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단백질 전달 영역 융합-Bone Morphogenetic Protein-2가 백서 두개골 결손부에서 골 조직 재생에 미치는 효과

Effect of protein transduction domain fused-bone morphogenetic protein-2 on bone regeneration in rat calvarial defects

최성호|김창성|조규성|김종관|채중규|엄유정
대한치주과학회 (The Korean Academy Of Periodontology)|2008.08|Journal of Periodontal & Implant Science|제38권 제2호|pp. 153-162

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키워드

Bone Regeneration; bone morphogenetic protein.
주제분류치의학

초록

Purpose: Recombining bone morphogenetic protein (BMP) is usually acquiredfrom high level animals. Though this method is effective, its high cost limits its use. The purpose of this study was to evaluate the effect of bone morphogenetic protein-2 with protein transduction domain (BMP-2/PTD;TATBMP-2) on bone regeneration. Rat calvarial defect model and osteoblastic differentiation model using MC3T3 cell were used for the purpose of the study.Materials and Methods: MC3T3 cells were cultured until they reached a confluence stage. The cells were treated with 0, 0.1, 1, 10, 100, 500ng/ml of BMP-2/PTD for 21 days and at the end of the treatment, osteoblastic differentiation was evaluated usingvon Kossa staining. An 8mm, calvarial, critical-size osteotomy defect was created in each of 48 male Spraque-Dawley rats (weight 250~300g). Three groups of 16 animals each received either BMP-2/PTD (0.05mg/ml) in a collagen carrier, collagen only, or negative surgical control. And each group was divided into 2 and 8 weeks healing intervals. The groups were evaluated by histologic analysis(8 animals/group/healing intervals)Result: In osteoblastic differentiation evaluation test, a stimulatory effect of BMP-2/PTD was observed in 10ng/ml of BMP-2/PTD with no observation of dose-dependent manner. The BMP-2/PTD group showed enhanced local bone formation in the rat calvarial defect at 2 weeks. New bone was observed at the defect margin and central area ofthe defect. However, new bone formation was observed only in 50% of animals used for 2weeks. In addition, there was no new bone formation observed at 8 weeks.Conclusion: The results of the present study indicated that BMP-2/PTD(TATBMP-2) have an positive effect on the bone formation in vitro and in vivo. However, further study should be conducted for the reproducibility of the outcomes.(J Korean Acad Periodontol 2008;38:153-162)

Abstract

Purpose: Recombining bone morphogenetic protein (BMP) is usually acquiredfrom high level animals. Though this method is effective, its high cost limits its use. The purpose of this study was to evaluate the effect of bone morphogenetic protein-2 with protein transduction domain (BMP-2/PTD;TATBMP-2) on bone regeneration. Rat calvarial defect model and osteoblastic differentiation model using MC3T3 cell were used for the purpose of the study.Materials and Methods: MC3T3 cells were cultured until they reached a confluence stage. The cells were treated with 0, 0.1, 1, 10, 100, 500ng/ml of BMP-2/PTD for 21 days and at the end of the treatment, osteoblastic differentiation was evaluated usingvon Kossa staining. An 8mm, calvarial, critical-size osteotomy defect was created in each of 48 male Spraque-Dawley rats (weight 250~300g). Three groups of 16 animals each received either BMP-2/PTD (0.05mg/ml) in a collagen carrier, collagen only, or negative surgical control. And each group was divided into 2 and 8 weeks healing intervals. The groups were evaluated by histologic analysis(8 animals/group/healing intervals)Result: In osteoblastic differentiation evaluation test, a stimulatory effect of BMP-2/PTD was observed in 10ng/ml of BMP-2/PTD with no observation of dose-dependent manner. The BMP-2/PTD group showed enhanced local bone formation in the rat calvarial defect at 2 weeks. New bone was observed at the defect margin and central area ofthe defect. However, new bone formation was observed only in 50% of animals used for 2weeks. In addition, there was no new bone formation observed at 8 weeks.Conclusion: The results of the present study indicated that BMP-2/PTD(TATBMP-2) have an positive effect on the bone formation in vitro and in vivo. However, further study should be conducted for the reproducibility of the outcomes.(J Korean Acad Periodontol 2008;38:153-162)

참고문헌 (43)

  1. Urist MR, "Bone: formation by autoinduction", Science, 150, pp.893~899 (1965)
  2. Wozney JM, "Novel regulators of bone formation: Molecular clones and activities", Science, 242, pp.1528~1534 (1988)
  3. Celeste AJ, "Identification of transforming growth factor-family members present in bone-inductive protein purified from bovine bone", Proc Natl Acad Sci U S A, 87, pp.9843~9847 (1990)
  4. Kim CS, "The effect of recombinant human bone morphogenetic protein-4 on the osteoblastic differentiation of mouse calvarial cells affected by Porphyromonas gingivalis", J Periodontol, 73, pp.1126~1132 (2002)
  5. Wikesjo UM, "Periodontal repair in dogs: effect of rhBMP-2 concentration on regeneration of alveolar bone and periodontal attachment", J Clin Periodontol, 26, pp.392~400 (1999)
  6. Kingsley DM, "The mouse short ear skeletal morphogenesis locus is associated with defects in a bone morphogenetic member of the TGF superfamily", Cell, 71, pp.399~410 (1992)
  7. Solloway MJ, "Mice lacking BMP-6 function", Dev Genet, 22, pp.321~323 (1998)
  8. Jena N, "BMP7 null mutation in mice: Developmental defects in skeleton, kidney, and eye", Exp Cell Res, 230, pp.28~37 (1997)
  9. Sampath TK, "Recombinant human osteo-genic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro", J Biol Chem, 267, pp.20352~20362 (1992)
  10. Wang EA, "Recombinant human bone morphogenetic protein induces bone formation", Proc Natl Acad Sci U S A, 87, pp.2220~2224 (1990)
  11. Cook SD, "Bone defect healing with an osteogenic protein-1 device combined with carboxymethylcellulose", J Biomed Mater Res Appl Biomater, 75, pp.137~145 (2005)
  12. Cook SD, "The effect of recombinant human osteogenic protein-1 on healing of large segmental bone defects", J Bone Joint Surg Am, 176, pp.827~838 (1994)
  13. Cook SD, "Effect of recombinant human osteogenic protein-1 on healing of segmental defects in non-human primates", J Bone Joint Surg Am, 77, pp.734~750 (1995)
  14. Reddi AH, "Bone Morphogenetic Proteins: From Basic Science to Clinical Applications", J Bone Joint Surg Am, 83, pp.S1~S6 (2001)
  15. Griffith DL, "Three- dimensional structure of recombinant human osteogenic protein 1 : Structural paradigm for the transforming growth factor superfamily", Proc Natl Acad Sci USA, 93, pp.878~883 (1996)
  16. Hong SJ, "Fibrin-Fibronectin Sealing System in Combination with -Tricalcium Phosphate as a Carrier for Recombinant Human Bone Morphogenetic Protein-2: Effects on Bone Formation in Rat Calvarial Defects", Biomaterials, 27, pp.3810~3816 (2005)
  17. Hyun SJ, "The effect of recombinant human bone morphogenetic protein-2, 4 and 7 on bone formation in rat calvarial defects", J Periodontol, 76, pp.1~8 (2005)
  18. Choi SH, "Effect of recombinant human bone morphogenetic protein-2", absorbable collagen sponge (rhBMP-2, J Periodontol, pp.2002, 73[DOI]
  19. Pang EK, "Effect of recombinant human bone morphogenetic protein-4 dose on bone formation in rat calvarial defects", J Periodontol, 75, pp.1364~1370 (2004)
  20. Ahn SH, "Effect of Recombinant Human Bone Morphogenetic Protein- 4 with Carriers in Rat Calvarial Defects", J Periodontol, 74, pp.787~797 (2003)
  21. Frankel AD, "Cellular uptake of the TAT protein from human immunodeficiency virus", Cell, 55, pp.1189~1193 (1988)
  22. Watson K, "HIV-1-trans-activating(TAT) protein both a target and a tool in therapeutic approaches", Biochem Pharmacol, 58, pp.1521~1528 (1999)
  23. Lindgren M, "Langel U. Cell penetrating peptides", Trends Pharmacol Sci, 21, pp.99~103 (2000)
  24. Schwarze SR, "Protein transduction : unrestricted delivery into all cell?", Trends cell Biol, 10, pp.290~295 (2000)
  25. Schwarze SR, "In vivo protein transduction : intercellular delivery of biological active proteins. componds and DNA", Trends Pharmacol Sci, 21, pp.45~48 (2000)
  26. Fawell S, "TAT- mediated delivery of heterologous proteins into cells", Proc Natl Acad Sci USA, 91, pp.664~668 (1994)
  27. Nagahara H, "Transduction of full- length TAT union proteins into mammalian cells: TAT -p27Kip1 induces cell migration", Nat Med, 4, pp.1449~1452 (1998)
  28. 정성원, "Protein transduction domain을 이용한 recombinant human bone morphogenetic protein-2의 골재생효과", Journal of Periodontal & Implant Science, 37(3), pp.497~509 (2007), 대한치주과학회
  29. Han DK, "Effect of a Fibrin-fibronectin sealing system as a carrier for recombinant human bone morphogenetic protein-4 on bone formation in rat calvarial defects", J Periodontol, 76, pp.2216~2222 (2005)
  30. Kim CS, "Ectopic bone formation associated with recombinanthuman bone morphogenetic protein-2 using absorbable collagen sponge and beta tricalcium phosphate as carriers", Biomaterials, 19, pp.1~7 (2004)
  31. Urist MR, "Beta-tricalcium phosphate delivery system for bone morphogenetic protein", Clin Orthop, 187, pp.277~280 (1984)
  32. Schmitz JP, "The critical size defect as an experimental model for craniomandibulofacial nonunion", Clin Orthop, 205, pp.299~308 (1986)
  33. King GN, "Factors that modulate the effect of bone morphogenetic protein-induced periodontal regeneration; a critical review", J Periodontol, 73, pp.925~936 (2002)
  34. Yasco AW, "The healing of segmental bone defects, induced by recombinant human bone morphogenetic protein (rhBMP-2)", J Bone Joint Surg Am, 74-A, pp.659~671 (1992)
  35. Barboza EP, "Ridge augmentation following implantation of recombinant human bone morphogenetic protein-2 in the dog", J Periodontol, 71, pp.488~496 (2000)
  36. Frankel AD, "Cellular uptake of the TAT protein from human immunodeficiency virus", Cell, 55, pp.1189~1193 (1988)
  37. Maurice Green, "Loewenstein. Autonomous Functional Domains of Chemically Synthesized Human Immunodeficiency Virus Tat Trans-Activator Protein", Cell, 55, pp.1179~1188 (1988)
  38. Ford KG, "Farzaneh F. Protein transduction: an alternative to genetic intervention?", Gene therapy, 8, pp.1~4 (2001)
  39. Leifert LA, "Translocatory proteins” and “protein transduction domains”: a critical analysis of their biological effects and the underlying mechanism", Mol Ther, 8, pp.13~20 (2003)
  40. Ozkaynak E, "Op-1 cDNA encodes an osteogenic protein in the TGF-beta family", EMBO J, 9, pp.2085~2093 (1990)
  41. Takakis DN, "Peri-implant bone regenerationusing recombinant human bone morphogenetic protein-2 in a canine model: A dose-response study", J Periodontal Res, 37, pp.93~100 (2002)
  42. Sandhu HS, "Effective doses of recombinant human bone morphogenetic protein-2 in experimental spinal fusion", Spine, 21, pp.2115~2122 (1996)
  43. Jehangir SW, "Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis", Nature medicine, 10, pp.311~315 (2004)

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