Journal of the Korean Association of Oral and Maxillofacial Surgeons

  • 제30권6호
  • /
  • Pages.465-473
  • /
  • 2004
  • /
  • 2234-7550(pISSN)
  • /
  • 2234-5930(eISSN)
대한구강악안면외과학회 (The Korean Association of Oral and Maxillofacial Surgeons)
권호 표지

BDNF 유전자 이입 슈반세포와 PGA 도관을 이용한 백서 좌골신경 재생에 관한 연구

PERIPHERAL NERVE REGENERATION USING POLYGLYCOLIC ACID CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE TRANSFECTED SCHWANN CELLS IN RAT SCIATIC NERVE

  • 최원재 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 안강민 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 고은봉 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 신영민 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 김윤태 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 황순정 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 김남열 (연세대학교 의과대학 약리학교실) ;
  • 김명진 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 조승우 (한양대학교 공과대학 화공학과) ;
  • 김병수 (한양대학교 공과대학 화공학과) ;
  • 김윤희 (경희대학교 문리대학 생물학과) ;
  • 김성민 (강릉대학교 치과대학 구강악안면외과학교실) ;
  • 이종호 (서울대학교 치과대학 구강악안면외과학교실)
  • Choi, Won-Jae (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Ahn, Kang-Min (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Gao, En-Feng (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Shin, Young-Min (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Kim, Yoon-Tae (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Hwang, Soon-Jeong (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Kim, Nam-Yeol (Department of Pharmacology, Yonsei Medical School) ;
  • Kim, Myung-Jin (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Jo, Seung-Woo (Department of Chemical Engineering, Hanyang University) ;
  • Kim, Byung-Soo (Department of Chemical Engineering, Hanyang University) ;
  • Kim, Yun-Hee (Department of Biology, Kyung-Hee University) ;
  • Kim, Soung-Min (Department of Oral & Maxillofacial Surgery, College of Dentistry, Kangnung National University) ;
  • Lee, Jong-Ho (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University)
  • 발행 : 2004.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

키워드

참고문헌

  1. Schmidt CE, Leach JB: Neural tissue engineering: strategies for repair and regeneration. Annu Rev Biomed Eng 2003;5:293-347 https://doi.org/10.1146/annurev.bioeng.5.011303.120731
  2. Risitano G, Cavallaro G, Merrino T, Coppolino S, Ruggeri F: Clinical results and thoughts on sensory nerve repair by autologous vein graft in emergency hand reconstruction. Chir Main 2002;21(3):194-7 https://doi.org/10.1016/S1297-3203(02)00109-9
  3. Battiston B, Tos P, Cushway TR, Geuna S: Nerve repair by means of vein filled with muscle grafts I. Clinical results. Microsurgery 2000;20(1):32-6 https://doi.org/10.1002/(SICI)1098-2752(2000)20:1<32::AID-MICR6>3.0.CO;2-D
  4. Battiston B, Tos P, Geuna S, Giacobini-Robecchi MG, Guglielmone R: Nerve repair by means of vein filled with muscle grafts. II. Morphological analysis of regeneration. Microsurgery 2000;20(1):37-41 https://doi.org/10.1002/(SICI)1098-2752(2000)20:1<37::AID-MICR7>3.0.CO;2-5
  5. Brandt J, Dahlin LB, Lundborg G: Autologous tendons used as grafts for bridging peripheral nerve defects. J Hand Surg [Br] 1999;24(3):284-90 https://doi.org/10.1054/jhsb.1999.0074
  6. Ide C, Tohyama K, Tajima K, Endoh K, Sano K, Tamura M, et al: Long acellular nerve transplants for allogeneic grafting and the effects of basic fibroblast growth factor on the growth of regenerating axons in dogs: a preliminary report. Exp Neurol 1998;154(1):99-112 https://doi.org/10.1006/exnr.1998.6921
  7. Danielsson PA, Adolfsson L, Dahlin LB: Different effect on axonal outgrowth of application of non-absorbable or absorbable tubes around a nerve repair. Scand J Plast Reconstr Surg Hand Surg 2001;35(4):347-53 https://doi.org/10.1080/028443101317149309
  8. Vasconcelos BC, Gay-Escoda C: Facial nerve repair with expanded polytetrafluoroethylene and collagen conduits: an experimental study in the rabbit. J Oral Maxillofac Surg 2000;58(11):1257-62 https://doi.org/10.1053/joms.2000.16626
  9. Keirstead HS: Stem cell transplantation into the central nervous system and the control of differentiation. J Neurosci Res 2001;63(3):233-6 https://doi.org/10.1002/1097-4547(20010201)63:3<233::AID-JNR1016>3.0.CO;2-B
  10. Barde YA, Edgar D, Thoenen H: Purification of a new neurotrophic factor from mammalian brain. Embo J 1982;1(5):549-53
  11. Leibrock J, Lottspeich F, Hohn A, Hofer M, Hengerer B, Masiakowski P, et al: Molecular cloning and expression of brainderived neurotrophic factor. Nature 1989;341(6238):149-52 https://doi.org/10.1038/341149a0
  12. Menei P, Montero-Menei C, Whittemore SR, Bunge RP, Bunge MB: Schwann cells genetically modified to secrete human BDNF promote enhanced axonal regrowth across transected adult rat spinal cord. Eur J Neurosci 1998;10(2):607-21 https://doi.org/10.1046/j.1460-9568.1998.00071.x
  13. Utley DS, Lewin SL, Cheng ET, Verity AN, Sierra D, Terris DJ: Brainderived neurotrophic factor and collagen tubulization enhance functional recovery after peripheral nerve transection and repair. Arch Otolaryngol Head Neck Surg 1996;122(4):407-13 https://doi.org/10.1001/archotol.1996.01890160047009
  14. Shirley DM, Williams SA, Santos PM: Brain-derived neurotrophic factor and peripheral nerve regeneration: a functional evaluation. Laryngoscope 1996;106(5 Pt 1):629-32 https://doi.org/10.1097/00005537-199605000-00021
  15. Jones LL OM, Bunge MB, Tuszynski MH: Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J Physiol 2001;533:83-89 https://doi.org/10.1111/j.1469-7793.2001.0083b.x
  16. Soung-Min Kim, Jong-Ho Lee, Nam-Yeol Kim, Kang-Min Ahn, Won-Jae Choi, Si-Ho Choi et al.: Improved protocol for Schwann cell isolation and proliferation from rat dorsal root ganglia in vitro. J Kor Asso Maxillofac Plastic Reconstr Surgeons 2003;25(4):265-72
  17. Bain JR, Mackinnon SE, Hunter DA: Functional evaluation of complete sciatic, peroneal, and posterior tibial nerve lesions in the rat. Plast Reconstr Surg 1989;83(1):129-38 https://doi.org/10.1097/00006534-198901000-00024
  18. Xu X, Yee WC, Hwang PY, Yu H, Wan AC, Gao S, et al: Peripheral nerve regeneration with sustained release of poly(phosphoester) microencapsulated nerve growth factor within nerve guide conduits. Biomaterials 2003;24(13):2405-12 https://doi.org/10.1016/S0142-9612(03)00109-1
  19. Molander H, Olsson Y, Engkvist O, Bowald S, Eriksson I: Regeneration of peripheral nerve through a polyglactin tube. Muscle Nerve 1982;5(1):54-7 https://doi.org/10.1002/mus.880050110
  20. Sendtner M HB, Kolbeck R, Thoenen H, Barde YA: Brain-derived neurotrophic factor prevents the death of motorneurons in newborn rats after nerve section. Nature 1992;360:757-59 https://doi.org/10.1038/360757a0
  21. Oppenheim RW, Yin QW, Prevette D, Yan Q: Brain-derived neurotrophic factor rescues developing avian motoneurons from cell death. Nature 1992;360(6406):755-7 https://doi.org/10.1038/360755a0
  22. Yan Q, Elliott J, Snider WD: Brain-derived neurotrophic factor rescues spinal motor neurons from axotomy-induced cell death. Nature 1992;360(6406):753-5 https://doi.org/10.1038/360753a0
  23. Loh NK, Woerly S, Bunt SM, Wilton SD, Harvey AR: The regrowth of axons within tissue defects in the CNS is promoted by implanted hydrogel matrices that contain BDNF and CNTF producing fibroblasts. Exp Neurol 2001;170(1):72-84 https://doi.org/10.1006/exnr.2001.7692
  24. Li ZQ, Zhu QS, Zhu JY, Hao W, Xu YM, Wang CJ, et al: [Construction and identification of recombinant adenovirus vector containing Tet-on adjustable NT3 and BDNF genes]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2004;20(3):344-7
  25. Koda M, Hashimoto M, Murakami M, Yoshinaga K, Ikeda O, Yamazaki M, et al: Adenovirus vector-mediated in vivo gene transfer of brain-derived neurotrophic factor (BDNF) promotes rubrospinal axonal regeneration and functional recovery after complete transection of the adult rat spinal cord. J Neurotrauma 2004;21(3):329-37 https://doi.org/10.1089/089771504322972112
  26. Evans GR, Brandt K, Katz S, Chauvin P, Otto L, Bogle M, et al: Bioactive poly(L-lactic acid) conduits seeded with Schwann cells for peripheral nerve regeneration. Biomaterials 2002;23(3):841-8 https://doi.org/10.1016/S0142-9612(01)00190-9