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The application of nerve conduit in peripheral nerve reconstruction
|關鍵字:||microgrooves;微溝槽;asymmetric conduits;polylactide (PLA);chitosan;fibroblast growth factor;plasma surface modification;neural stem cells;nerve regeneration.;非對稱導管;聚乳酸;幾丁聚醣;纖維母細胞生長因子;電漿表面處理;神經幹細胞;神經再生||出版社:||化學工程學系所||引用:||1. Mooney DJ, Baldwin DF, Suh NP, Vacanti JP, Langer R. Novel approach to fabricate porous sponges of poly(D,L-lactic-co-glycolic acid) without the use of organic solvents. Biomaterials 1996; 17: 1417-1422. 2. Gray JE, Norton PR, Griffiths K. Surface modification of a biomedical poly(ether)urethane by a remote air plasma. Appl Surf Sci 2003; 217: 210-222. 3. Dai WS, Barbari TA. Hollow fiber-supported hydrogels with mesh-size asymmetry. J Memb Sci 2000; 171: 79-86. 4. Elbert DL, Herbert CB, Hubbell JA. Thin polymer layers formed by polyelectrolyte multilayer techniques on biological surfaces. Langmuir 1999;15: 5355-5362. 5. Ho MH, Hou LT, Tu CY, Hsieh HJ, Lai JY, Chen WJ, et al. 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Webb K, Hlady V, Tresco PA. Relative importance of surface wettability and charged functional groups on NIH 3T3 fibroblast attachment, spreading, and cytoskeletal organization. J Biomed Mater Res 1998; 41: 422-430. 39. Shelton RM, RasmussenAC, Davies JE. Protein adsorptionat the interface between charged polymer substrata and migrating osteoblasts. Biomaterials 1988; 9: 24-29. 40. Tahara M, Cvong NK, Nakashima Y. Improvement in adhesion of polyethylene by glow-discharge plasma. Surf Coat Technol 2003; 174: 826-830. 41. Greene G, Yao G, Tannenbaum R. Wetting characteristics of plasma-modified porous polyethylene. Langmuir 2003;19: 5869-5874. 42. Sartori S, Rechichi A, Vozzi G, D’Acunto M, Heine E, Giusti P et al. Surface modification of a synthetic polyurethane by plasma glow discharge: Preparation and characterization of bioactive monolayers. React. Funct. Polym. 2008; 68: 809-821. 43. Pataky DM, Borisoff JF, Fernandes KJ, Tetzlaff W, Steeves JD. Fibroblast growth factor treatment produces differential effects on survival and neurite outgrowth from identified bulbospinal neurons in vitro. Exp. Neurol. 2000; 163: 357-372. 44. Cordeiro PG, Seckel BR, Lipton SA, D’Amore PA, Wagner J, Madison R, Acidic fibroblast growth factor enhances peripheral nerve regeneration in vivo. Plast Reconstr Surg 1989; 83: 1013-1019. 45. Lee LM, Huang MC, Chuang TY, Lee LS, Cheng H, Lee IH. Acidic FGF enhances functional regeneration of adult dorsal roots. Life Sci 2004; 74: 1937-1943. 46. Sakiyama-Elbert SE, Hubbell JA. Development of fibrin derivatives for controlled release of heparin-binding growth factors. J Control Release 2000; 65: 389-402. 47. Midha R, Munro CA, Dalton PD, Tator CH, Shoichet MS. Growth factor enhancement of peripheral nerve regeneration through a novel synthetic hydrogel tube. J Neurosurg. 2003; 99: 555-565. 48. 林哲永，大氣電漿改質神經導管材料於周邊神經之應用。國立中興大學碩士論文 民96||摘要:||
在論文第一部分中，利用一結合相轉移法與微轉印法於同一步驟完成的新穎技術製作一神經導管應用於周邊神經的修復。此導管同時具有可提供物質非對稱流通性的特殊非對稱孔洞及導引細胞排序的表面溝槽。在修復10 mm坐骨神經缺陷的大鼠動物實驗中，與其他實驗組相比，同時具有非對稱及微溝槽這兩種結構的聚乳酸導管在4週有最佳的髓鞘化情形且於6週有最多的血管生成。動物步行分析結果中也顯示此特殊結構的神經導管提供較佳的功能性恢復。在論文第二部分中為進一步提昇修復效果，含奈米金的幾丁聚醣與纖維母細胞生長因子以大氣電漿依序接枝於此導管表面，並以表面化學分析作確認。此改質的導管確實於10 mm坐骨神經缺陷的大鼠動物實驗中增進了神經的修復。在第三部分中，證明了接枝於含奈米金-幾丁聚醣表面的纖維母細胞生長因子在釋放後才能保持較佳的活性。進一步以大間隙（15mm）大鼠坐骨神經缺陷實驗作為驗證，此一導管在修復程度及功能性恢復上確有較佳的表現。若進一步植入神經幹細胞於導管內，在12週後再生神經的髓鞘化程度及再生面積都發現更進一步的提升。於6週時，複合動作電位的波形與自體移植的控制組非常相似；另神經傳導速率在12週可達自體移植組的九成。植入6週後，在接枝有奈米金-幾丁聚醣與纖維母細胞生長因子的導管內，存活的部分神經幹細胞分化為似神經膠細胞，並於再生神經中發現較多的神經幹細胞。結合良好設計的基材、生長因子與幹細胞這三大要素的組織工程神經導管在未來或有可能修復嚴重受損的神經。
In the first part, an innovative technique combining phase transition and microprinting in one step was applied to fabricate the nerve conduits used in peripheral nerve regeneration. The asymmetric microporosity served to generate asymmetric permeability and the surface microgrooves were introduced to achieve cell alignment in vitro. The symmetric/asymmetric porous poly(D,L-lactide) (PLA) substrates with microgrooves on the surface were tested for their ability to repair 10 mm sciatic nerve transection defects in rats. The in vivo results showed that the regenerated nerves in the asymmetric conduits with surface microgrooves had highest degree of myelination at 4 weeks and the most number of vessels at 6 weeks. The walking track analysis also implied that the asymmetric conduits with surface microgrooves had the highest degree of functional recovery. To further improve the performance, chitosan (containing nano gold) and fibroblast growth factor 1 (FGF1) were sequentially grafted on the microgrooved PLA surface by the assistance of open air plasma treatment in the second part. Grafting of these components was verified with electron spectroscopy for chemical analysis (ESCA). The modified nerve conduits showed enhanced ability in the repair of 10 mm sciatic nerve transection defects in rats. In the third part, grafting of FGF1 was required to be performed on chitosan-nano Au modified surface for better activity of the released FGF1. The performance of the PLA conduits grafted by chitosan-nano Au and FGF1 also had the best result in the regeneration capacity and in promoting the functional recovery of sciatic nerve with large defect (15 mm) in rats. If the conduits were seeded with neural stem cells (NSCs), the degree of myelination and the area of regenerated nerve were further enhanced after 12 weeks. This was evident by the waveform of compound muscle action potentials which was relatively similar to that in autografts at 6 weeks, as well as the nerve conduction velocity which achieved about 90% of that in autografts at 12 weeks. Living NSCs were demonstrated in the regenerated nerve tissue after 6 weeks of implantation. Some NSCs had partially differentiated into glia-like cells. More NSCs were found in the regenerated nerve of the conduits if the nerve conduit employed had better performance prior to cell seeding. It is thus possible to consolidate tissue engineering nerve conduits with well-designed substrate, growth factors and stem cells that synergistically contribute to regenerate the severely damaged nerve.
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