201224149 六、發明說明： 【發明所屬之技術領域】 [0001] 本發明涉及一種神經移植體，尤其係涉及一種可供生物 體移植的神經移植體。 [先前技術] [喔]神經系統主要由神經元（n e U r ο n s )以及神經膠質細胞 (neuron glia 1 cells)構成一複雜且特異的溝通網域 ，用以與其他組織或器官建立連結以進行功能協調。神 〇201224149 VI. Description of the Invention: [Technical Field] [0001] The present invention relates to a nerve graft, and more particularly to a nerve graft for transplantation of a living body. [Prior Art] [喔] The nervous system consists mainly of neurons (ne U r ο ns ) and neuron glia 1 cells, which form a complex and specific communication domain for establishing links with other tissues or organs. Perform functional coordination. God
[0003] 099143857 經系統中，係由神經元來執疗接收刺激、通過傳導並輸 出神經遞質（neuron _tr.ansmi tter)以進行組織或器官 間訊息溝通’而神經膠質細胞則執行神經尽物理性支援 、營養提供以及調節溝通訊基速度等功能。每一神經元 依據型態包含胞體（cell body)與神經突起（neurite) 兩部分’神經突起自胞體延伸並朝向其他神經元或係其 他細胞（例如：肌肉細胞）生長，其中神經突起又分為軸 突Ux〇n)與樹突（dendrite)兩耧。一般來說’刺激由 樹大接收並將衝動傳向胞鐘，衝動經過轴突傳導至轴突 末端，並微料物質來_其他細胞。 由於神經系統扮演生物體内各組織與器官之間的協調作 用，其重要性不言可令。先前，因神經系統受損而導致 的神經缺損係臨騎㈣致雜疾病。其，中，通過植入 H雜體來修復受損的神經系統，係神經外科手術用 來修錢各種情㈣起㈣㈣統損錢—種重要仔 。先前的神經移植體通常為 位兩端的神經管在神、㈣統受損部 該神”由生物降解材料製成的管狀 .表單編號A0101 第3頁/共40頁 0992075912-0 201224149 [0004] [0005] [0006] [0007] 099143857 結構。神經系統受損部位1的㈣“_神經管内 ㈣Μ㈣”經系統受損部位的另一 端。 通常，需要通過植人所述神㈣的方式進行修復的的受 損部位的長度較長，而所心申經_常緩 f又故洲所述神經官來修復受損的神經系統所需的 修復時間較長。 【發明内容】 有審於此，提供—種減少__經系統的修復時間神 1 經移植體的製備方法實為必秦。 -種神經移植體，其包括―疏水性基底、―奈米礙管膜 結構、一蛋白質層及—神經網路》所述奈米碳管膜結構 設置在所述疏水性基底的―表^^所述蛋白質層設置在 所述奈米碳管赌構_所魏水絲底的表面。所述 神經網路設置在所述蛋白質層遠離所述奈米碳管膜結構 的表面力述神經網路包括多個神經細胞及多個神經突 起’所述多個神經細胞之間的神經突起相互連接形成一 ί 神經網路。 相較於麟技術’所料經移植體在所述培育層的表面 形成所述神經網路。所述培育層中的奈米碳管膜結構與 疏水性基底均具有彈性佳與延展性良好等優點故，所 述神經移植體可根據受損神經系統的受損部位的形狀、 大小進行裁剪、拉伸並楂人受損部位。所述神經網路具 有生物活性及信號傳遞能力，從而使得包括所述神經網 路的神經移植體也具有生物活性及信號傳遞能力。當所 表早編號Α0101 第4頁/共40頁 0992075912-0 201224149 述神經移植體植入生物體中的受損部位時，由於所述神 經植入體中的神經元與所述受損部位兩端或邊緣的神經 元的距離非常近，故可通過直接縫合所述神經植入體中 的神經元與受損部位邊緣的神經元的方式使所述受損部 位的兩端建立起信號傳遞能力，完成受損部位的神經修 復，從而節省所述神經突起的生長時間，減少受損的神 經系統的修復時間。 【實施方式】 _ [0008] 請參閱圖1，本發明提供一種神經移植體的製備方法，其 Ο 包括： [0009] S10，提供一培育層，所述培育層包括一疏水性基底、一 奈米碳管膜結構及一蛋白質層，所述奈米碳管膜結構設 置在所述矽膠基底，所述蛋白質層設置在該奈米碳管膜 結構表面； [0010] S20，在該蛋白質層表面種植複數神經細_ ;以及 Q [0011] S30，培育該複數神經細胞直到該複數神經細胞生長出複 數神經突起連接在所述複數神經細胞之間形成一神經網 路0 [0012] 在所述步驟S10中，所述奈米碳管膜結構由複數奈米碳管 組成。所述複數奈米碳管的延伸方向可基本平行於所述 奈米碳管膜結構的表面。優選地，所述複數奈米碳管之 間通過凡得瓦力（Van der Waals attractive force)連接，從而形成一自支撐結構。所謂“自支撐” 即該奈米碳管膜結構無需通過設置於一基體表面，亦能 099143857 表單編號A0101 第5頁/共40頁 0992075912-0 201224149 保持自身特定的形狀。由於該自支撐的奈米碳管膜結構 中大量的奈米碳管通過凡得瓦力相互吸引，從而使該奈 米碳管膜結構具有特定的形狀，形成一自支撐結構。所 述奈米碳管膜結構為自支撐結構時，該奈米碳管犋結構 可為由至少一奈米碳管膜形成的膜狀結構，當所述奈米 後管膜結構包括複數奈米碳管膜時，該複數奈米碳管犋 層疊設置，相鄰的奈米碳管膜之間通過凡得瓦力相結合 。由於所述奈米碳管膜基本由奈米碳管組成且奈米碳管 之間通過凡得瓦力連接，故所述奈米碳管膜結構具有彈 性佳、延展性良好及密度低等優點’便於裁剪和杈伸。 [0013] [0014] 請參閱圖2，所述奈米碳管膜可為一奈杀碳管絜化膜，該 奈米碳管絮化膜為將一奈米碳管原料囔化處理獲得的一 自支撐的奈米碳管膜。該奈米碳管絮化膜包括相互纏繞 且均勻分佈的奈米碳管。奈米碳管的長度大於10微米， 優選為200微米到900微米，從而使奈米碳管相互纏繞在 一起。所述奈米碳管之間通過凡得瓦力相互吸引、分佈 ’形成網路狀結構。由於該自支撐的奈米碳管絮化膜中 大量的奈米碳管通過凡得瓦力相互吸引並相互纏繞，從 而使該奈米碳管絮化膜具有特定的形狀，形成一自支撐 結構。所述奈米碳管絮化膜各向同性。所述奈米碳管絮 化膜中的奈米碳管為均勻分佈，無規則排列，形成大量 尺寸在1奈米到50 0奈米之間的間隙或微孔。所述間隙或 微孔能夠增加所述奈米碳管膜的比表面積及浸潤更多的 蛋白質。 所述奈米碳管膜可為一奈米碳管碾壓膜，該奈米碳管碾 099143857 表單編號A0101 第6頁/共40頁 0992075912-0 201224149 壓膜為通過碾壓一奈米碳管陣列獲得的一種具有自支撐 性的奈米碳管膜。該奈米碳管碾壓膜包括均勻分佈的奈 米碳管，奈米碳管沿同一方向或不同方向擇優取向排列 。所述奈米碳管碾壓膜中的奈米碳管相互部分交迭，並 通過凡得瓦力相互吸引，緊密結合，使得該奈米碳管膜 具有很好的柔韌性，可彎曲折迭成任意形狀而不破裂。 且由於奈米碳管碾壓膜中的奈米碳管之間通過凡得瓦力 相互吸引，緊密結合，使奈米碳管碾壓膜為一自支撐的 結構。所述奈米碳管碾壓膜中的奈米碳管與形成奈米碳 管陣列的生長基底的表面形成一夾角yS，其中，；5大於 等於0度且小於等於15度，該夾角/S與施加在奈米碳管陣 列上的壓力有關，壓力越大，該夾角越小，優選地，該 奈米碳管碾壓膜中的奈米碳管平行於該生長基底排列。 該奈米碳管碾壓膜為通過碾壓一奈米碳管陣列獲得，依 據碾壓的方式不同，該奈米碳管碾壓膜中的奈米碳管具 有不同的排列形式。具體地，奈米碳管可無序排列；請 參閱圖3，當沿不同方向碾壓時，奈米碳管沿不同方向擇 優取向排列；當沿同一方向碾壓時，奈米碳管沿一固定 方向擇優取向排列。該奈米碳管碾壓膜中奈米碳管的長 度大於50微米。 [0015] 該奈米碳管碾壓膜的面積與奈米碳管陣列的尺寸基本相 同。該奈米碳管碾壓膜厚度與奈米碳管陣列的高度以及 碾壓的壓力有關，可為0. 5奈米到1 0 0微米之間。可以理 解，奈米碳管陣列的高度越大而施加的壓力越小，則製 備的奈米碳管碾壓膜的厚度越大；反之，奈米碳管陣列 099143857 表單編號A0101 第7頁/共40頁 0992075912-0 201224149 的高度越小而施加的壓力越大，則製備的奈米碳管碾壓 膜的厚度越小。所述奈米碳管碾壓膜之中的相鄰的奈米 碳管之間具有一定間隙，從而在奈米碳管碾壓膜中形成 複數尺寸在1奈米到500奈米之間的間隙或微孔。所述間 隙或微孔能夠增加所述奈米碳管膜的比表面積及浸潤更 多的蛋白質。 [0016] 所述奈米碳管膜可為一奈米碳管拉膜，所述奈米碳管拉 膜係由若干奈米碳管組成的自支撐結構。請參閱圖4，所 述若干奈米碳_管為沿該奈米碳管拉膜的長度方向擇優取 向排列。所述擇優取向係指在奈米碳管拉膜中大多數奈 米碳管的整體延伸方向基本朝同一方向。而且，所述大 多數奈米碳管的整體延伸方向基本平行於奈米碳管拉膜 的表面。 [0017] 進一步地，所述奈米碳管拉膜中多數奈米碳管係通過凡 得瓦力首尾相連。具體地，所述奈米碳管拉膜中基本朝 同一方向延伸的大多數奈米碳管中每一奈米碳管與在延 伸方向上相鄰的奈米碳管通過凡得瓦力首尾相連。當然 ，所述奈米碳管拉膜中存在少數偏離該延伸方向的奈米 碳管，這些奈米碳管不會對奈米碳管拉膜中大多數奈米 碳管的整體取向排列構成明顯影響。所述自支撐為奈米 碳管拉膜不需要大面積的載體支撐，而只要相對兩邊提 供支撐力即能整體上懸空而保持自身膜狀狀態，即將該 奈米碳管拉膜置於（或固定於）間隔一定距離設置的兩 個支撐體上時，位於兩個支撐體之間的奈米碳管拉膜能 夠懸空保持自身膜狀狀態。所述自支撐主要通過奈米碳 099143857 表單編號A0101 第8頁/共40頁 0992075912-0 201224149 Ο 管拉膜中存在連續的通過凡得瓦力首尾相連延伸排列的 奈米碳管而實現。具體地，所述奈米碳管拉膜中基本朝 同一方向延伸的多數奈米碳管，並非絕對的直線狀，可 適當的彎曲；或者並非完全按照延伸方向上排列，可適 當的偏離延伸方向。故，不能排除奈米碳管拉膜的基本 朝同一方向延伸的多數奈米碳管中並列的奈米碳管之間 可能存在部分接觸。具體地，該奈米碳管拉膜包括複數 連續且定向排列的奈米碳管片段。該複數奈米碳管片段 通過凡得瓦力首尾相連。每一奈米碳管片段由複數相互 平行的奈米碳管組成。該奈米碳管片段具有任意的長度 、厚度、均勻性及形狀。該奈米碳管拉膜具有較好的透 光性，可見光透過率可達到75%以上。 [0018] ❹ 當該奈米碳管膜結構包括複數奈米碳管拉膜時，所述複 數奈米碳管拉膜層疊設置形成一層狀結構。該層狀結構 的厚度不限，相鄰的奈米碳管拉膜通過凡得瓦力結合。 優選地，所述層狀結構包括的奈米碳管膜的層數小於或 等於10層，從而使單位面積内的奈米碳管數量較少，使 該奈米碳管自身的拉曼光強保持在較小的範圍，從而減 小拉曼光譜中奈米碳管的拉曼峰強。該層狀結構中相鄰 的奈米碳管拉膜中的奈米碳管之間具有一交叉角度a， 且該α大於0度且小於等於90度。當相鄰的奈米碳管拉膜 中的奈米碳管之間具有一交叉角度α時，所述複數奈米 碳管拉膜中的奈米碳管相互交織形成一神經移植體，使 所述奈米碳管膜結構的機械性能增加。在本實施例中， 所述奈米碳管膜結構包括複數層奈米碳管拉膜層疊設置 099143857 表單編號Α0101 第9頁/共40頁 0992075912-0 201224149 相鄰的奈米碳官 [0019] [0020] 099143857 〜卡碳官之間的交又角 致等於90度’即’相鄰奈米碜管拉膜中的奈米碳政 伸方向大致平行。 、、反官的延 所述疏水性基底餘承載該所述奈米碳管膜結構及 質廣。所述疏水性基底具有疏水性及良好的㈣性。^ 述疏水性基底可切膠製成，或表面絲有㈣ I 疏水性基底的雜與厚度可拫據所述奈米碳管膜处 形狀與厚度設計性基底表面的二^ 形狀可大致與所述奈米碳管_構_觀形狀大2 當。可以理解，當所述奈米碳管膜結構的厚度較目 該奈米碳管膜結構具有較小機械強度及具有較大 故’該奈米碳管膜結構溶液受外力產 ^附在其他親水性物體上。將該奈米碳管膜結； 在所迷疏水性基底表面形成—生物基底時， ^機械強度㈣述奈米碳管麟構大，從太:底 官膜結構更難受外來作用而產生破損同時 防止該奈料管_構_在親水性物體上/移動及 _蛋白㈣設置麵述奈 使所迷太化〜目士 表面’用於 攻…管層具有親水性及生 ，培育層能夠為所述神經細胞_=而使得 管膜結構遠離所述疏水==設置在所述奈米後 白質層中的蛋白質（表面。優選地，所述蛋 白質可選自纖維狀蛋白質血^4蛋白f。所述蛋 在本實施例中’所辻尽 蛋白質及酶蛋白質等。 34蛋白質為哺乳動物的金清，如豬也 表卓編號A0101 ^ _ 第10頁/共40頁 201224149 清、牛血清或人血清。所述血清不僅能夠為所述神經細 胞的種植及生長提供—個合適的環境，還能夠在所述神 經細胞生㈣，為所述神經細胞提供 生長因數。 [0021] ❹ 〇 所述培育層的製備方料限，只要㈣使蛋白質層與所 述奈米碳管膜結構混合在_起即可。譬如，可通過將所 述奈米碳f膜結構浸泡在_蛋白質溶液中，使所述蛋白 質岭液浸潤所述奈米碳管膜結構，從而使得所述蛋白質 溶液中的蛋白質附著在所述奈米碳管膜結構遠離所述疏 水性基底的表面形成蛋自質層。亦可將所述蛋白質溶液 喷塗在所述奈米讀彻構祕所紐水性基底的表面 ’使所述蛋白質層設置在該表面。還可將蛋:白質溶液滴 在所述奈轉管膜結構遠離所述疏水,底的表面再 採用甩膜的方式使所述蛋白f層設置在該表面。所述蛋 白質溶液除所述蛋白質外，還可包括溶解所述蛋白質的 生物媒介（biological me(jia)，所述生物媒介的種類 不限，可根據蛋白質的種類的;^同而調製。通常，所述 蛋白質溶液中的蛋今質的濃度大:於等於50%小於等於1〇〇% 。在本實施例中，所述蛋白質溶液中的蛋白質濃度為 100%，即，所述蛋白質溶液為純蛋白質，無需溶劑溶解 [0022] 由於所述奈米碳管膜結構包括複數奈米碳管且複數奈米 碳管之間存在間隙形成複數微孔，當所述蛋白質溶液浸 潤所述奈米碳管膜結構時，所述蛋白質溶液可滲透入所 述奈米碳管膜結構内部浸潤所述複數奈米碳管的表面。 當然，在所述培育層中，並不係所有奈米碳管的表面均 099143857 表單編號A0101 第II頁/共40頁 0992075912Ό 201224149 可浸潤有蛋白質層，然，只需位於需培育神經細胞的奈 米碳管膜結構的表面的部分奈米碳管浸潤有蛋白質溶液 ，即可在所述奈米碳管膜結構表面形成蛋白質層，使所 述奈米碳管膜結構具有親水性及生物相容性，實現使培 育層具有作為神經細胞生長的載體的功能。這係因為， 由於奈米碳管具有疏水性，由奈米碳管組成的奈米碳管 膜結構並不能為神經細胞生長提供適合的親水性環境。 而當奈米碳管表面覆蓋有具有親水性及無毒性的蛋白質 層後，由覆蓋有蛋白質的奈米碳管組成的結構即能為細 胞生長提供適合的親水性環境。在本實施例中，所述培 育層的製備方法可進一步包括如下步驟： [0023] 在本實施例中，所述培育層的製備方法可進一步包括如 下步驟： [0024] S11，提供所述疏水性基底； [0025] S12，將所述奈米碳管膜結構設置在所述疏水性基底表面 9 [0026] S1 3，使所述奈米碳管膜結構浸潤有蛋白質溶液；以及 [0027] S1 4，對浸潤有蛋白質溶液的奈米碳管膜結構進行滅菌處 理形成所述培育層。 [0028] 在步驟S11中，所述疏水性基底由矽膠製成，或表面塗敷 有矽膠。矽膠為生物體常用的植入材料，對生物體無毒 性，且具有較好的柔韌性。故，由該矽膠基底製備形成 或者塗敷有矽膠的疏水性基底可直接植入人體。 099143857 表單編號A0101 第12頁/共40頁 0992075912-0 201224149 [0029]在步驟S12中，為使所述奈米碳管膜結構與所述矽膠基底 表面結合更緊密，可對所述奈米碳管膜結構進行有機溶 劑處理。具體地，可在設置在所述矽膠基底表面的奈米 碳管膜結構覆蓋或者滴上容易揮發的溶劑，如有機溶劑 ，再使所述溶劑揮發，從而可減小該奈米碳管獏結構的 比表面及增加該奈米碳管膜結構與所述矽膠基底的附著 力。 [0030] 〇 〇 [0031] 099143857 在步驟S13巾，使所述奈米碳管赌構浸财蛋自質溶液 的方式不限’只要使蛋白質溶液中的蛋白質附著在奈米 碳管膜結構表面形成一蛋白贊春即竒如，可通過將 所述奈米碳管膜結構浸泡在所述蛋白質溶液中，實現蛋 白質溶液的㈣。亦可通過在所杉^㈣結構喷塗 所述蛋Μ溶液，實現蛋白質溶液的浸潤《在本實施例 中’為實現蛋”溶液的㈣，選擇賴述奈米碳管膜 結構浸泡在純蛋白。所述奈米碳管麟構的浸泡時 間依奈米碳管膜結_具餘構及蛋白f溶液的具體組 分而定，只要能使所述奈米碳料結構中的大部分奈米 Γ浸潤有蛋”溶㈣可。通常，所述奈米碳管膜結 的浸泡時間在2小時以上。所述奈米碳管膜結構的浸泡 展境不限，只要不使所述蛋白f變質即可。通常，所 述浸泡過程可在常溫、常壓環境下進行。 =述奈結構浸泡在所述蛋白質溶液中時，所 ^白質溶液可浸_述奈米碳⑼結構中的部分奈米 碳^亦可浸潤所述奈米碳管膜結構中的全部奈米碳管 ，當所衫米碳管膜結構的厚度較薄時 ，'譬如 表單編號A0101 第13頁/共40頁 0992075912-0 201224149 所述奈米碳管膜結構的厚度小於等於1 〇微米時，所述蛋 白質溶液可浸潤所述奈米碳管膜結構中的全部奈米碳管 。當所述奈米碳管膜結構的厚度較厚時，譬如所述奈米 碳管膜結構的厚度大於等於10微米時，所述蛋白質溶液 可浸潤所述奈米碳管膜結構中遠離所述疏水性基底部分 的奈米碳管。需要指出的係，所述蛋白質溶液係否浸潤 所述奈米碳管膜結構中的全部奈米碳管除了與奈米碳管 膜結構的厚度有關外，還與浸潤時間與所述蛋白質溶液 的濃度有關。譬如，當浸潤時間較短時，即便所述奈米 碳管膜結構較薄，所述蛋白質溶液亦可能僅浸潤所述奈 米碳管膜結構中的部分奈米碳管。 [0032] 在步驟S14中，對浸潤有蛋白質溶液的奈米碳管膜結構進 行滅菌處理的方式不限，只要能夠殺死蛋白質溶液中的 大部分細菌即可。譬如可採用高溫滅菌或紫外光滅菌的 方式對所述蛋白質溶液進行滅菌。在本實施例中，採用 高溫殺菌的方式對該蛋白質溶液進行滅菌。當然，為使 所述蛋白質溶液中的蛋白質不至於被破壞，高溫滅菌時 的溫度不得超過220度。在本實施例中，所述高溫滅菌時 的溫度大致為120度。可以理解，當所述蛋白質溶液中本 身細菌較少，則該步驟S14則可省略。當對浸潤在奈米碳 管膜結構中的蛋白質溶液進行滅菌處理時，所述蛋白質 溶液中的溶劑或水分將減少。通常地，浸潤在所述奈米 碳管膜結構中的蛋白質溶液隨著溶劑或水分的減少而固 化，從而在所述奈米碳管膜結構的表面形成所述蛋白質 層。 099143857 表單編號Α0101 第14頁/共40頁 0992075912-0 201224149 [0_纟步驟S1G中’為增加該培育層對神經細胞的附著性及提 供更適合神經細胞的生長環境，在形成蛋白質層後，气 步驟sio還可進一步包括如下步驟：S15，在所述蛋白^ 層遠離所述疏水性基底的表面形成—聚賴氣酸（p〇ly D-lysine， PDL)層。 [0034] 在步驟s2〇中’所述神經細胞包括哺乳動物的神經細皰， 優選地，所述神經細胞為海馬神經元。在該培育層表面 種植複數神經細胞的方法不限，可採用在該培育層遠離 〇 所述疏水性基底的表面喷射或塗覆含有該神經細胞的溶 液’亦可採用將該:鸿育::層浸泡在所述含神經細胞的神經 細胞液中’只要使所述神經鰣胞液覆蓋所述培育層即可 。為使所述神經細胞液覆蓋所述培育層.，所述神經細胞 液可盛放在一培養皿中。所述培育層可懸空設置在所述 培養皿中。亦可設置在所述培養JBL的一底面上，只要能 使所述培養液覆蓋所述神經細胞即可。當所述培育層設 置在所述培養皿的底面時，所述疏水性基底與所述底面 〇 接觸’從而使得所述神經細胞僅分佈在蛋白質層表面。 [0035] 在步驟S30中’所述神經細胞的培育環境不限，只要能夠 生長出神經突起即可。通常，所述神經細胞在常溫、常 麼環境中即可生長。即，將所述神經細胞放置在室内環 境中，所述神經細胞即可生長，而培育層中的蛋白質如 牛jk清可提供生長因數’促進該神經細胞生長。當然， 亦可使該培a環境接近提供該神經細胞的生物體的體内 生長環境亦可。譬如’當所述神經細胞為取自老鼠的海 馬神經細胞時，可模幾所述老鼠體内的生長環境。 0992075912-0 099143857 表單編號如第15耳/并40頁 201224149 [0036] 所述神經細胞在培育時，能夠長出複數神經突起（N e u r -ite)。所述神經突起包括樹突（Dendrite)與軸突（ Axon )。當所述培育層表面僅有一個神經細胞時，所述 神經細胞的神經突起沿培育層的表面朝各個方向隨機生 長。然由於神經細胞本身會釋放出誘導神經突起定向生 長的因數，故，當所述培育層表面設置有複數神經細胞 時，該神經細胞的神經突起具有將沿向相鄰的神經細胞 生長的趨勢，從而使相鄰的神經細胞得以連接溝通。故 ，控制神經細胞在所述培育層的分佈，即可控制所述神 經突起的生長方向。譬如，如果所述神經細胞係隨機均 勻分佈在所述培育層的表面，所述神經細胞將各自生長 出神經突起與相鄰的細胞連接，當所述複數神經細胞的 全部或大多數神經細胞均生長出連接在相鄰的神經細胞 之間的神經突起時，所述複數神經細胞借由所述神經突 起形成所述神經網路，使該複數神經細胞之間能相互溝 通。相鄰的神經細胞的神經突起如果相遇，則會合為同 一個神經突起。再譬如，當所述神經細胞在該培育層表 面以線狀或者陣列的方式排列時，且沿縱向方向的神經 乡田胞相距較近，而沿橫向方向的神經細胞相距較遠，此 時，所述神經細胞所生長的神經細胞可基本沿所述縱向 方向延伸。為使所述神經細胞能夠在所述培育層表面以 線狀或者陣列的方式排列，可選擇使所述奈米碳管膜中 的奈米碳管基本沿同一方向延伸。通過培育，彼此相鄰 的神經細胞大多通過神經突起建立起連接，從而形成所 述神經網路。所述神經網路與所述培育層一起形成所述 神經移植體。 099143857 表單編號A0101 第16頁/共40頁 0992075912-0 201224149 [0037]所述移植體的製備方法通過在該疏水性基底表面設置奈 米碳管膜結構，再在該奈米碳管膜結構遠離所述疏水性 基底表面設置蛋白質層形成培育層，從而能在該培育層 中蛋白質層表面種植複數神經細胞，並使所述複數神經 細胞生長出複數神經突起連接起複數神經細胞。所述矽 膠基底具有疏水性，不能使所述神經細胞吸附其上，即 不能提供供所述神經細胞生長的環境，故，所述神經細 胞將僅在所述設置有蛋白質層表面生長。 〇 [〇〇38]所述奈米碳管膜結構為一宏觀的膜狀結構，其面積一般 都可達到15毫米xi5毫米以上，具體地，該奈求碳管膜結 構的長度可達300米以上，寬丨度可達〇.5米以上。所述疏 水性基底亦為一宏觀結構，莠形狀與面囌可根據所述奈 米碳管膜結構的形狀與面積進行調整。且該奈米碳管膜 結構與所述疏水性基底均具有彈性佳、延展性良好及不 含金屬等優點，可直接植八生物潑。故，由所述奈米碳 管膜結構及疏水性基底做主要載艟的神經移植體可根據 〇 爻損神經系統的受損部位的形狀、大小進行裁剪、拉伸 並植入受損部位。所述神經網路具有生物活性及信號傳 遞能力，從而使得包括所述神經網路的神經移植體亦具 有生物活性及仏號傳遞能力。當所述神經移植體植入生 物體中的受損部位時，由於所述神經植入體中的神經元 與所述受損部位兩端或邊緣的神經元的距離較短，故可 通過直接縫合所述神經植入體中的神經元與受損部位邊 緣的神經元的方式使所述受損部位的兩端建立起信號傳 遞能力，完成受損部位的神經修復，從而節省所述神經 099143857 表單編號A0101 第17頁/共40頁 0992075912-0 201224149 突起的生長時間，減少受損的神經系統的修復時間。可 以理解，即便係在所述神經植入體植入受損部位時，不 進行直接縫合，由於所述神經植入體中的神經元所述受 損部位邊緣的神經元的距離小於所述受損部位兩端的神 經元的距離，故，通過植入所述神經植入體，亦能減少 神經突起的生長時間，從而減少受損的神經系統的修復 時間。 [0039] 需要指出的係，通常情況下，所述奈米碳管膜結構中的 奈米碳管係指未經過化學或物理處理的奈米碳管，如未 經過表面親水性處理的奈米碳管，即，所述奈米碳管為 純奈米碳管。當然，奈米碳管膜結構中的奈米碳管如果 係經過改性的奈米碳管，只要係對神經細胞沒有毒性， 亦應在在本發明的保護範圍之内，只係，所述奈米碳管 的改性並不會對實現本發明有任何實質性貢獻，因為， 當所述蛋白質層覆蓋該奈米碳管後，所述神經細胞與所 述奈米碳管並不直接接觸，該奈米碳管的表面結構實際 上係可忽略的。 [0040] 本發明提供的神經移植體可包括由上述神經移植體的製 備方法在包括奈米碳管膜結構及蛋白質層的培育層表面 培養由複數神經細胞及神經突起形成的神經網路所得到 產品。 [0041] 請參閱圖5，所述神經移植體100包括一培育層10及分佈 在該培育層10表面的一神經網路20。 [0042] 所述培育層1 0包括一疏水性基底11、一奈米碳管膜結構 099143857 表單編號A0101 第18頁/共40頁 0992075912-0 201224149 [0043] Ο [0044] 〇 [0045] 099143857 12及一蛋白質層14。所述奈米碳管膜結構ι2設置在所述 疏水性基底11的一個或者相對的兩個表面。所述蛋白質 層14设置在所述奈米碳管膜結構丨2遠離所述疏水性基底 11的表面。在本實施例中，所述奈米碳管膜結構12僅設 置在所述疏水性基底11的一個表面。 所述疏水性基底11可由矽膠製成，或表面塗敷有矽膠。 矽膠為生物體常用的植入材料，對生物體無毒性，且具 有較好的柔韌性。故，由該矽膠製備形成或者塗敷有矽 膠的疏水性基底11可直接植入人體。 所述奈来碳管膜結構12包括複數奈米碳管基本平行於所 述奈米碳管膜結構的表面，且相鄰的奈'米碳管之間通過 凡得瓦力相互連接形成—自支撐結構 所述奈米碳管膜 結構12包括至少一奈米碳管膜，該奈米碳管膜可為如圖2 中的奈米碳管絮化膜、圖3中的奈米碳營碾壓膜及圖4中 的奈米碳管拉膜《在本實施例中，所述奈米碳管膜結構 12包括複數層疊設置的拉膜，相辦的拉膜通過凡得瓦力 相互結合。在相鄰的拉膜中，奈米碳管的的延伸方向可 具有一個交又角度，優選地，所述交又角度為9〇度。所 述奈米碳管膜結構12的厚度可根據具體需求而設置。通 常，所述奈米碳管膜結構12厚度大於〇. 3微米小於6〇微米 。在本實施例中，所述奈米碳管膜結構12的厚度大致為 0. 6微米。 所述蛋白質層丨4為由可溶性蛋白質組^所謂可溶性蛋 白質即該蛋白質具有較好的親水性。所述蛋白質層14的 厚度不限，只要能夠提供—偷如， ^個親水性環境即可。通當， 表單編號Α0101 第19頁/共4〇頁 中 0992075912-0 201224149 所述蛋白質層14的厚度极3微㈣2微米。在本實施例 中，所述蛋白質層的厚度大致為〇· 5微米。在宏觀上，所 述蛋白質層U可選難設置在所述奈米碳㈣結構㈣ 離所述疏水性基底11的表面。在微觀上，所述蛋白質層 14中的蛋白質谷易參透到所述奈米碳管膜結構12的内部 ^包覆所述奈米碳管膜結構12中的部分或者全部奈米 石反S ’此時’所述蛋白質層14與該奈米碳管膜結構12之 間並沒有明顯的分介面。通常，當所述奈米碳管膜結構 12的厚度較薄時，譬如，所迷奈米碳管膜結構上2的厚度 小於等於3微米時，所述蛋白質層Π中的蛋白質容易滲透 到所述奈米碳管膜結構12的崎，並基本包覆所述夺米 碳管膜結構12中的所有的奈米碳管。而當所述奈米碳管 膜結構12的厚度較厚時，譬如，所述奈米碳管膜結構12 的厚度大於等於3微米時，所述蛋白質㈣中的蛋白質雖 然亦可滲透到所述奈米碳管膜結構12内部1通 覆所述奈米碳管膜結構_ 網路2 0的奈米碳 管。在本實_中，所述蛋白質層14中的蛋白質基本包 覆所述奈米碳管膜結構12中的所有的奈米碳管。 [0046] 所述神經網路20設置在所述蛋白質層u遠離所述奈米碳 管膜結構12的—個表面。當所述神經移植體m僅包括_ 個奈米碳管赌構12且該奈米碳管麟構12_疏水性 基底u的表面設置有一個蛋白質層14時，所述神經移植 體100僅包括-個神經網路2G設置在所述蛋白質層Μ表面 。當所述神經移植㈣〇包括兩個奈米碳切結構12分別 设置在該疏水性基底11的兩個表面m+碳" 099143857 表單編號A0101 第20頁/共4〇頁 0992075912-0 201224149 [0047] Ο [0048][0003] 099143857 In the system, neurons receive treatment to receive stimulation, conduct and output neurotransmitters (neuron _tr.ansmi tter) for tissue or organ communication, and glial cells perform neurophysical physics Sexual support, nutrition provision, and adjustment of the speed of the communication base. Each neuron depends on the type of cell body and neurite. The neurites extend from the cell body and grow toward other neurons or other cells (eg, muscle cells), where the neurites Divided into axons Ux〇n) and dendrites (dendrite). In general, the stimulus is received by the tree and transmits the impulse to the cell clock. The impulse is transmitted through the axon to the end of the axon, and the micro-materials come to other cells. Since the nervous system plays a coordinating role between tissues and organs in the living body, its importance is not lost. Previously, neurological deficits due to impaired nervous system were caused by temporary riding (4). Among them, the damaged nervous system is repaired by implanting the H hybrid, and the neurosurgery is used to repair all kinds of emotions (4) and (4) (4) to damage the money. Previous nerve grafts were usually the nerve tubes at both ends of the nerve, and the gods of the (four) system were made of biodegradable materials. Form No. A0101 Page 3 of 40 0992075912-0 201224149 [0004] [ 0005] [0006] [0007] 099143857 structure. The nervous system damaged part 1 (four) "_ inner tube (four) Μ (four)" through the other end of the damaged part of the system. Usually, it needs to be repaired by implanting the god (four) The length of the damaged part is longer, and the repair time required for repairing the damaged nervous system by the neurological officer of the Zhouzhou is longer. [Invention content] Reduction of __ systemic repair time Shen 1 The preparation method of the transplant body is actually Qin Qin. - The kind of nerve graft, including "hydrophobic base, "Nie obstructive membrane structure, a protein layer and - neural network The carbon nanotube film structure is disposed on the surface of the hydrophobic substrate, and the protein layer is disposed on a surface of the carbon nanotube bottom. The neural network is disposed at The protein layer is remote from the carbon nanotube The surface of the structure describes the neural network including a plurality of nerve cells and a plurality of neurites. The neurites between the plurality of nerve cells are connected to each other to form a neural network. Compared with the technique of the Lin technology Forming the neural network on the surface of the cultivating layer. The carbon nanotube membrane structure and the hydrophobic substrate in the cultivating layer have advantages such as good elasticity and good ductility, and the nerve graft can be subjected to The shape and size of the damaged part of the damaged nervous system are cut, stretched, and damaged. The neural network has biological activity and signal transmission capability, so that the nerve graft including the neural network also has Biological activity and signal transmission ability. When the surface is numbered Α0101 Page 4/40 pages 0992075912-0 201224149 When the nerve graft is implanted into the damaged part of the organism, due to the neurons in the nerve implant The distance from the neurons at the ends or edges of the damaged portion is very close, so that the neurons in the nerve implant and the neurons at the edge of the damaged portion can be directly sutured. The method establishes a signal transmission capability at both ends of the damaged portion, completes nerve repair of the damaged portion, thereby saving growth time of the neurite, and reducing repair time of the damaged nervous system. [Embodiment] _ [ 0008] Referring to FIG. 1 , the present invention provides a method for preparing a nerve graft, which includes: [0009] S10, providing a cultivating layer, the cultivating layer comprising a hydrophobic substrate, a carbon nanotube film structure, and a protein layer, the carbon nanotube membrane structure is disposed on the silicone substrate, the protein layer is disposed on the surface of the carbon nanotube membrane structure; [0010] S20, implanting a plurality of neuronal fines on the surface of the protein layer; And Q [0011] S30, cultivating the plurality of nerve cells until the plurality of nerve cells grow a plurality of neurites to form a neural network between the plurality of nerve cells. [0012] In the step S10, the nai The carbon nanotube membrane structure is composed of a plurality of carbon nanotubes. The plurality of carbon nanotubes may extend substantially parallel to the surface of the carbon nanotube membrane structure. Preferably, the plurality of carbon nanotubes are connected by a Van der Waals attractive force to form a self-supporting structure. The so-called "self-supporting" means that the carbon nanotube film structure does not need to be disposed on a surface of a substrate, and can also be maintained in its own specific shape by 099143857 Form No. A0101 Page 5 of 40 0992075912-0 201224149. Since a large number of carbon nanotubes in the self-supporting carbon nanotube membrane structure are attracted to each other by van der Waals force, the carbon nanotube membrane structure has a specific shape to form a self-supporting structure. When the carbon nanotube membrane structure is a self-supporting structure, the carbon nanotube structure may be a film-like structure formed by at least one carbon nanotube film, and when the nano-back membrane structure comprises a plurality of nanometers In the case of the carbon nanotube film, the plurality of carbon nanotubes are stacked, and the adjacent carbon nanotube films are combined by van der Waals force. Since the carbon nanotube film is basically composed of a carbon nanotube and the carbon nanotubes are connected by van der Waals, the carbon nanotube film structure has the advantages of good elasticity, good ductility and low density. Easy to cut and stretch. [0014] Referring to FIG. 2, the carbon nanotube film may be a carbon nanotube film, and the carbon nanotube film is obtained by deuterating a carbon nanotube material. A self-supporting carbon nanotube film. The carbon nanotube flocculation membrane comprises carbon nanotubes which are intertwined and uniformly distributed. The carbon nanotubes have a length greater than 10 microns, preferably from 200 microns to 900 microns, such that the carbon nanotubes are intertwined with each other. The carbon nanotubes are mutually attracted and distributed by van der Waals to form a network structure. Since the large number of carbon nanotubes in the self-supporting carbon nanotube flocculation membrane are attracted to each other and entangled by van der Waals force, the carbon nanotube flocculation membrane has a specific shape to form a self-supporting structure. . The carbon nanotube flocculation membrane is isotropic. The carbon nanotubes in the carbon nanotube film are uniformly distributed and randomly arranged to form a large number of gaps or micropores having a size ranging from 1 nm to 50 nm. The gap or micropores can increase the specific surface area of the carbon nanotube membrane and infiltrate more protein. The carbon nanotube film can be a carbon nanotube rolled film, the carbon nanotube mill 099143857 Form No. A0101 Page 6 / Total 40 page 0992075912-0 201224149 The film is passed through a carbon nanotube A self-supporting carbon nanotube film obtained from the array. The carbon nanotube rolled film comprises uniformly distributed carbon nanotubes, and the carbon nanotubes are arranged in the same direction or in different directions. The carbon nanotubes in the carbon nanotube rolled film partially overlap each other and are attracted to each other by van der Waals force, and the carbon nanotube film has good flexibility and can be flexibly folded. In any shape without breaking. Moreover, since the carbon nanotubes in the carbon nanotube rolled film are attracted to each other by the van der Waals force, the carbon nanotube film is a self-supporting structure. The carbon nanotubes in the carbon nanotube rolled film form an angle yS with the surface of the growth substrate forming the carbon nanotube array, wherein; 5 is greater than or equal to 0 degrees and less than or equal to 15 degrees, the angle / S In relation to the pressure exerted on the carbon nanotube array, the larger the pressure, the smaller the angle, and preferably, the carbon nanotubes in the carbon nanotube rolled film are aligned parallel to the growth substrate. The carbon nanotube rolled film is obtained by rolling a carbon nanotube array, and the carbon nanotubes in the carbon nanotube rolled film have different arrangements depending on the manner of rolling. Specifically, the carbon nanotubes can be arranged in disorder; referring to FIG. 3, when rolling in different directions, the carbon nanotubes are arranged in different orientations; when crushed in the same direction, the carbon nanotubes are along a The orientation is preferred and the orientation is preferred. The length of the carbon nanotubes in the carbon nanotube rolled film is greater than 50 microns. [0015] The area of the carbon nanotube rolled film is substantially the same as the size of the carbon nanotube array. The thickness of the carbon nanotube film is in relation to the height of the carbon nanotube array and the pressure of the rolling, and may be between 0.5 nm and 100 μm. It can be understood that the larger the height of the carbon nanotube array and the lower the applied pressure, the greater the thickness of the prepared carbon nanotube rolled film; otherwise, the carbon nanotube array 099143857 Form No. A0101 Page 7 / Total The smaller the height of the 40 pages 0992075912-0 201224149, the greater the pressure applied, and the smaller the thickness of the prepared carbon nanotube rolled film. There is a gap between adjacent carbon nanotubes in the carbon nanotube film, thereby forming a gap between 1 nm and 500 nm in the carbon nanotube film. Or micropores. The gap or micropores can increase the specific surface area of the carbon nanotube membrane and infiltrate more protein. [0016] The carbon nanotube film may be a carbon nanotube film, and the carbon nanotube film is a self-supporting structure composed of a plurality of carbon nanotubes. Referring to Figure 4, the plurality of nanocarbon tubes are arranged in a preferred orientation along the length of the carbon nanotube film. The preferred orientation means that the majority of the carbon nanotubes in the carbon nanotube film are oriented in substantially the same direction. Moreover, the overall direction of extension of the majority of the carbon nanotubes is substantially parallel to the surface of the carbon nanotube film. [0017] Further, most of the carbon nanotubes in the carbon nanotube film are connected end to end by van der Waals force. Specifically, each of the carbon nanotubes of the majority of the carbon nanotubes extending in the same direction in the carbon nanotube film is connected end to end with the carbon nanotubes adjacent in the extending direction by van der Waals force . Of course, there are a few carbon nanotubes in the carbon nanotube film that deviate from the extending direction. These carbon nanotubes do not constitute an obvious alignment of the majority of the carbon nanotubes in the carbon nanotube film. influences. The self-supporting carbon nanotube film does not require a large-area carrier support, and as long as the support force is provided on both sides, it can be suspended in the whole to maintain its own film state, that is, the carbon nanotube film is placed (or When fixed on two supports arranged at a certain distance, the carbon nanotube film located between the two supports can be suspended to maintain its own film state. The self-supporting mainly through the nano carbon 099143857 Form No. A0101 Page 8 / Total 40 pages 0992075912-0 201224149 Ο There is a continuous flow of carbon nanotubes in the tube film extending through the end of the van der Waals. Specifically, the plurality of carbon nanotubes extending substantially in the same direction in the carbon nanotube film are not absolutely linear and may be appropriately bent; or are not completely aligned in the extending direction, and may be appropriately deviated from the extending direction. . Therefore, it is not possible to exclude partial contact between the carbon nanotubes juxtaposed in the majority of the carbon nanotubes extending substantially in the same direction. Specifically, the carbon nanotube film comprises a plurality of continuous and aligned carbon nanotube segments. The plurality of carbon nanotube segments are connected end to end by van der Waals force. Each carbon nanotube segment consists of a plurality of carbon nanotubes that are parallel to each other. The carbon nanotube segments have any length, thickness, uniformity, and shape. The carbon nanotube film has good light transmittance and the visible light transmittance can reach more than 75%. [0018] When the carbon nanotube film structure comprises a plurality of carbon nanotube film, the plurality of carbon nanotube films are laminated to form a layered structure. The thickness of the layered structure is not limited, and the adjacent carbon nanotube film is bonded by van der Waals force. Preferably, the layered structure comprises a carbon nanotube film having a layer number of less than or equal to 10 layers, so that the number of carbon nanotubes per unit area is small, and the Raman light intensity of the carbon nanotube itself is made strong. It is kept in a small range, thereby reducing the Raman peak intensity of the carbon nanotubes in the Raman spectrum. The carbon nanotubes in the adjacent carbon nanotube film in the layered structure have an angle of intersection a, and the α is greater than 0 degrees and less than or equal to 90 degrees. When the carbon nanotubes in the adjacent carbon nanotube film have an intersection angle α, the carbon nanotubes in the composite carbon nanotube film are intertwined to form a nerve graft. The mechanical properties of the carbon nanotube membrane structure are increased. In this embodiment, the carbon nanotube film structure comprises a plurality of layers of carbon nanotubes laminated film set 099143857 Form No. 1010101 Page 9 / Total 40 pages 0992075912-0 201224149 Adjacent nano carbon official [0019] [0020] 099143857 ~ card carbon official between the angle of the angle is equal to 90 degrees 'that is, the direction of the nano carbon in the adjacent nano tube is roughly parallel. The anti-official extension of the hydrophobic substrate carries the structure and quality of the carbon nanotube film. The hydrophobic substrate has hydrophobicity and good (tetra) properties. ^ The hydrophobic substrate can be made by cutting rubber, or the surface silk has (4) I. The hydrophobicity and thickness of the hydrophobic substrate can be approximated according to the shape and thickness of the carbon nanotube film. The description of the carbon nanotubes _ structure _ view shape 2 when. It can be understood that when the thickness of the carbon nanotube film structure is smaller than that of the carbon nanotube film structure, and the structure has a larger mechanical strength, the carbon nanotube film structure solution is attached to other hydrophilic substances by external force. On a sexual object. The carbon nanotube film is formed; when the biological substrate is formed on the surface of the hydrophobic substrate, the mechanical strength (4) indicates that the carbon nanotube structure is large, and the structure of the nano-membrane is more difficult to be externally damaged. Preventing the tube _ _ _ on the hydrophilic object / moving and _ protein (four) setting the face to make the fascination ~ visceral surface 'for the attack... the tube layer is hydrophilic and raw, the cultivating layer can be Said nerve cell _= such that the tubular membrane structure is away from the hydrophobicity == protein disposed in the post-nano white matter layer (surface. Preferably, the protein may be selected from the group consisting of fibrous protein blood protein 4 In the present example, the egg is depleted of protein, enzyme protein, etc. 34 protein is a mammalian gold clear, such as pig also has the number A0101 ^ _ 10th page / a total of 40 pages 201224149 clear, bovine serum or human serum The serum not only provides a suitable environment for the growth and growth of the nerve cells, but also provides a growth factor for the nerve cells (4). [0021] 培育 〇 the culture layer Preparation limit, as long as (4) mixing the protein layer with the carbon nanotube membrane structure, for example, by soaking the nanocarbon f membrane structure in a protein solution, infiltrating the protein lysate into the naphthalene solution a carbon nanotube film structure such that proteins in the protein solution adhere to the surface of the carbon nanotube film structure away from the surface of the hydrophobic substrate to form an egg self-layer. The protein solution may also be sprayed on the substrate. Said nanometer reading the surface of the aqueous substrate to make the protein layer on the surface. The egg: white matter solution may also be dropped on the surface of the navel membrane away from the hydrophobic, bottom surface The membrane is disposed in such a manner that the protein f layer is disposed on the surface. The protein solution may include, in addition to the protein, a biological medium (biological me) in which the protein is dissolved, and the type of the biological medium is not limited. It can be prepared according to the kind of the protein; generally, the concentration of the egg mass in the protein solution is large: equal to 50% or less and 1% or less. In the present embodiment, the protein solution is Protein rich The degree is 100%, that is, the protein solution is pure protein, and no solvent is dissolved. [0022] Since the carbon nanotube membrane structure includes a plurality of carbon nanotubes and a gap exists between the plurality of carbon nanotubes to form a plurality of micropores When the protein solution infiltrates the carbon nanotube membrane structure, the protein solution may penetrate into the surface of the carbon nanotube membrane structure to infiltrate the surface of the plurality of carbon nanotubes. Of course, in the cultivation In the layer, the surface of all carbon nanotubes is not 099143857. Form No. A0101 Page II / Total 40 pages 0992075912Ό 201224149 It can be infiltrated with protein layer. However, it only needs to be located in the structure of the carbon nanotube membrane that needs to cultivate nerve cells. A part of the surface of the carbon nanotube is infiltrated with a protein solution to form a protein layer on the surface of the carbon nanotube membrane structure, so that the carbon nanotube membrane structure has hydrophilicity and biocompatibility, and the cultivation layer is realized. It has a function as a carrier for the growth of nerve cells. This is because, because of the hydrophobicity of the carbon nanotubes, the structure of the carbon nanotube membrane composed of carbon nanotubes does not provide a suitable hydrophilic environment for nerve cell growth. When the surface of the carbon nanotube is covered with a hydrophilic and non-toxic protein layer, the structure consisting of a carbon nanotube covered with protein can provide a suitable hydrophilic environment for cell growth. In this embodiment, the method for preparing the cultivating layer may further include the following steps: [0023] In the embodiment, the method for preparing the cultivating layer may further include the following steps: [0024] S11, providing the hydrophobic layer a substrate; [0025] S12, the carbon nanotube film structure is disposed on the surface of the hydrophobic substrate 9 [0026] S1 3, so that the carbon nanotube film structure is infiltrated with a protein solution; and [0027] S1 4, sterilizing the carbon nanotube membrane structure infiltrated with the protein solution to form the incubation layer. [0028] In step S11, the hydrophobic substrate is made of silicone or the surface is coated with silicone. Tannin is a commonly used implant material for living organisms, is non-toxic to organisms, and has good flexibility. Therefore, a hydrophobic substrate formed or coated with tannin by the silicone substrate can be directly implanted into the human body. 099143857 Form No. A0101 Page 12 / Total 40 Page 0992075912-0 201224149 [0029] In step S12, in order to make the carbon nanotube membrane structure more tightly bonded to the surface of the silicone substrate, the nanocarbon may be The membrane structure is treated with an organic solvent. Specifically, the carbon nanotube film structure disposed on the surface of the silicone substrate may be covered or dropped with a solvent which is easily volatilized, such as an organic solvent, and then the solvent is volatilized, thereby reducing the structure of the carbon nanotube structure. The specific surface area and the adhesion of the carbon nanotube film structure to the silicone substrate. [0030] 99[0031] 099143857 In step S13, the manner in which the carbon nanotubes are immersed in the self-quality solution of the egg is not limited as long as the protein in the protein solution is attached to the surface of the carbon nanotube membrane structure. Forming a protein Zanchun, for example, can be achieved by immersing the carbon nanotube membrane structure in the protein solution to achieve a protein solution (4). The infiltration of the protein solution can also be achieved by spraying the egg tart solution on the structure of the cedar ^ (4) "in the present embodiment, the solution of the egg is realized" (4), and the structure of the lyon nano carbon tube membrane is selected to be immersed in the pure protein. The soaking time of the carbon nanotube structure is determined by the specific components of the composition and the protein f solution, as long as the majority of the nano carbon material structure can be used. ΓInfiltrated with egg" dissolve (four). Typically, the carbon nanotube film has a soaking time of more than 2 hours. The soaking exhibition of the carbon nanotube film structure is not limited as long as the protein f is not deteriorated. Usually, the soaking process can be carried out under normal temperature and normal pressure conditions. When the structure is immersed in the protein solution, the white matter solution can be immersed, and a part of the nanocarbon in the nano carbon (9) structure can also infiltrate all the nano carbon in the carbon nanotube film structure. Tube, when the thickness of the carbon nanotube film structure is thin, '譬如 Form No. A0101 Page 13 / Total 40 Page 0992075912-0 201224149 When the thickness of the carbon nanotube membrane structure is less than or equal to 1 〇 micron, The protein solution can infiltrate all of the carbon nanotubes in the carbon nanotube membrane structure. When the thickness of the carbon nanotube film structure is thick, such as when the thickness of the carbon nanotube film structure is greater than or equal to 10 micrometers, the protein solution may infiltrate the carbon nanotube film structure away from the A carbon nanotube of a hydrophobic base portion. It should be noted that the protein solution is infiltrated by all the carbon nanotubes in the carbon nanotube membrane structure, in addition to the thickness of the carbon nanotube membrane structure, and the infiltration time and the protein solution. Concentration related. For example, when the infiltration time is short, even if the structure of the carbon nanotube film is thin, the protein solution may only infiltrate a part of the carbon nanotubes in the carbon nanotube film structure. [0032] In step S14, the manner in which the carbon nanotube membrane structure infiltrated with the protein solution is sterilized is not limited as long as most of the bacteria in the protein solution can be killed. For example, the protein solution can be sterilized by means of high temperature sterilization or ultraviolet light sterilization. In this embodiment, the protein solution is sterilized by high temperature sterilization. Of course, in order to prevent the protein in the protein solution from being destroyed, the temperature at the time of high temperature sterilization must not exceed 220 degrees. In the present embodiment, the temperature at the time of high temperature sterilization is approximately 120 degrees. It will be understood that this step S14 may be omitted when there are fewer bacteria in the protein solution. When the protein solution infiltrated in the carbon nanotube membrane structure is sterilized, the solvent or moisture in the protein solution will be reduced. Generally, the protein solution infiltrated in the structure of the carbon nanotube film is solidified as the solvent or moisture is reduced to form the protein layer on the surface of the carbon nanotube film structure. 099143857 Form No. Α0101 Page 14 of 40 0992075912-0 201224149 [0_纟Step S1G' in order to increase the adhesion of the layer to nerve cells and provide a more suitable growth environment for nerve cells, after forming a protein layer, The gas step sio may further comprise the step of: S15, forming a p〇ly D-lysine (PDL) layer on the surface of the protein layer away from the hydrophobic substrate. [0034] In step s2, the neural cells comprise neuropilin of a mammal, preferably the neural cells are hippocampal neurons. The method of implanting a plurality of nerve cells on the surface of the cultivating layer is not limited, and a solution containing the nerve cells may be sprayed or coated on the surface of the cultivating layer away from the hydrophobic substrate. The layer is immersed in the nerve cell-containing nerve cell fluid 'as long as the neural sputum cytosol covers the cultivating layer. In order for the nerve cell fluid to cover the culture layer, the nerve cell fluid may be contained in a culture dish. The incubation layer can be suspended in the culture dish. It may be provided on a bottom surface of the culture JBL as long as the culture solution can cover the nerve cells. When the incubation layer is disposed on the bottom surface of the culture dish, the hydrophobic substrate is in contact with the bottom surface ’ such that the nerve cells are only distributed on the surface of the protein layer. [0035] In step S30, the incubation environment of the nerve cells is not limited as long as the neurites can be grown. Usually, the nerve cells can grow in a normal temperature environment. That is, the nerve cells are placed in an indoor environment, and the nerve cells can be grown, and proteins in the culture layer such as bovine jk clear can provide a growth factor to promote the growth of the nerve cells. Of course, the environment of the culture can also be brought close to the growth environment of the organism providing the nerve cells. For example, when the nerve cell is a hippocampal nerve cell taken from a mouse, the growth environment in the mouse can be molded. 0992075912-0 099143857 The form number is, for example, the 15th ear/and the 40th page 201224149 [0036] The nerve cells are capable of growing a plurality of neurites (N e u r -ite) when incubated. The neurites include dendrites and axons (Axon). When there is only one nerve cell on the surface of the growth layer, the nerve cells of the nerve cell grow randomly along the surface of the growth layer in various directions. However, since the nerve cells themselves release a factor that induces directional growth of the neurites, when the plurality of nerve cells are disposed on the surface of the growth layer, the nerve cells of the nerve cells have a tendency to grow along adjacent nerve cells. Thereby the adjacent nerve cells can be connected and communicated. Therefore, by controlling the distribution of nerve cells in the growth layer, the growth direction of the nerve protrusions can be controlled. For example, if the neural cell line is randomly and evenly distributed on the surface of the incubation layer, the nerve cells will each grow a neurite out to connect with adjacent cells, when all or most of the nerve cells of the plurality of nerve cells are When a neurite is connected between adjacent nerve cells, the plurality of nerve cells form the neural network by the neurites, so that the plurality of nerve cells can communicate with each other. If the neurites of adjacent nerve cells meet, they will merge into the same neurite. For example, when the nerve cells are arranged in a line or array on the surface of the growth layer, and the nerve cells in the longitudinal direction are relatively close, and the nerve cells in the lateral direction are far apart, at this time, The nerve cells grown by the nerve cells may extend substantially in the longitudinal direction. In order to enable the nerve cells to be arranged in a line or array on the surface of the growth layer, it is optional to extend the carbon nanotubes in the carbon nanotube film substantially in the same direction. By cultivating, nerve cells adjacent to each other are mostly connected by neurites to form the neural network. The neural network forms the nerve graft together with the incubation layer. 099143857 Form No. A0101 Page 16 / Total 40 Page 0992075912-0 201224149 [0037] The method for preparing a transplant body is provided by disposing a carbon nanotube membrane structure on the surface of the hydrophobic substrate, and then moving away from the structure of the carbon nanotube membrane The surface of the hydrophobic substrate is provided with a protein layer to form a cultivating layer, so that a plurality of nerve cells can be implanted on the surface of the protein layer in the cultivating layer, and the plurality of nerve cells are grown to form a plurality of neurites connected to the plurality of nerve cells. The silicone substrate is hydrophobic and does not allow the nerve cells to adsorb thereon, i.e., does not provide an environment for the growth of the nerve cells, so that the nerve cells will only grow on the surface where the protein layer is disposed.奈[〇〇38] The structure of the carbon nanotube membrane is a macroscopic membrane-like structure, and its area can generally reach 15 mm xi5 mm or more. Specifically, the length of the carbon nanotube membrane structure can reach 300 m. Above, the width can reach more than 米5 meters. The hydrophobic substrate is also a macroscopic structure, and the shape and surface of the crucible can be adjusted according to the shape and area of the carbon nanotube film structure. Moreover, the structure of the carbon nanotube film and the hydrophobic substrate have the advantages of good elasticity, good ductility and no metal, and can directly implant eight organisms. Therefore, the nerve graft mainly composed of the carbon nanotube membrane structure and the hydrophobic substrate can be cut, stretched, and implanted into the damaged portion according to the shape and size of the damaged portion of the sacral nerve system. The neural network has biological activity and signal transmission capabilities such that the neural graft including the neural network also has biological activity and nickname transmission ability. When the nerve graft is implanted into a damaged part of the living body, since the distance between the neuron in the nerve implant and the neurons at both ends or edges of the damaged part is short, it can be directly The manner in which the neurons in the nerve implant and the neurons at the edge of the damaged portion are sutured establishes a signal transmission capability at both ends of the damaged portion, and nerve repair of the damaged portion is completed, thereby saving the nerve 099143857 Form No. A0101 Page 17 of 40 0992075912-0 201224149 The growth time of the protrusions reduces the repair time of the damaged nervous system. It can be understood that even when the nerve implant is implanted into the damaged site, no direct suturing is performed, because the distance of the neurons at the edge of the damaged portion of the neuron in the nerve implant is smaller than the received The distance between the neurons at both ends of the lesion is reduced. Therefore, by implanting the nerve implant, the growth time of the neurites can also be reduced, thereby reducing the repair time of the damaged nervous system. [0039] It should be noted that, in general, the carbon nanotubes in the carbon nanotube membrane structure refer to carbon nanotubes that have not been subjected to chemical or physical treatment, such as nanoparticles that have not been subjected to surface hydrophilic treatment. The carbon tube, that is, the carbon nanotube is a pure carbon nanotube. Of course, if the carbon nanotubes in the carbon nanotube membrane structure are modified carbon nanotubes, as long as they are not toxic to nerve cells, it should be within the scope of the present invention. The modification of the carbon nanotubes does not have any substantial contribution to the realization of the present invention because, after the protein layer covers the carbon nanotubes, the nerve cells are not in direct contact with the carbon nanotubes. The surface structure of the carbon nanotube is actually negligible. [0040] The nerve graft provided by the present invention may include a neural network formed by a plurality of nerve cells and neurites on the surface of the culture layer including the carbon nanotube membrane structure and the protein layer by the preparation method of the above nerve graft. product. Referring to FIG. 5, the nerve graft 100 includes a seed layer 10 and a neural network 20 distributed on the surface of the seed layer 10. [0042] The incubation layer 10 includes a hydrophobic substrate 11, a carbon nanotube film structure 099143857 Form No. A0101 Page 18 / Total 40 Page 0992075912-0 201224149 [0043] Ο [0044] 〇 [0045] 099143857 12 and a protein layer 14. The carbon nanotube film structure ι2 is disposed on one or opposite surfaces of the hydrophobic substrate 11. The protein layer 14 is disposed on a surface of the carbon nanotube film structure 丨 2 away from the hydrophobic substrate 11. In the present embodiment, the carbon nanotube film structure 12 is provided only on one surface of the hydrophobic substrate 11. The hydrophobic substrate 11 may be made of silicone or coated with silicone. Tannin is a commonly used implant material for living organisms, is non-toxic to organisms, and has good flexibility. Therefore, the hydrophobic substrate 11 formed or coated with the silicone rubber can be directly implanted into the human body. The carbon nanotube membrane structure 12 includes a plurality of carbon nanotubes substantially parallel to the surface of the carbon nanotube membrane structure, and adjacent naphtha carbon nanotubes are formed by mutual connection of van der Waals forces. Support structure The carbon nanotube membrane structure 12 comprises at least one carbon nanotube membrane, which may be a carbon nanotube flocculation membrane as shown in FIG. 2, and a nano carbon cylinder in FIG. The film and the carbon nanotube film of FIG. 4 In the present embodiment, the carbon nanotube film structure 12 includes a plurality of laminated films, and the corresponding film is bonded to each other by van der Waals force. In the adjacent drawn film, the extending direction of the carbon nanotubes may have an intersection angle, and preferably, the intersection angle is 9 degrees. The thickness of the carbon nanotube film structure 12 can be set according to specific needs. Typically, the carbon nanotube film structure 12 has a thickness greater than 0.3 microns and less than 6 microns. 6微米。 The thickness of the carbon nanotube film structure 12 is substantially 0.6 microns. The protein layer 丨4 is composed of a soluble protein group, so-called soluble protein, that is, the protein has good hydrophilicity. The thickness of the protein layer 14 is not limited as long as it can provide a stealing environment. Properly, Form No. Α0101 Page 19 of 4 Page 0992075912-0 201224149 The thickness of the protein layer 14 is extremely 3 micro (four) 2 microns. In this embodiment, the thickness of the protein layer is approximately 〇·5 μm. Macroscopically, the protein layer U may be difficult to arrange on the surface of the nanocarbon (tetra) structure (4) from the hydrophobic substrate 11. Microscopically, the protein valleys in the protein layer 14 are easily penetrated into the interior of the carbon nanotube membrane structure 12 to coat part or all of the nano-carbon nanotube membrane structure 12 At this time, there is no obvious interface between the protein layer 14 and the carbon nanotube film structure 12. Generally, when the thickness of the carbon nanotube film structure 12 is thin, for example, when the thickness of the carbon nanotube film structure 2 is less than or equal to 3 μm, the protein in the protein layer is easily penetrated into the The carbon nanotube membrane structure 12 is described as being substantially coated with all of the carbon nanotubes in the carbon nanotube membrane structure 12. When the thickness of the carbon nanotube film structure 12 is relatively thick, for example, when the thickness of the carbon nanotube film structure 12 is greater than or equal to 3 μm, the protein in the protein (4) may penetrate into the protein. The inner surface of the carbon nanotube membrane structure 12 is covered with the carbon nanotube membrane structure _ network 20 carbon nanotubes. In the present invention, the protein in the protein layer 14 substantially covers all of the carbon nanotubes in the carbon nanotube membrane structure 12. [0046] The neural network 20 is disposed on a surface of the protein layer u away from the carbon nanotube film structure 12. When the nerve graft m includes only one of the carbon nanotubes 12 and the surface of the carbon nanotube 12-hydrophobic substrate u is provided with a protein layer 14, the nerve graft 100 includes only A neural network 2G is disposed on the surface of the protein layer. When the nerve graft (four) 〇 includes two nano carbon cut structures 12 respectively disposed on the two surfaces of the hydrophobic substrate 11 m + carbon " 099143857 Form No. A0101 Page 20 / Total 4 page 0992075912-0 201224149 [0047 ] Ο [0048]
G 結構12遠離疏水性基底11的表面均設置有—個蛋白質層 U時’所述神經移植體1GQ可包括㈣神經網賴分別設 置在所述兩個蛋白質層14的表面’亦可僅包括一個神經 網路20設置在其中-個蛋白質層14的表面。在本實施例 中，所述神經移植體剛僅包括—個奈米碳管膜結構12、 -個設置在所述奈米碳管膜結構12表面的蛋白質層⑷ 及一個*又在所述蛋白質層14表面的神經網路。 可以理解，為提高所述生物移植體1〇〇的抑菌性，提高該 神經移植體100的壽命，所述神經移植體1〇〇還可進一步 包括一多聚賴氨酸層設置在所述神經網路2〇與所述蛋白 質層14之間。 請參閲圖6，所述神經網路2〇包括複數神經細胞22及自所 述複數神經細胞22延伸出來的複數神經突起24。每一個 神經細胞22延伸出來的神經突起24的傭‘不限，只要能 夠使所述複數神經細胞22之間建立起生物連接使所述複 數神經細胞22能夠相互溝通即可。譬如，其中一個神經 細胞22可延伸出複數神經突起24或不延伸出任何神經突 起24。 [0049] 099143857 本發明中的神經移植體1QG，具有修衫物體中神經系統 中的神經網糊設置在該培育層1G表面。而所述_層 1〇中的疏水性基底11具有較好的，具有不含金屬、^性 佳、不易脑及延祕良好等優‘?卜所述培育㈣中的 奈米碳管膜結構12為基本由奈米碳管組成的自支揮社構 ，具有不含金屬、彈性佳、不易腐姓、延展性良好及1 密度等優點。故，該培育層1G可隨同該由複數神 表單編號A0101 第21頁/共4〇頁 、 巧 0992075912-0 201224149 24連接的複數神經細胞22 一起植入到生物體中，用於修 復生物體中受損的神經系統，且可根據生物體中神經系 統的創傷面積對所述神經移植體1〇〇進行裁剪或拉伸。 [0050] [0051] [0052] [0053] [0054] [0055] 以下將結合附圖並以具體實施例方式詳細說明本發明的 神經移植體的製備方法及神經移植體。 本發明提供一種神經移植體的製備方法，其包括如下步 驟： S210，提供一矽膠基底。 所述矽膠基底的尺寸與厚度可根據實際需秦而確定。譬 如，如果所需神經移植體的面積為3平方釐米，則所述矽 膠基底的面積可大於等於3平方釐米。所述矽膠基底不含 金屬，對生物體基本無毒性。‘ S220，將一奈米碳管膜結構鋪設在所述矽膠基底的一表 面0 為使所述奈Μ管膜結構與所述㈣基底表面結合更緊 密’可對所述奈来碳營膜輯進行有機溶劑處理。具體 地’可在設置在所述娜基底表面的奈米碳管膜結構覆 蓋或者滴上容易揮發的溶劑，如有機溶劑 ，再使所述溶 劑揮發，從而可減小料米碳㈣結構的比表面及增加 該奈米碳管膜結構與所述謂基底的附著力。所述奈米 碳管膜結構包括複數層冑㈣ ，相鄰的奈米碳管 拉膜之間的奈米碳管的延伸方向具有—交叉角度。請參 閱圖7及圖8 ’優選地，所述交叉角度大致等於90度。 099143857 表單編號A0101 第22 頁/共40頁 0992075912-0 201224149 [0056] S230，將鋪設有奈米碳管膜結構的矽膠基底浸泡在—蛋 白質溶液中。 [0057] 由於所述奈米碳管膜結構設置在所述矽膠基底表面，當 所述奈米碳管膜結構連同石夕膠基底一起浸泡到所述純蛋 白質中時，所述奈米碳管膜結構受液體表面張力影響而 產生破損的概率將大為降低。所述蛋白質溶液純牛灰清 溶液。請參見圖9，當所述奈米碳管膜結構從所述蛋白質 溶液中浸泡1. 5個小時左右，所述奈米碳管膜結構中的大 0 部分奈米碳管表面可浸潤有蛋白賀溶液。 [0058] S240，從所述蛋白質溶液中麥出所述鋪設有奈米碳管膜 結構的矽膠基底，在120攝民度下進行高溫減菌處理，形 成一培育層。 二’ [0059] 將所述鋪設有奈米碳管膜結構的矽膠基底從所述蛋白質 溶液中取出後’可在一乾燥箱中進行加熱滅菌處理。所 述乾燥箱的滅菌溫度大致在120度左右，減菌處理後後， 所述蛋白質溶液中的蛋白質基本©化，在所述奈米碳管 膜結構表面形成一蛋白質層，從而形成所述培育層。 [0060] S250，將所述培育層浸泡在一聚賴氨酸溶液中。 [0061] 所述多聚賴氨酸溶液中的多聚賴氨酸的濃度大致為2〇微 克每毫升。通過浸泡，所述培育層表面附著有多聚賴氨 酸，並提供一個水性環境。 [0062] S260 ’在所述浸泡’後的培育層滴加一神經細胞液直到該 神經細胞液覆蓋該培育層。 099143857 表單編號A0101 第23頁/共4〇頁 0992075912-0 201224149 [0063] [0064] 由於所述培育層中具有 —μ Χ膠基底，該培育層可直接設置 在一谷益如培養皿中 , 逋過所述矽膠基底係所述奈米碳 官膜結構不與所述” Μ表面接觸。 270培月所述附考在所述培育層的複數神經細胞，使 該複數神經細胞生長㈣域接麵述複數神 經細胞之間，“在所述培育層形成—神經網路。 闺所述神㈣胞㈣育環料普通的室㈣境，培育時間 可根據實際f求㈣。故，在步職6Q㈣境下，請參 閱圖10，保持錢條件不變，在㈣環境培養15天左右 ，即可使所述神經細胞分化出多個神經突起。所述神經 細胞生長時’所述蛋白質如牛血清，能夠提供供所述神 '二細胞生長的生長因數。所述多.個神經細胞上的多個神 經犬起相互連接後，形成所述神經網路及移植體請參 閱圖U及圖12，為所述神經移植體未經染色的掃描電鏡 照片及經過染色後的透射電鏡照片。從上述投射電鏡照 片可以清晰看出，所述神經移植體中的多個神經細胞通 過神經突起連接在一起。同時，如_n所示，部分神經 細胞雖然延伸出多個神經突起，但並未通過該多個神經 突起與其他神經細胞連接在一起，但這並不影響該神經 移植體在整體上具有生物活性的性質。 [0066]需要指出的係，由於所述培育層中的矽膠基底將所述奈 米碳管膜結構所述容器隔開，而該矽膠基底本身具有疏 水性。故，所述神經細胞將僅吸附在設置在所述奈米石炭 管骐結構上蛋白質層表面且在該表面生長，而不會在所 述矽膠基底表面或容器的内表面上生長。 0992075912-0 Γ 表單編號A0101 第24頁/共40頁 201224149 [0067] 綜上所述，本發明確已符合發明專利之要件，遂依法提 出專利申請。惟，以上所述者僅為本發明之較佳實施例 ，自不能以此限制本案之申請專利範圍。舉凡習知本案 技藝之人士援依本發明之精神所作之等效修飾或變化， 皆應涵蓋於以下申請專利範圍内。 【圖式簡單說明】 [0068] 圖1為本發明實施例所提供的一神經移植體的製備方法的 流程示意圖。The G structure 12 is disposed away from the surface of the hydrophobic substrate 11 with a protein layer U. The nerve graft 1GQ may include (4) the neural network lie disposed on the surface of the two protein layers 14 respectively. The neural network 20 is disposed on the surface of the protein layer 14 therein. In this embodiment, the nerve graft has just included a carbon nanotube membrane structure 12, a protein layer (4) disposed on the surface of the carbon nanotube membrane structure 12, and a protein in the protein. The neural network on the surface of layer 14. It can be understood that, in order to improve the bacteriostasis of the biological graft, and to increase the life of the nerve graft 100, the nerve graft may further include a poly-lysine layer disposed in the The neural network 2 is interposed between the protein layer 14. Referring to Figure 6, the neural network 2 includes a plurality of neural cells 22 and a plurality of neurites 24 extending from the plurality of neural cells 22. The neurites 24 from which each of the nerve cells 22 extends are not limited as long as the biological connection between the plurality of nerve cells 22 can be established to enable the plurality of nerve cells 22 to communicate with each other. For example, one of the nerve cells 22 may extend beyond the plurality of neurites 24 or may not extend any neurites24. 099143857 The nerve graft 1QG of the present invention has a nerve net paste in a nervous system in a dressing object disposed on the surface of the growth layer 1G. The hydrophobic substrate 11 in the layer 1 has good properties, and has a metal-free, good-quality, non-brain-like, and good-precision, and the carbon nanotube membrane structure in the cultivation (4). 12 is a self-supporting structure consisting mainly of carbon nanotubes, which has the advantages of no metal, good elasticity, non-perishable surname, good ductility and 1 density. Therefore, the culture layer 1G can be implanted into the living body together with the plurality of nerve cells 22 connected by the plural number form A0101 page 21/4 pages, Qiao 0992075912-0 201224149 24 for repairing the living body. The damaged nervous system, and the nerve graft 1 can be cut or stretched according to the wound area of the nervous system in the living body. [0055] Hereinafter, a method for preparing a nerve graft of the present invention and a nerve graft will be described in detail with reference to the accompanying drawings and specific embodiments. The invention provides a preparation method of a nerve graft, which comprises the following steps: S210, providing a silicone substrate. The size and thickness of the silicone substrate can be determined according to actual needs. For example, if the area of the desired nerve graft is 3 square centimeters, the area of the silicone substrate may be greater than or equal to 3 square centimeters. The silicone substrate is metal free and substantially non-toxic to the organism. ' S220, laying a carbon nanotube film structure on a surface of the silicone substrate 0 to make the natrital film structure and the (4) substrate surface more tightly combined with the nai carbon film Perform organic solvent treatment. Specifically, the carbon nanotube film structure disposed on the surface of the substrate can be covered or floated with a solvent which is easily volatilized, such as an organic solvent, and then the solvent is volatilized, thereby reducing the ratio of the carbon (four) structure of the rice material. The surface and the adhesion of the carbon nanotube film structure to the said substrate. The carbon nanotube membrane structure comprises a plurality of layers (four), and the direction of extension of the carbon nanotubes between the adjacent carbon nanotube membranes has an angle of intersection. Please refer to Figures 7 and 8'. Preferably, the angle of intersection is substantially equal to 90 degrees. 099143857 Form No. A0101 Page 22 of 40 0992075912-0 201224149 [0056] S230, a silicone substrate having a carbon nanotube film structure is immersed in a protein solution. [0057] Since the carbon nanotube film structure is disposed on the surface of the tantalum substrate, the carbon nanotube film is immersed in the pure protein together with the base of the gelatin gum, the carbon nanotube The probability that the membrane structure is damaged by the surface tension of the liquid will be greatly reduced. The protein solution is a pure ash ash solution. Referring to FIG. 9, when the carbon nanotube membrane structure is immersed from the protein solution for about 1.5 hours, the surface of the large carbon nanotube membrane in the carbon nanotube membrane structure may be infiltrated with protein. Hehe solution. [0058] S240, removing the tantalum substrate with the carbon nanotube film structure from the protein solution, and performing high temperature sterilization treatment at 120 degrees Celsius to form a seed layer. [0059] The silicone substrate having the carbon nanotube film structure is taken out from the protein solution and then subjected to heat sterilization treatment in a dry box. The sterilization temperature of the drying oven is approximately 120 degrees. After the sterilizing treatment, the protein in the protein solution is substantially chemicalized, and a protein layer is formed on the surface of the carbon nanotube membrane structure to form the cultivation. Floor. [0060] S250, immersing the incubation layer in a poly-lysine solution. [0061] The concentration of polylysine in the polylysine solution is approximately 2 μg per ml. By soaking, the surface of the layer is attached with polylysine and provides an aqueous environment. [0062] S260 ' drops a nerve cell fluid on the incubation layer after the soaking' until the nerve cell fluid covers the incubation layer. 099143857 Form No. A0101 Page 23/Total 4 Page 0992075912-0 201224149 [0064] Since the incubation layer has a -μ silicone substrate, the cultivation layer can be directly disposed in a oyster dish, The silicone substrate is not in contact with the surface of the ruthenium. The 270-peth-period is attached to the plurality of nerve cells in the culturing layer, so that the plurality of nerve cells grow (four) Between plural nerve cells, "formed in the incubation layer - neural network.闺The god (four) cell (four) cultivating the ordinary room (four) environment, the cultivation time can be based on the actual f (four). Therefore, in the 6Q (4) step, please refer to Figure 10, keep the money conditions unchanged, and in the (4) environment culture for about 15 days, the nerve cells can be differentiated into multiple neurites. When the nerve cells are grown, the protein, such as bovine serum, is capable of providing a growth factor for the growth of the divine cells. After the plurality of nerve dogs on the plurality of nerve cells are connected to each other, the neural network and the transplant body are formed. Referring to FIG. 9 and FIG. 12, the scanning electron micrographs and the images of the nerve grafts are not stained. Transmission electron micrographs after staining. As is clear from the above-described projection electron micrograph, a plurality of nerve cells in the nerve graft are connected by neurites. At the same time, as shown by _n, although some nerve cells extend out of multiple neurites, they are not connected to other nerve cells through the multiple neurites, but this does not affect the whole body of the nerve graft. The nature of the activity. [0066] It is noted that the tantalum substrate itself is hydrophobic due to the silicone substrate in the incubation layer separating the container of the carbon nanotube membrane structure. Therefore, the nerve cells will only adsorb on and grow on the surface of the protein layer disposed on the structure of the carbon nanotubes without growing on the surface of the silicone substrate or the inner surface of the container. 0992075912-0 表单 Form No. A0101 Page 24 of 40 201224149 [0067] In summary, the present invention has indeed met the requirements of the invention patent and has filed a patent application according to law. However, the above description is only a preferred embodiment of the present invention, and it is not possible to limit the scope of the patent application of the present invention. Equivalent modifications or variations made by those skilled in the art in light of the spirit of the invention are intended to be included within the scope of the following claims. BRIEF DESCRIPTION OF THE DRAWINGS [0068] FIG. 1 is a schematic flow chart of a method for preparing a nerve graft according to an embodiment of the present invention.
[0069] 圖2為一奈米碳管絮化膜的掃描電鏡照片。 [0070] 圖3為一奈米碳管碾壓膜的掃描電鏡照片。 [0071] 圖4為一奈米碳管拉膜的掃描電鏡照片。 [0072] 圖5為本發明實施例所提供的神經移植韙的側視示意圖。 [0073] 圖6為本發明實施例所提供的神經移植體的俯視示意圖。 [0074] 圖7為本發明實施例所提供的奈米碳管膜結構的掃描電鏡 照片。 [0075] 圖8為本發明實施例所提供的奈米碳管膜結構的透射電鏡 照片。 [0076] 圖9為本發明實施例所提供的培育層的透射電鏡照片。 [0077] 圖1 0為本發明實施例所提供的種植在所述培育層上的神 經細胞分化出多個神經突起時的掃描電鏡照片。 [0078] 圖11為本發明實施例所提供的未經染色的神經移植體的 掃描電鏡照片。 099143857 表單編號A0101 第25頁/共40頁 0992075912-0 201224149 [0079] 圖12為本發明實施例所提供的神經移植體染色後的掃描 電鏡照片。 【主要元件符號說明】 [0080] 神經移植體：100 [0081] 培育層：10 [0082] 疏水性基底：11 [0083] 奈米碳管膜結構：1 2 [0084] 蛋白質層：1 4 [0085] 神經網路：20 [0086] 神經細胞：22 [0087] 神經突起：24 0992075912-0 099143857 表單編號A0101 第26頁/共40頁2 is a scanning electron micrograph of a carbon nanotube flocculation membrane. 3 is a scanning electron micrograph of a carbon nanotube rolled film. 4 is a scanning electron micrograph of a carbon nanotube film. 5 is a side view of a nerve graft according to an embodiment of the present invention. 6 is a schematic top view of a nerve graft according to an embodiment of the present invention. 7 is a scanning electron micrograph of a carbon nanotube film structure provided by an embodiment of the present invention. 8 is a transmission electron micrograph of a carbon nanotube film structure provided by an embodiment of the present invention. 9 is a transmission electron micrograph of a cultivating layer provided by an embodiment of the present invention. 10 is a scanning electron micrograph of a neural cell implanted on the growth layer to differentiate into a plurality of neurites according to an embodiment of the present invention. 11 is a scanning electron micrograph of an unstained nerve graft according to an embodiment of the present invention. 099143857 Form No. A0101 Page 25 of 40 0992075912-0 201224149 [0079] FIG. 12 is a scanning electron micrograph of a nerve graft after staining according to an embodiment of the present invention. [Explanation of main component symbols] [0080] Nerve graft: 100 [0081] Cultivated layer: 10 [0082] Hydrophobic substrate: 11 [0083] Carbon nanotube membrane structure: 1 2 [0084] Protein layer: 1 4 [ 0085] Neural Network: 20 [0086] Nerve Cells: 22 [0087] Nerve Protrusion: 24 0992075912-0 099143857 Form Number A0101 Page 26 of 40