP resulted in rapid bone formation, remodeling, and calcification due to rhBMP-2's osteogenic effect. TCP performed properly as a carrier for rhBMP-2. Thus, the use of an rhBMP-2 coating on TCP had a synergic effect on bone healing and, especially, bone remodeling and maturation.
Purpose: This study compares the bone formation ability of tricalcium phosphate (TCP) with and without recombinant human bone morphogenetic protein-2 (rhBMP-2) and assesses TCP as a carrier of rhBMP-2. Methods: Bilateral round defects (diameter: 8.0 mm) were formed in the cranium of eight New Zealand white rabbits. The defects were grafted with TCP only (control group) or with rhBMP-2-coated TCP (experimental group). The animals were sacrificed at 1st week, 2nd week, 4th week, and 8th week postoperatively; two rabbits sacrificed each time. The skulls were harvested and subjected to radiographic and histological examination. Results: Radiologic evaluation showed faster bone remodeling in the experimental group than in the control group. Histologic evaluation (H&E, Masson's trichrome stain) showed rapid bone formation, remodeling and calcification in the 1st and 2nd week in the experimental group. Immunohistochemical evaluation showed higher expression rate of osteoprotegerin, receptor activator of nuclear factor ${\kappa}B$ ligand, and receptor activator of nuclear factor ${\kappa}B$ in the experimental group at the 1st and 2nd week than in the control group. Conclusion: rhBMP-2 coated TCP resulted in rapid bone formation, remodeling, and calcification due to rhBMP-2's osteogenic effect. TCP performed properly as a carrier for rhBMP-2. Thus, the use of an rhBMP-2 coating on TCP had a synergic effect on bone healing and, especially, bone remodeling and maturation.
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가설 설정
(G) Microphotograph at 8 weeks after graft in control site. (H) Microphotograph at 8 weeks after graft in experimental site. GM, graft material; HB, host bone.
(G) Regenerating bone at 8 weeks after graft in control site. (H) Regenerating bone at 8 weeks after graft in experimental site. Black arrows indicate OPG antibody reaction.
1. In the radiological results, bone remodeled faster in the experimental group than the control group. The BMD estimation by μCT indicated especially high values for the experimental group at the 1st, 2nd, and 4th weeks.
제안 방법
Additional sections were mounted on glass slides for immunohistochemical analysis of the expressions of osteo-protegerin (OPG), the receptor activator of nuclear factor κB ligand (RANKL), and the receptor activator of nuclear factor κB (RANK) in the rabbit calvarial defects.
At each time point (1st week, 2nd week, 4th week, 8th week), the calvaria of two rabbits were scanned by micro-computed tomography (μCT; FLEXTM for flatform X-OTM; GMI, Northridge, CA, USA) horizontally, coronally and sagittally for measurement of the bone mineral density (BMD) at the 1st, 2nd, 4th, and 8th weeks (Fig. 2).
In this study, we grafted either rhBMP-2-coated TCP or TCP only onto calvaria defects in ovariectomized rabbits and evaluated the extents of bone formation and remodeling.
In this study, we grafted rhBMP-2-coated TCP and uncoated TCP onto left- and right-side defects to the skull of ovariectomized rabbits and evaluated the extents of bone formation and remodeling in radiological, μCT, histological and immunohistochemical analyses (OPG, RANKL, RANK).
Serial cross-sections (5 μm) were cut through the large diameter of the defect and stained with hematoxylin-eosin (H&E) and Masson’s trichrome, with which the cellular reaction and bone formation were evaluated.
The rabbits were sacrificed at the 1st, 2nd, 4th, and 8th weeks postoperatively, two each time. The skulls were harvested and subjected to radiographic and histological examination.
The rabbits were sacrificed at the 1st, 2nd, 4th, and 8th weeks postoperatively, two each time. The skulls were harvested and subjected to radiographic and histological examination. All of the experiments were conducted in accordance with the ethical committee of the Medical Research Institute of Dong-A University.
대상 데이터
Eight New Zealand adult female rabbits older than 6 months, weighing between 2.5 and 3.5 kg, were used in the study. The animals were housed individually in standard rabbit cages at an ambient temperature of 20oC under a 12/12 hour light/dark cycle.
성능/효과
2. In the histological results (H&E, Masson’s Trichrome stain), bone formed rapidly in the experimental group, with remodeling and calcification in the 1st and 2nd weeks.
3. In the immunohistochemical results, the OPG, RANKL, and RANK all expressed at higher rates in the experimental group at the 1st and 2nd weeks than the control group. This implies that osteoblasts and osteoclasts are more active at the early stages.
The experimental group’s expression of OPG, RANKL, and RANK were higher in the 1st and 2nd weeks but decreased to the level of the control group in the 4th and 8th weeks.
참고문헌 (37)
1. Bassett CA Herrmann I Influence of oxygen concentration and mechanical factors on differentiation of connective tissues in vitro Nature 1961 190 460 1 13687643
2. Goldberg VM Stevenson S Natural history of autografts and allografts Clin Orthop Relat Res 1987 225 7 16 3315383
3. Okubo Y Bessho K Fujimura K Osteoinduction by recombinant human bone morphogenetic protein-2 at intramuscular, intermuscular, subcutaneous and intrafatty sites Int J Oral Maxillofac Surg 2000 29 62 6 10691147
9. Murata M Inoue M Arisue M Kuboki Y Nagai N Carrier-dependency of cellular differentiation induced by bone morphogenetic protein in ectopic sites Int J Oral Maxillofac Surg 1998 27 391 6 9804206
10. Nagai N Qin CL Nagatsuka H Inoue M Ishiwari Y Age effects on ectopic bone formation induced by purified bone morphogenetic protein Int J Oral Maxillofac Surg 1999 28 143 50 10102399
11. Kamakura S Sasano Y Homma H Suzuki O Kagayama M Motegi K Implantation of octacalcium phosphate nucleates isolated bone formation in rat skull defects Oral Dis 2001 7 259 65 11575878
12. Boyne PJ Marx RE Nevins M A feasibility study evaluating rhBMP-2/absorbable collagen sponge for maxillary sinus floor augmentation Int J Periodontic Restor Dent 1997 17 11 25
13. Kenley R Marden L Turek T Jin L Ron E Hollinger JO Osseous regeneration in the rat calvarium using novel delivery systems for recombinant human bone morphogenetic protein-2 (rhBMP-2) J Biomed Mater Res 1994 28 1139 47 7829544
14. Yasko AW Lane JM Fellinger EJ Rosen V Wozney JM Wang EA The healing of segmental bone defects, induced by recombinant human bone morphogenetic protein (rhBMP-2). A radiographic, histological, and biomechanical study in rats J Bone Joint Surg Am 1992 74 659 70 1378056
15. Ogose A Kondo N Umezu H Histological assessment in grafts of highly purified beta-tricalcium phosphate (OSferion) in human bones Biomaterials 2006 27 1542 9 16165205
16. Damien CJ Parsons JR Bone graft and bone graft substitutes: a review of current technology and applications J Appl Biomater 1991 2 187 208 10149083
19. Ohgushi H Okumura M Tamai S Shors EC Caplan AI Marrow cell induced osteogenesis in porous hydroxyapatite and tricalcium phosphate: a comparative histomorphometric study of ectopic bone formation J Biomed Mater Res 1990 24 1563 70 2277053
20. St John KR Zardiackas LD Terry RC Teasdall RD Cooke SE Mitias HM Histological and electron microscopic analysis of tissue response to synthetic composite bone graft in the canine J Appl Biomater 1995 6 89 97 7640444
22. Hughes FJ Collyer J Stanfield M Goodman SA The effects of bone morphogenetic protein-2, -4, and -6 on differentiation of rat osteoblast cells in vitro Endocrinology 1995 136 2671 7 7750491
23. Higuchi T Kinoshita A Takahashi K Oda S Ishikawa I Bone regeneration by recombinant human bone morphogenetic protein-2 in rat mandibular defects. An experimental model of defect filling J Periodontol 1999 70 1026 31 10505805
24. Matin K Senpuku H Hanada N Ozawa H Ejiri S Bone regeneration by recombinant human bone morphogenetic protein-2 around immediate implants: a pilot study in rats Int J Oral Maxillofac Implants 2003 18 211 7 12705298
25. Kuboki Y Saito T Murata M Two distinctive BMP-carriers induce zonal chondrogenesis and membranous ossification, respectively; geometrical factors of matrices for cell-differentiation Connect Tissue Res 1995 32 219 26 7554920
26. Ripamonti U Ma S Reddi AH The critical role of geometry of porous hydroxyapatite delivery system in induction of bone by osteogenin, a bone morphogenetic protein Matrix 1992 12 202 12 1328828
30. Pang EK Im SU Kim CS Effect of recombinant human bone morphogenetic protein-4 dose on bone formation in a rat calvarial defect model J Periodontol 2004 75 1364 70 15562914
31. Yang X Fang J Luo Y Effects of recombinant human bone morphogenetic protein 2 and osteogenic agents on proliferation and differentiation of rat mesenchymal stem cells Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2007 21 140 4 17357460
32. Lundgren S Rosenquist JB Short term bone healing in calcium deficiency osteopenia and disuse osteopenia: experimental studies in adult rats Scand J Dent Res 1992 100 337 9 1465568
33. Cao T Shirota T Yamazaki M Ohno K Michi KI Bone mineral density in mandibles of ovariectomized rabbits Clin Oral Implants Res 2001 12 604 8 11737104
34. Kwan Tat S Padrines M Théoleyre S Heymann D Fortun Y IL-6, RANKL, TNF-alpha/IL-1: interrelations in bone resorption pathophysiology Cytokine Growth Factor Rev 2004 15 49 60 14746813
35. Theoleyre S Wittrant Y Tat SK Fortun Y Redini F Heymann D The molecular triad OPG/RANK/RANKL: involvement in the orchestration of pathophysiological bone remodeling Cytokine Growth Factor Rev 2004 15 457 75 15561602
37. Koide M Murase Y Yamato K Noguchi T Okahashi N Nishihara T Bone morphogenetic protein-2 enhances osteoclast formation mediated by interleukin-1alpha through upregulation of osteoclast differentiation factor and cyclooxygenase-2 Biochem Biophys Res Commun 1999 259 97 102 10334922
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