골 대체물의 종류가 BMP-7 유전자의 형질 전환된 자가 골수 간세포를 이용한 토끼 두개골 결손 복구에 미치는 영향 (BMP-7 유전자의 형질 전환된 자가 골수 간세포로 골 결손 복구 시 골 대체물의 영향)

Effect of Bone Substitutes on Repair of the Large Sized Defect on the Rabbit Calvarium Using BMP-7 Gene Engineered Bone Marrow Stromal Cells

  • 조재호 (인제대학교 의과대학 서울백병원 정형외과) ;
  • 하철웅 (영남대학교 의과대학 정형외과) ;
  • 송인환 (영남대학교 의과대학 해부학교실) ;
  • 김석영 (영남대학교 의과대학 해부학교실) ;
  • 안면환 (영남대학교 의과대학 정형외과)
  • Cho, Jae Ho (Department of Orthopaedic Surgery, Seoul Paik Hospital, Inje University) ;
  • Ha, Chul Wung (Department of Orthopaedic Surgery, College of Medicine, Yeungnam University) ;
  • Song, In-Whan (Deparment of Anatomy, College of Medicine, Yeungnam University) ;
  • Kim, Sukyoung (School of Materials Science and Engineering, Yeungnam University) ;
  • Ahn, Myun Whan (Department of Orthopaedic Surgery, College of Medicine, Yeungnam University)
  • 발행 : 2012.06.01

초록

The aim of this study is to examine the efficacy of bone substitutes on repair of the large sized defect on the rabbit calvarium using BMP-7 gene engineered bone marrow stromal cells. Rabbit bone marrow stromal cells (MSCs) were separated from iliac crest aspirates and expanded in culture and MSCs constructed the AdBMP7. Four critical-size cranial defects were created in the rabbits with removal of osteoinductive periosteum. In 3 experimental sites, bone substitutes which were calcium metaphosphate (CMP), hydroxyapatite (HA) and HA-collagen composite (HCC) was filled. AdBMP7 was mixed in all sites. The 2D and 3D CT imaging was performed after 2, 4 and 8 weeks. After 6, 8 and 10 weeks, cranial defects sites were examined by histology study. As the results analyzed by cross-tabulation, a statistically significant difference in bone resorption and formation was noted by CT imaging at 2, 4 and 8 weeks post-bone substitutes implantation of the cranial defects, but a statistically significant difference in bone formation was not noted at 8 weeks. More bone resorption and formation on both radiological and histological observation studies was in fact observed in the CMP site and there was statistically significant. As the results analyzed by multiple regression, the time interval after implantation, the usage of bone substitutes, especially CMP, and the rabbit weight are the main factors by priority of significance. Near-complete repair of large cranial defects can be achieved using bone substitutes mixed with AdBMP7. These results showed that the use of the bone substitute as well as the integration of the stem cell concept with gene medicine is necessary and bone substitutes are capable of shortening the period of osteogenesis.

키워드

참고문헌

  1. J. M. Wozney, V. Rosen, A. J. Celeste, et al., "Novel regulators of bone formation: molecular clones and activities," Science, 242, 1528-1534 (1988).
  2. R. G. Geesink, N. H. Hoefnagels and S. K. Bulstra, "Osteogenic activity of OP-1 bone morphogenetic protein (BMP-7) in a human fibular defect," J. Bone Joint Surg., 81B, 710-718 (1999).
  3. H. Yamagiwa, N. Endo, K. Tokunaga, T. Hayami, H. Hatano and H. E. Takahashi, "In vivo bone-forming capacity of human bone marrow-derived stromal cells is stimulated by recombinant human bone morphogenetic protein-2," J. Bone Miner. Metab., 19, 0-28 (2001).
  4. J. Fang, Y. Y. Zhu, E. Smiley, et al., "Stimulation of new bone formation by direct transfer of osteogenic plasmid genes," Proc. Natl. Acad. Sci. USA, 93, 5753-5758 (1996).
  5. D. Gazit, G. Turgeman, P. Kelley, et al., "Engineered pluripotent mesenchymal cells integrate and differentiate in regenerating bone: a novel cell-mediated gene therapy," J. Gene. Med., 1, 121-133 (1999).
  6. I. Kazhdan, D. Rickard and P. S. Leboy, "HLH transcription factor activity in osteogenic cells," J. Cell Biochem., 65, 1-10 (1997).
  7. W. S. Jang, J. R. Kim, W. J. Sohn, et al., "Osteoinduction using recombinant bone morphogenetic protein-7 gene," J. of Korean Orthop. Assoc., 39, 598-606 (2004).
  8. M. C. Chang, T. Ikoma, M. Kikuchi, J. Tanaka, "The cross-linkage effect of hydroxyapatite/collagen nanocomposites on a self-organization phenomenon," J. Mater. Sci. Mater. Med.,13, 993-997 (2002).
  9. M. Kikuchi, S. Itoh, S. Ichinose, K. Shinomiya and J. Tanaka, "Self-organization mechanism in a bone-like hydroxyapatite/collagen nanocomposite synthesized in vitro and its biological reaction in vivo," Biomaterials, 22, 1705-1711 (2001).
  10. P. Chomczynski, N. Sacchi, "Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction," Anal. Biochem., 162, 156-159 (1987).
  11. A. R. Gazdag, J. M. Lane, M. Glaser and R. A. Forster, "Alternatives to autogenous bone graf: Efficacy and indication", J. Am. Acad. Orthop. Surg., 3, 1-8 (1995).
  12. C. J. Damien, J. R. Parsons, "Bone graft and bone graft substitutes:a review of current technology and applications," J. Appl. Biomater., 2, 187-208 (1991).
  13. E. Liljensten, C. Larsson, P. Thomsen, G. Blomqvist, J. M. Hirsch and C. Wedenberg, "Studies of the healing of bone grafts, and the incorporation of titanium implants in grafted bone: an experimental animal model," J. Materials Science, 9, 535-541 (1998).
  14. T. D. Alden, P. Varady, D. F. Kallmes, J. A. Jane Jr and G. A. Helm, "Bone morphogenetic protein gene therapy," Spine, 27 (16 Suppl 1), S87-S93 (2002).
  15. A. S. Breitbart, D. A. Grande, J. M. Mason, M. Barcia, R. T. James Tand Grant, "Gene-enhanced tissue engineering: Applications for bone healing using cultured periosteal cells transduced retrovirally with the BMP-7 gene," Ann. Plast. Surg., 42, 488-495 (1999).
  16. A. Minamide, M. Kawakami, H. Hashizume, T. Sakata Tamaki, "Evaluation of carriers of bone morphogenetic protein for spinal fusion," Spine, 26, 933-939 (2001).
  17. J. M. Mason, A. S. Breitbart, M. Barcia, D. Porti, R. G. Pergolizzi and D. A. Grande, "Cartilage and bone regeneration using gene enhanced tissue engineering," Clin. Orthop, 379(Suppl), S171-178 (2000).
  18. J. Y. Kim, B. Y. Kim and S. H. Kim, "A comparative study on regeneration of bone defects after the grafts of demineralized bone matrix and hydroxyapatite," Korean Journal of Anatomy, 31(1), 9-20 (1998).
  19. S. H. Kim, H. J. Kim and M. S. Kim et al., "Regeneration of artificial bone defects by allograft of demineralized bone and bone particles in rabbits," Korean Journal of Anatomy, 34(2), 193-206 (2001).
  20. L. C. Abbott, E. R. Schottstaedt, J. B. Saunders, et al., "The evaluation o cortical and cancellous bone as grafting material," J. Bone Joint. Surg., 29, 381-386 (1947).
  21. V. M. Goldberg, "Bone graft options and biologic substitutes. The biology of bone graft," Orthopedics, 26, 923-924 (2003).
  22. H. M. Lazarus, S. E. Haynesworth, S. L. Gerson, N. S. Rosenthal, A. I. Caplan, "Ex vivo expansion and subsequent infusion of human bone marrow-derived stromal progenitor cells (mesenchymal progenitor cells): Implications for therapeutic use," Bone Marrow Transplant., 16, 557 (1995).
  23. S. D. Cook, J. E. Dalton, E. H. Tan, T. S. Whitecloud, D. C. Rueger, "In vivo evaluation of recombinant human osteogenic protein implants as a bone graft substitute for spinal fusions," Spine, 19, 165524 (1994).
  24. E. A. Wang, V. D. Rosen, J. S. Alessandro, et al., "Recombinant human bone morphogenetic protein induces bone formation," Proc. Natl. Acad. Sci. USA, 87, 2220 (1990).
  25. S. C. Chang, H. Chuang, Y. R. Chen, et al., "Cranial repair using BMP-2 gene engineered bone marrow stromal cells," Surg. Res., 119(1), 85-91 (2004).
  26. T. E. Grenga, J. E. Zins, T. W. Bauer, "The role of vascularization of coralline hydroxyaptite," Plast. Reconstr. Surg., 84, 245-249 (1989).
  27. H. Schliephake, F. W. Neukam, D. Klosa, "Influence of poredimensions on bone ingrowth into porous hydroxylapatite blocks used as bone graft substitutes. A histometric study," Int. J. Oral Maxillofac. Surg., 20, 53-58 (1991).
  28. D. C. Tancred, B. A. McCormack, A. J. Carr, "A synthetic bone implant macroscopically identical to cancellous bone," Biomaterials, 19, 2303-2311 (1998).
  29. D. Y. Suh, S. D. Boden, J. Louis-Ugbo, et al., "Delivery of recombinant human bone morphogenetic protein-2 using a compression-resistant matrix in posterolateral spine fusion in the rabbit and in the non-human primate," Spine, 27, 353-360 (2002).
  30. M. Bohner, "Calcium orthophophates in medicine: from ceramics to calcium phosphate cements," Injury, 31(Suppl. 4), 37-47 (2000).
  31. J. O. Hollinger, J. Brekke, "Role of bone substitutes," Clin. Orthop., 324, 55-65 (1996).
  32. M. K. Nair, U. P. Nair, A. Seyedain, et al., "Correlation of tuned aperture computed tomography with conventional computed tomography for evaluation of osseous healing in calvarial defects," Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod., 103(2), 267-73 (2007).