Journal of Biomedical Engineering Research (대한의용생체공학회:의공학회지)

The Korean Society of Medical and Biological Engineering (대한의용생체공학회)
권호 표지
DOI QR코드

DOI QR Code

Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture

기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증

  • Lee, Yong-Jae (Department of Biomedical Engineering, Inje University) ;
  • Shin, Ji-Won (Department of Biomedical Engineering, Inje University) ;
  • Shin, Ho-Jun (School of Material Science, Japan Advanced Institute of Science and Technology) ;
  • Kim, Chan-Hwan (Department of Pathology, Inje University) ;
  • Park, Ki-Dong (Department of Molecular Science and Technology, Ajou University) ;
  • Bae, Jin-Woo (Department of Molecular Science and Technology, Ajou University) ;
  • Seo, Hyoung-Yeon (Department of Orthopaedic Surgery, College of Medicine, Chunnam National University) ;
  • Kim, Young-Jick (Department of Dental Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Shin, Jung-Woog (Department of Biomedical Engineering, Inje University)
  • 이용재 (인제대학교 의용공학과) ;
  • 신지원 (인제대학교 의용공학과) ;
  • 신호준 (일본 자이스트 재료과학과) ;
  • 김찬환 (인제대학교 병리학과 병리학교실) ;
  • 박기동 (아주대학교 분자과학기술학과) ;
  • 배진우 (아주대학교 분자과학기술학과) ;
  • 서형연 (전남대학교 의과대학 정형외과) ;
  • 김영직 (부산가톨릭대학교 치기공과) ;
  • 신정욱 (인제대학교 의용공학과, 인제대학교 FIRST 연구그룹, 의생명공학원)
  • Published : 2007.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

Keywords

References

  1. J. S. Ahn, J. K. Lee, T. S. Jeon, S. Y. Kwon and S. K. Kwak, 'Correlation among magnetic resonance images, electron microscopic finding, light microscopic findings and clinical symptom of the degeneration of lumbar intervertebral disc,' J. Korean Soc. Spine Surg., vol. 8, no. 2. pp. 121-129, 2001 https://doi.org/10.4184/jkss.2001.8.2.121
  2. B. Shen, J. Melrose, P. Ghosh and F. Tayler, 'Induction of matrix metalloproteinase-2 and -3 activity in ovine nucleus pulposus cells grown in theree-dimensional agarose gel culture by interleukin-$1{\beta}$: a potential pathway o disc degeneration,' Eur. Spine J., vol. 12, pp. 66-75, 2003
  3. J. A. Buckwalter, T. A. Einhorn and S. R. Simon, 'Orthopaedic basic sicence 2nd. American academy of orthopaedic surgeons,' Illinois, pp. 443, 2000
  4. J. C. Gan, P. Ducheyne, E. J. Vresilovic, W. Swaim and I. M. Shapiro, 'Intervertebral disc tissue engineering I : Characterization of the nucleus pulposus,' Clinical Orthop Related Res, vol. 411, pp. 305-314, 2003 https://doi.org/10.1097/01.blo.0000063796.98363.9a
  5. H. S. An, E. J. Thonar and K. Masuda, 'Biological repair of intervertebral disc,' Spine, vol. 28, no. 15S, pp. S86-S92, 2003 https://doi.org/10.1097/00007632-200308011-00015
  6. H. E. Gruber, G. L. Hoeslscher, K. Leslie, J. A. Ingram and E. N. Hanley, 'Three-dimensional culture of human disc cells within agarose or a collagen sponge: assessment of proteoglycan production,' Biomaterials, vol. 27, pp. 371-376, 2006 https://doi.org/10.1016/j.biomaterials.2005.06.032
  7. R. Q. Brown, A. Mount and K. J. L. Burg, 'Evaluation of polymer scaffolds to be used in a composite injectable system for intervertebral disc tissue engineering,' J. Biomed. Mater. Res. A, vol. 74, no. 1, pp. 32-39, 2005
  8. T. Chujo, H. S. An, K. Akeda, K. Miyamoto, C. Muehleman, M. Attawia, G. Andersson and K. Masuda, 'Effects of growth differentiation of factor-5 on the intervertebral disc - In vitro bovine study and in vivo rabbit disc degeneration model study,' Spine, vol. 31, no. 25, pp. 2909-2917, 2006 https://doi.org/10.1097/01.brs.0000248428.22823.86
  9. S. J. Chacko, J. Abbott S. and Holtzer, 'The loss of phenotypic traits by differentiated cells. VI. Behavior of the progeny of a single chondrocyte,' J. Exp. Med., vol. 130, pp. 417-442, 1969 https://doi.org/10.1084/jem.130.2.417
  10. M. V. Risbud, M. W. Izzo, C. S. Adams and et al. 'An organ culture system for the study of the nucleus pulposus: description of the system and evaluation of the cells,' Spine, vol. 28, no. 24, pp. 2652-2659, 2003 https://doi.org/10.1097/01.BRS.0000099384.58981.C6
  11. C. R. Lee, J. C. Latridis, L. Poveda and M. Alini, 'In vitro organ culture of the bovine intervertebral disc - Effects of vertebral endplate and potential for mechanobiology studies,' Spine, vol. 31,no. 5,pp. 515-522, 2006 https://doi.org/10.1097/01.brs.0000201302.59050.72
  12. S. Stern, K. Lindenhayn, O. Schultz and C. Perka, 'Cultivation of porcine cells from the nucleus pulposus in a fibrin/hyaluronic acid matrix,' Acta Orthop Scand, vol. 71, no. 5, pp. 496-502, 2000 https://doi.org/10.1080/000164700317381207
  13. J. W. Lee, M. C. Jung, H. D. Park, K. D. Park and G. H. Ryu, 'Synthesis and characterization of thermosensitive chitosan copolymer as a novel biomaterial,' J. Biomater Sci. Polym. Ed., vol. 15, no. 8, pp. 1065 - 1079, 2004 https://doi.org/10.1163/1568562041526496
  14. M. Okuma, J. Mochida, K. Nishimura, K. Sakabe and K. Seiki, 'Reinsertion of stimulated nucleus pulposus cells retards intervertebral disc degeneration: an in vitro and in vivo experimental study,' J. Orthop. Res., vol. 18, pp. 988 - 997, 2000 https://doi.org/10.1002/jor.1100180620
  15. G. D. Palmer, Ph P. H. Chao, F. Raia, R. L. Mauck, W. B. Valhmu and C. T. Hung, 'Time-dependent aggrecan gene expression of articular chondrocytes in reponse to hyperosmotic loading,' Osteoarthritis cartilage, vol. 9, no. 8, pp. 761-770, 2001 https://doi.org/10.1053/joca.2001.0473
  16. H. Ishihara, K. Warensjo, S. Roberts and J. P.Urban, 'Proteoglycan synthesis in the intervertebral disk nucleus: the role of extracellular osmolaity,' Am. J. Physiol., vol. 272, pp. 1499-1506, 1997 https://doi.org/10.1152/ajpcell.1997.272.5.C1499
  17. W. Johannessen, E. J. Vresilovic, A. C. Wright, D. M. Elliott, 'Intervertebral disc mechanics are restrored following cyclic loading and unloaded recover,' Annals of Biomedical Engineering, vol. 32, no. 1, pp. 70-76, 2004 https://doi.org/10.1023/B:ABME.0000007792.19071.8c
  18. C. Park, Y. J. Kim, C. S. Lee, K. An, H. J. Shin, C. H. Lee, C. H. Kim and J. W. Shin, 'An in vitro animal study of the biomechanical responses of anulus fibrosus with aging,' Spine, vol. 30, no. 10, pp. E259-265, 2005 https://doi.org/10.1097/01.brs.0000162531.49297.43
  19. D. Haschtmann, J. V. Stoyanov, L. Ettinger, L. P. Nolte and S. J. Ferguson, 'Establishment of a novel intervertebral disc/endplate culture model: Analysis of an ex vivo in vitro whole-organ rabbit culture system,' Spine, vol. 31, no. 25, pp. 2918-2925, 2006 https://doi.org/10.1097/01.brs.0000247954.69438.ae
  20. J. C. Lotz, 'Animal models of intervertebral disc degeneration: Lessons learned,' Spine, vol. 24, no. 23, pp. 2742-2750, 2004
  21. K. Takegami, H. S. An, F. Kumano, K. Chiba, E. J. Thonar, K. Singh and K. Masuda, 'Osteogenic protein-I is most effective in stimulating nucleus pu1posus and annulus fibrosus cells to repair their matrix after chondroitinase ABC-induced in vitro chemonucleolysis,' Spine J, vol. 5, no. 3, pp. 231-238, 2005 https://doi.org/10.1016/j.spinee.2004.11.001
  22. S. Sobajima, J. F. Kompel, J. S. Kim, C. J. Wallach, D. D. Robertson, M. T. Vogt. J. D. Kangand L. G. Gilbertson, 'A slowly progressive and reproducible animal model of intervertebral disc degeneration characterized by MRI, X-ray, and histology,' Spine, vol. 30, no. 1, pp. 15-24, 2005 https://doi.org/10.1097/01.brs.0000148048.15348.9b
  23. Y. S. Park, J. L. Cho and J. P. Kostuik, 'Experimental annulotomy-induced degeneration in rabbit intervertebral discs -Comparative study between incomplete and complete annulotomy,' J. of Korean Orthop Accoc, vol. 38, pp. 619-623, 2003 https://doi.org/10.4055/jkoa.2003.38.6.619
  24. Y. T. Kim, J. J. Kim, J. H. Hwang, M. J. Shin, E. S. Yu and Y. K. Shin, 'Annulus tears in the intervertebral disc degeneration-An experimental study using an animal model,' J. of Korean Spine Surg, vol. 1, pp. 259-272, 1994