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研究生:張朝勝
研究生(外文):Chang, Chao-sheng
論文名稱:超音波刺激結合神經導管植覆許旺細胞對神經損傷再生影響之研究
論文名稱(外文):The effects of low-intensity ultrasound on peripheral nerve regeneration in PLGA conduits seeded with Schwann cells
指導教授:徐善慧徐善慧引用關係
指導教授(外文):Hsu, Shan-hui
學位類別:碩士
校院名稱:國立中興大學
系所名稱:生命科學院碩士在職專班
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:79
中文關鍵詞:神經導管低能量超音波許旺細胞神經再生肌電圖
外文關鍵詞:Nerve guidance channelsLow-intensity ultrasoundSchwann cellNerve regenerationElectromyography
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  • 被引用被引用:2
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  • 下載下載:82
  • 收藏至我的研究室書目清單書目收藏:1
在大斷裂間隙之受損神經中,神經導管的應用以及許旺細胞 (Schwann cell) 在神經再生 (regeneration) 都扮演著極重要的角色。低能量脈波超音波已知能有效地在大白鼠神經截斷後促進其再生能力,但應用在神經導管内植覆許旺細胞之成效則是未知的。本研究將48隻雄性SD鼠的右側坐骨神經截斷成15 mm之間隙,然後分別植入不同之神經導管:(1)矽膠管+膠原蛋白 (2)聚乳酸高分子+膠原蛋白 (3)聚乳酸高分子植覆許旺細胞+膠原蛋白, 並且分別使用超音波以及假刺激 (sham stimulation) 的方式,從手術後24小時開始刺激植入之神經導管,總共接受12次超音波刺激、一天一次、每次5分鐘。分別在第四、八週進行肌電圖( EMG )檢查來分析神經再生的結果,最後在第八週利用組織切片比較各組間神經軸突 (axons) 的數目和面積,以及血管數目和肌肉重量等差異。結果發現,低能量脈波超音波刺激對於聚乳酸高分子植覆許旺細胞,確實有較好的神經修復能力,至於矽膠管則無此功效,因此利用組織工程之神經導管内植覆許旺細胞結合超音波刺激在大間隙之神經斷裂修復為可行的方法。

Low-intensity ultrasound has been proven to be effective in nerve regeneration after neurotomy. The relationship between therapeutic ultrasound and seeded Schwann cells within nerve conduits, however, is not yet known.In this study, forty-eight male Spraque-Dawley rats , weighing 250-300g were used. The right sciatic nerves of the rates were transected with gaps of 15mm and bridged with nerve conduits . Three control groups were used : silicone+collagen, PLGA+collagen and PLGA with Schwann cells+collagen. All conduits were received 12 ultrasound treatment sessions over 2 weeks after 24h neurotomy. Ultrasound was applied as follows: frequency 1MHz, intensity 0 W/cm2 (sham) or 0.3W/cm2 (SATP), treatment 5 mins/day. The sham groups were utilized under the same condition for comparison. Electromyography was assessed by the motor unit action potential (MUAP) and fibrillation potential (Fib) measured at 4 and 8 weeks during all periods. The specimens of conduits were harvested for histological analysis at 8 weeks.The results indicated that PLGA + Schwann cells( 9000 cells )with pulsed wave ultrasonic stimulation resulted in a significant acceleration in number of regenerated axons at the midconduit portion of the implanted grafts, as compared to sham groups. We also found that ultrasonic stimulation on silicone groups induced mass fibrous tissues that covered the nerve conduits and retarded the axon regeneration. It was suggested that ultrasonic stimulation be a new and effective technique for nerve regeneration on Schwann cells-seeded PLGA conduits. Nevertheless, the application of ultrasound may not be suitable for silicone conduits in nerve regeneration.

目錄 頁次
目錄 . i
圖目錄 iv
表目錄 vi
第一章 緒論 1
1-1 前言 1
1-2 研究目的 2
第二章 文獻回顧 3
2-1 周圍神經系統 3
2-1-1 周圍神經的構造 3
2-1-2 周圍神經損傷的分類 4
2-1-3 周圍神經損傷的病理變化 6
2-1-4 周圍神經損傷的再生 8
2-2 組織工程神經修復導管 9
2-2-1 非生物降解神經導管 9
2-2-2 生物降解神經導管 10
2-2-3 神經導管的結構與物理特性 10
2-2-4 許旺細胞植覆神經 11
2-2-5 神經營養因子 12
2-2-6 未來的展望 12
2-3 超音波物理刺激 13
2-3-1 超音波概述 13
2-3-2 超音波的物理特性 13
2-3-3 超音波的生物效應 15
2-3-4 超音波的臨床應用 18
2-4 神經電氣生理檢查 20
2-4-1 神經傳導檢查( Nerve Conduction Velocity, NCV ) 21
2-4-2 肌電圖檢查( Electromyography, EMG ) 21
第三章 材料與方法 23
3-1 實驗材料 23
3-1-1 聚乳酸-聚甘醇酸聚合物( PLGA )神經導管的製作 23
3-1-2 神經細胞的取得與純化 23
3-1-3 PLGA導管植覆許旺細胞 24
3-1-4 超音波刺激器 24
3-2 實驗動物模式 25
3-2-1 動物植入實驗 25
3-2-2 實驗動物飼養環境 26
3-2-3 超音波刺激 26
3-2-4 肌電圖檢查 27
3-2-5 神經導管及神經再生情形 27
3-2-6 組織學定性分析 28
3-2-7 組織學定量分析 28
3-2-8 統計分析 29
第四章 結果 30
4-1 大白鼠外觀 30
4-2 神經導管外觀 30
4-3 神經導管成功修復比例 31
4-4 肌電圖檢查分析 32
4-4-1 顫動電位( fibrillation, Fib ) 32
4-4-2 運動單位電位( motor unit action potential, MUAP ) 38
4-5 神經再生組織學分析 42
4-5-1 神經軸突數目 42
4-5-2 軸突面積 44
4-5-3 再生血管數目 45
4-6 腓腸肌重量比例分析 46
第五章 討論 48
第六章 結論 56
參考文獻 57
圖目錄 頁次
圖 1 、 神經元的基本構造 63
圖 2 、 周圍神經的結締組織 63
圖 3-1、第八週實驗動物植入之神經導管外觀 64
圖 3-2、第八週從實驗動物Group 2取下之神經導管外觀 64
圖 3-3、第八週從實驗動物取出之PLGA神經導管外觀 65
圖 4 、 各組再生神經成功長過 15 mm 間隙的比例 65
圖 5 、 第四週神經成功再生的顫動電位( Fib )波型 66
圖 5-1、 第四週神經成功再生的顫動電位( Fib ) 66
圖 5-2、 第八週神經成功再生的顫動電位( Fib ) 波型 67
圖 5-3、 第八週神經成功再生的顫動電位( Fib ) 67
圖 5-4、 第四週和第八週神經成功再生 Fib 於各組間的變化 68
圖 5-5、 第四週無神經再生的顫動電位( Fib ) 68
圖 5-6、 第八週無神經再生的顫動電位( Fib ) 69
圖 6 、 第四週神經成功再生的運動單位電位( MUAP )波型 69
圖 6-1、 第四週神經成功再生的運動單位電位( MUAP ) 70
圖 6-2、 第八週神經成功再生的運動單位電位( MUAP )波型 70
圖 6-3、 第八週神經成功再生的運動單位電位( MUAP ) 71
圖 6-4、 第四週和第八週神經成功再生MUAP 於各組間的變化 71
圖 7-1 ~ 圖7-6、各組再生神經中段之神經橫切面圖 72
圖 7-1、Group 1 . Silicone tube/collagen, US(-) 72
圖 7-2、Group 2 . Silicone tube/collagen, US(+) 72
圖 7-3、Group 3 . PLGA tube/collagen, US(-) 73
圖 7-4、Group 4 . PLGA tube/collagen, US(+) 73
圖 7-5、Group 5 . PLGA tube/cell/collagen, US(-) 74
圖 7-6、Group 6 . PLGA tube/cell/collagen, US(+) 74
圖 8 、 第八週各組神經中段再生神經橫切面的軸突數目 75
圖 9 、 第八週各組神經中段再生神經橫切面的軸突面積 75
圖 10 、第八週各組神經中段再生神經橫切面的再生血管數目 76
圖 11 、第八週各組腓腸肌重量比例分析 76
圖 11-1、正常腓腸肌的組織橫切面圖 77
圖 11-2、第八週PLGA導管組腓腸肌的組織橫切面圖 77
表目錄 頁次
表1、 六個動物實驗組別及超音波刺激模式 78
表2、 六個動物實驗組神經成功再生的Fib結果分析 78
表3、 六個動物實驗組無神經再生的Fib結果分析 79
表4、 六個動物實驗組神經成功再生的MUAP結果分析 79

參考文獻
〔1〕陳俊男,神經修復導管發展之現況,化工資訊,2002年12月,40-45.
〔2〕Evans GRD, Brandt K, Widmer MS, Lu L, Meszlenyi RK. In vivo evaluation of poly(L-lactic acid) porous conduits for peripheral nerve regeneration. Biomaterials 1999;20:1109-1115.
〔3〕Waid HL, Sarakinos G, Lyman MD, Mikos AG, Vacanti JP, and Langer R. Cell Seeding in Porous Transplantation Devices. Biomaterials 1993;14:270-278.
〔4〕Crisci AR, Ferreira AL. Low-intensity pulsed ultrasound accelerates the regeneration of the sciatic nerve after neurotomy in rats.
Ultrasound Med Biol 2002;28:1335-1341.
〔5〕王宗道編譯,巴氏神經解剖學,藝軒圖書出版社,1992.
〔6〕Griffin JW, Kidd G, Trapp BD. Interactions between axons and schwann cells. In Dyck PJ, Thomas PK, eds. Peripheral neuropathy. Phialdelphia: WB Sanders Co, 1993.
〔7〕Tonge DA, Golding JP. Regeneration and repair of the peripheral nervous system. Sem Neurosci 1993;5:385-390.
〔8〕Seddon HJ. Surgical disorders of the peripheral nerves. Baltimore: Williams and Wilkins, 1972.
〔9〕Sunderland S. Nerves and nerve injuries. New York: Churchill Livingston,1981.
〔10〕牛建昭主編,組織學與胚胎學,人民衛生出版社,2002.
〔11〕Fawcett JW, Keynes RJ. Peripheral nerve regeneration. Annu Rev neurosci 1990;13:43-60.
〔12〕Lundborg G. Nerve regeneration and repair. Acta Orthop Scand 1987;58: 145-169.
〔13〕Lanza RP, Langer R, Vacanti J. Principles of tissue engineering. Academic press. 2000;56:785-798.
〔14〕Terzis JK, Sun DD, Thanos PK. Historical and basic science review: past, present and future of nerve repaired. J Reconstr Microsurg 1997;13:215-225.
〔15〕Millesi H. Progress in peripheral nerve reconstruction. World J Surg 1990;14:733-747.
〔16〕Aldini NN, Perego G, Cella GD. Effectiveness of a bioabsorbable conduit in the repair of peripheral nerves. Biomaterials 1996;17:959-962.
〔17〕Archibald SJ, C Krarup, J Shefner, ST Li and RD Madison. A collagen-based nerve guide conduit for peripheral nerve repair: An electrophysiological study of nerve regeneration in rodents and nonhuman primates. J Comp Neurol 1991;306:685-696.
〔18〕Robinson PH, Lei BV, HJ Hoppen, JW Leenslag, AJ Pennings and P Nieuwenhuis. Nerve regeneration through a two-poly biodegradable nerve guide in the rat and the influence of ACTH
4 -9 nerve growth factor. Microsurgery 1991;12:412-419.
〔19〕Tountas CP, RA Bergman, TW Lewis, HE Stone, JD Pyrek, and HV Mendenhall. A comparison of peripheral nerve repair using an absorbable tubulization device and conventional suture in primates. J Appl Biomater 1993;4:261-268.
〔20〕Pappas, Langer RS. Biopolymer II. Springer,1995.
〔21〕Schugens CH, V Maquet, Ch Grandfils, R Jerome . Polylactide macroporous biodegradable implants for cell transplantation. II. Preparation of polylactide foams by liquid-liquid phase sepration. J Biomed Mater Res 1996;30:449-461.
〔22〕Lundborg G. Nerve regeneration: the nerve chamber as an experimental tool. In : Nerve injury and repair. New York: Churchill Livingstone, 1998.
〔23〕黃義侑,生分解性PLA及其共聚物在醫學上的應用,化工技術,1996; 4(7):152-159.
〔24〕張耀南,漫談乳酸聚合物在生物醫學材料的應用與發展,化工技術, 1996;4(7):174-178.
〔25〕Yoshii S, Oka M, Shima M, Taniguchi A, Akagi M. 30 mm regeneration of rat sciatic nerve along collagen filaments. Brain Res 2002;949:202-208.
〔26〕Mickinnon SE, Dellon AL. Clinical nerve reconstruction with a bioabsorbable polyglycolic acid tube. Plast Reconstr Surg 1990;85: 419-424.
〔27〕Yin Q, Kemp GJ, Frostick SP. Neurotrophins, neurons and peripheral nerve regeneration. J of Hand Surgery 1998;23:433-437.
〔28〕Fine EG, Decosterd I, Papaloizos M, Zurn AD, Aebischer P. GDNF and NGF released by synthetic guidance channels support sciatic nerve regeneration across a long gap. Eur J Neurosci 2002;15:589-601.
〔29〕Heath CA, Rutkowski GE. The development of bioartificial nerve grafts for peripheral nerve regeneration. Tibtech 1998;16:163-168.
〔30〕Tessa H, Cathryn S, Daniel H, Mack C, Joseph P. A polymer foam conduit seeded with Schwann cells promotes guided peripheral nerve regeneration. Tissue Eng 2000;6:119-127.
〔31〕Gregory RD, Keith B, Steven K, Priscilla C, Lisa O. Bioactive poly (l-lactic acid) conduits seeded with Schwann cells for peripheral nerve regeneration. Biomaterials 2002;23:841-848.
〔32〕Douglas WZ. The microenvironment of injured and regenerating peripheral nerves. Muscle Nerve Suppl 2000;9:S33-S38.
〔33〕Levi AD, Guenard V, Aebischer P, Bunge RP. The function characteristics of Schwann cells cultured from human peripheral nerve after transplantation into a gap within the rat sciatic nerve. J Neurosci 1994b;14:1309-1319.
〔34〕Terenghi G. Peripheral nerves injury and regeneration . Histol Histopathol 1995;10:709-718.
〔35〕Chen ZW, Wang MS. Effects of growth factor on crushed sciatic nerve regeneration in rats. Microsurgery 1995;16:547-551.
〔36〕Derby A, Engleman VW, Gregory E. Frierdich . Nerve growth factor facilitates regeneration across nerve gap: Morphological and behavioral studies in rat sciatic nerve. Experi Neuro 1993;119:176-191.
〔37〕Chen YS, Hsieh CL, Tsai CC . Peripheral nerve regeneration using silicone rubber chambers filled with collagen, laminin and fibronectin. Biomaterials 2000;21:1541-1547.
〔38〕Verdu E, Labrador RO, Rodriguez FJ. Alignment of collagen and laminin-containing gels improve nerve regeneration within silicone tubes Restor Neurol Neurosci 2002;20:169-179.
〔39〕Wells PNT. Biomedical ultrasonics. London: Academic Press,1977.
〔40〕Michlovitz SL. Thermal agents in rehabilitation. Philadelphia: McGraw and Hill,1986.
〔41〕Dyson M, Brookrs M. Stimulation of bone repair by ultrasound. Ultrasound Med Biol 1983;9(Suppl. 2):61-66.
〔42〕Heckman JD, Ryaby JP, McCabe J . Acceleration of tibial fracture -healing by non-invasive , low intensity pulsed ultrasound. J Bone Joint Surg 1994;76:26-34.
〔43〕Tsai CL, Chang WH, Liu TK. Preliminary studies of duration of intensity of ultrasonic treatment on fracture repair. Chin J Physiol 1992;35:21-26.
〔44〕Enwemeka CS, Rodriguez O, Mendosa S. The biomechanical effects of low-intensity ultrasound on healing tendond. Ultrasound Med Biol 1990;16:801-807.
〔45〕Hogan RDB, Burke KM, Franklin TD. The effect of ultrasound on the microvascular hemodynamics in skeletal muscle: Effects during ischemia. Microvasc Res 1982;23:370-379.
〔46〕Bierman W. Ultrasound in the treatment of scars. Arch Phys Med Rehabil 1976;35:209-214.
〔47〕Dyson M. Role of ulttrasound in wound healing. In: Kloth LC, McCulloch JM, Feedar JA, eds. Wound healing: Alternatives in management. Philadelphia: FA Davis, 1990.
〔48〕Moll MJ. A new approach to pain : Lidocaine and decadron with ultrasound. Phys Ther 1977;54:594-598.
〔49〕吳進安,神經診斷學,國立編譯館,1996.
〔50〕David CP, Barbars ES. Electromyography and neuromuscular disorders. Boston :Butterworth and Heinemann,1998.
〔51〕Chen YS, Liu CJ, Cheng CY. Effect of bilobalide on peripheral nerve regeneration. Biomaterials 2004;25:509-514.
〔52〕Seckel BR, Chiu TH, Nyilas E . Nerve regeneration through synthetic biodegradable nerve guides: regulation by the target organ. Plast Reconstr Surg 1984;74:173-181.
〔53〕Moore JH, Gieck JH, Saliba EN et al. The biophysical effects of ultrasound on median nerve distal latencies. Electromyogr Clin Neurophysiol 2000;40: 169-180.
〔54〕Carstensen EL. Biological effects of acoustic cavitation. First symposium on safety and standardisation of ultrasound in obstetrics. Ultrasound Med Biol 1986;12:703-704.
〔55〕Holland C, Apfel R. Threshold of transient cavitation produced by pulsed ultrasound in a controlled nuclei envirommen. J Acoust Soc Am 1990;88: 2059-2069.
〔56〕曹涌,成紅兵,王曉冬. 殼聚糖管與聚乙醇酸織維複合移植修復大鼠坐 骨神經缺損研究. 中華顯微外科雜誌, 2002;25(4): 274-277.
〔57〕Kraft GH. Fibrillation potential amplitude and muscle atrophy following peripheral nerve injury. Muscle Nerve 1990;13:814- 821.
〔58〕Stalberg EV, Antoni L. Electrophysiological cross section of the motor unit. J Neurol Neurosurg Psychiatry 1980;43:469-474.
〔59〕Chang CJ, SH Hsu. The effects of low-intensity ultrasound on peripheral nerve regeneration in poly(DL-lactic acid-co-glycolic acid) conduits seeded with Schwann cells . Ultrasound Med Biol (in press).

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