201224146 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的製備方法，尤其係涉及一 種可供生物體移植的神經移植體的製備方法。 【先前技術】 [0002] 〇 〇 [0003] 099143853 神經系統主要由神經元（neurons)以及神經膠質細胞 (neuron glial 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 經系統中，係由神經元來執行接收刺激、通過傳導並輪 出神經遞質（neuron transmit ter)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元物理性支援 、營養提供以及調節溝通訊息迷度等功能。畚一神經元 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轴 突（axon)與樹突（dendrite)兩種。一般來說，刺激由 樹突接收並將衝動傳向胞體，衝動經過軸突傳導至轴★ 末端，並釋放傳導物質來觸動其俋‘胞。 由於神經系統扮演生物鱧内各組織與器官之間的協調作 用，其重要性不言可喻。先前，因神經系統受損而導致 的神經缺損係臨床常見的致殘性疾病。其中，通過植入 神經移植體來修復受損的神經系統，係神經外科手術用 來修復因各種情況引起的神經系統損傷的一種重要手段 。先前的神經移植體通橋接，，在神㈣統受損部 位兩端的神經管，該神經管由生物降解材料製成的管狀 表單編號A0101 第3頁/共41頁 ^92075908-0 201224146 結構。神經系統受損部位一端的神經元沿所述神經管内 壁生長出神經突起以到達所述神經系統受損部位的另一 端。 [0004]通常，需要通過植入所述神經管的方式進行修復的的受 損部位的長度較長，而所述神經突起的生長過程非常緩 杈，故，利用所述神經管來修復受損的神經系統所需的 修復時間較長。 【發明内容】 剛有鑒於此，提供一種神經移植體的製備方法實為必要， 由該製備方法製備的神經移植體能夠減少受損的神經系 統的修復時間。 [〇〇〇6] 一種神經移植體的製備方法，其包括：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疏水性基底 的一表面，所述蛋白質層設i在該奈米碳管膜結構遠離 所述疏水性基底的表面；在該蛋白質層遽離所述疏水性 基底的表面種植複數神經細胞；以及培育該複數神經細 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。 [0007] 099143853 一種神經移植體的製備方法，其包括如下步驟：提供一 mm找表面舖設—奈米碳管 膜結構；在該奈米碳管膜結構遠離所•水性基底的表 面形成〜蛋白質層從而形成-培育層；在該蛋白質声遠 離所㈣水性基底的表面種植複數神^胞；以及培育 该複數神經細胞直到該複數神經細 表單編第4頁/共41頁胞生長出複數神經突 201224146 [0008] Ο ❹ [0009] [0010] [0011] 起連接在所述複數神經細胞之間形成一神經網路β 相較於先前技術，所述神經移植體的製備方法通過在該 奈米碳管膜結構表面設置蛋白質層形成培育層，並在所 述培育層的表面形成所述神經網路。所述奈米碳管膜結 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信號傳遞能力。 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 神經元的距離非常近，故可通過直接鏠合所述神經植入 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】 請參閱圖1，本發明提供一種神經移植體的製備方法，其 包括： 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、结構設 置在所述矽膠基底，所述蛋白質層設置在該奈米破管滕 結構表面； S20，在該蛋白質層表面種植複數神經細胞；以及 099143853 表單編號Α0101 0992075908-0 第5頁/共41頁 201224146 剛S3G，培育該複數神經細胞直㈣減神經細胞生長出複 數神經突起連接在所述複數神經細胞之間形成—神經網 路。 _纟所述步驟S1Q中，所述奈米碳管膜結構由複數奈米碳管 所^«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 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 保持自身特定的形狀〇隹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 米礙管膜結構具有特定的形秦，形成一自支擇結構。所 述奈米碳管膜結構為自支撐結構時，該奈米碳管膜結構 可為由至少一奈米碳管膜形成的膜狀結構，當所述奈米 碳管膜結構包括複數奈米碳管膜時，該複數奈米碳管膜 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 之間通過凡知·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，便於裁剪和拉伸。 [0014]請參閱圖2，所述奈米碳管膜可為一奈米碳管絮化膜該 奈米碳管絮化膜為將一奈米碳管原料絮化處理獲得的— 自支推的奈米碳管獏。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 099143853 表單編號A0101201224146 VI. Description of the Invention: [Technical Field] [0001] The present invention relates to a method for preparing a nerve graft, and more particularly to a method for preparing a nerve graft for transplantation of an organism. [Prior Art] [0002] 〇〇[0003] 099143853 The nervous system is mainly composed of neurons (neurons) and neuron glial cells, which form a complex and specific communication domain to establish links with other tissues or organs. For functional coordination. In the nervous system, neurons receive stress, conduct and rotate neurotransmitters (neuron transmit ter) for tissue or organ communication, while glial cells perform neuronal physical support, nutrient supply, and Adjust the functionality of communication messages and other features. The neuron includes two parts, the cell body and the neurite. The neurites extend from the cell body and grow toward other neurons or other cells (eg, muscle cells), in which the neurites Divided into axon (axon) and dendrites (dendrite). In general, the stimulus is received by the dendrites and transmits impulses to the cell body, and the impulse is transmitted through the axons to the end of the axis, and the conductive material is released to touch the cells. Since the nervous system plays a coordinating role between tissues and organs within the biopterin, its importance is self-evident. Previously, neurological deficits due to impaired nervous system were clinically common disabling diseases. Among them, the implantation of a nerve graft to repair a damaged nervous system is an important means for neurosurgery to repair nervous system damage caused by various conditions. The previous nerve graft was bridged, and the neural tube at both ends of the damaged part of the god (four) system was made of biodegradable material. Form No. A0101 Page 3 of 41 ^92075908-0 201224146 Structure. Neurons at one end of the damaged portion of the nervous system grow neurites along the inner wall of the neural tube to reach the other end of the damaged portion of the nervous system. [0004] Generally, the length of the damaged portion that needs to be repaired by implanting the neural tube is long, and the growth process of the neurite is very slow, so the nerve tube is used to repair the damage. The nervous system requires a longer repair time. SUMMARY OF THE INVENTION In view of the above, it is necessary to provide a method for preparing a nerve graft, and the nerve graft prepared by the preparation method can reduce the repair time of the damaged nervous system. [6] A method for preparing a nerve graft, comprising: providing a cultivating layer comprising a hydrophobic substrate, a carbon nanotube membrane structure, and a protein layer, the carbon nanotube a membrane structure disposed on a surface of the hydrophobic substrate, the protein layer being disposed on a surface of the carbon nanotube membrane structure away from the hydrophobic substrate; implanted on a surface of the protein layer that is separated from the hydrophobic substrate a plurality of nerve cells; and 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. [0007] 099143853 A method for preparing a nerve graft, comprising the steps of: providing a mm surface-laid-nanocarbon tube membrane structure; forming a ~protein layer on the surface of the carbon nanotube membrane structure away from the aqueous substrate Thereby forming a layer of cultivation; planting a plurality of cells in the surface of the protein sound away from the surface of the (4) aqueous substrate; and cultivating the plurality of nerve cells until the plurality of nerves form a page 4th/41 pages of cells growing a plurality of neurites 201224146 [0008] [0011] [0011] a neural network is formed between the plurality of neural cells to form a neural network. Compared to the prior art, the preparation method of the nerve graft is performed on the nanocarbon. The surface of the tubular membrane structure is provided with a protein layer to form a growth layer, and the neural network is formed on the surface of the cultivation layer. The carbon nanotube membrane structure and the hydrophobic substrate have the advantages of good elasticity and good ductility. Therefore, the nerve graft can be cut and stretched according to the shape and size of the damaged portion of the damaged nervous system. Implant the damaged area. The neural network has biological activity and signaling capabilities' so that the neural graft including the neural network also has biological activity and signal transmission capabilities. When the nerve graft is implanted into the damaged part of the living body, since the neuron in the nerve implant is very close to the neurons at the two ends or the edge of the damaged part, it can be directly The manner of combining the neurons in the nerve implant with the neurons at the edge of the damaged site enables the two ends of the damaged portion to establish a signal transmission capability, complete the nerve repair of the damaged portion, thereby saving the nerve The growth time of the protrusions reduces the repair time of the damaged nervous system. [Embodiment] Please refer to FIG. 1 , the present invention provides a method for preparing a nerve graft, comprising: S10 , providing a cultivating layer comprising a hydrophobic substrate, a carbon nanotube membrane structure and a protein a layer, the carbon nanotube film, a structure disposed on the silicone substrate, the protein layer disposed on the surface of the nanotube structure; S20, implanting a plurality of nerve cells on the surface of the protein layer; and 099143853 Form No. Α0101 0992075908-0 Page 5 of 41201224146 Just S3G, cultivating the plural nerve cells straight (four) reducing nerve cells grow a plurality of neurites connected to form a neural network between the plurality of nerve cells. In the step S1Q, the carbon nanotube film structure may extend substantially parallel to the surface of the carbon nanotube film structure by a plurality of carbon nanotubes. Preferably, the plurality of carbon nanotubes are connected by a Van der Waals force to form a self-supporting structure. The so-called "self-supporting" means that the carbon nanotube membrane structure can maintain its own specific shape without being disposed on the surface of a substrate, and a large number of carbon nanotubes pass through the self-supporting carbon nanotube membrane structure. The wattages are attracted to each other, so that the nano-membrane structure has a specific shape and forms a self-selective structure. When the carbon nanotube membrane structure is a self-supporting structure, the carbon nanotube membrane structure may be a membrane-like structure formed by at least one carbon nanotube membrane, and when the carbon nanotube membrane structure comprises a plurality of nanometer membrane structures In the case of the carbon nanotube film, the plurality of carbon nanotube films 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 a common gas force, the carbon nanotube film structure has the advantages of good elasticity, good ductility and low density. Easy to cut and stretch. [0014] Please refer to FIG. 2, the carbon nanotube film may be a carbon nanotube flocculation membrane, and the carbon nanotube flocculation membrane is obtained by flocculation of a carbon nanotube raw material - self-supporting The carbon nanotubes of the carbon nanotubes. 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 attracted to each other by van der Waals force. 099143853 Form No. A0101
第6頁/共41 I 0992075908-0 201224146 ，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。 [0015] 所述奈米碳管膜可為一奈米碳管碾壓膜，該奈米碳管碾 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召，其中，冷大於 等於0度且小於等於15度，該夾角召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 099143853 表單編號A0101 第7頁/共41頁 0992075908-0 201224146 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。 [0016] 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 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。 [0017] 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。 [0018] 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 099143853 表單編號A0101 第8頁/共41頁 0992075908-0 201224146 伸方向上相鄰的奈米碳管通過凡得瓦力首尾相連。當然 • 斤述不米碳管拉膜中存在少數偏離該延伸方向的奈米 反S這些奈米碳管不會對奈米碳管拉膜中大多數奈米 奴官的整體取向排列構成明顯影響。所述自支樓為奈米 破S拉膜不需要大面積的載體支標，而只要相對兩邊提 供支擇力即能整體上懸空而保持自身膜狀狀態，即將該 不米奴官拉膜置於（或固定於）間隔一定距離設置的兩 個支擇體上時，位於兩個支㈣之間的奈米碳管拉膜能 ο 持自身雜« n自麟主要通過奈米碳 Β拉膜中存在連續㈣過凡得瓦力連延伸排列的 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 適田的f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 〇 連續且疋向排列的奈米❹>5段該複數奈求碳管片段 通過凡付瓦力首尾相連。每—奈米碳管片段由複數相互 平行的奈来碳管組成。該奈来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以上。 闺*該Μ碳管赌構包括複數Μ碳錄糾，所述複 數奈米碳管拉膜層疊設置形成-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来碳管膜的層數小於或 099143853 表單編號Α0101 第9頁/共41頁 0992075908-0 201224146 等於1 〇層，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α， 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 ，相鄰的奈米碳管膜中的奈米碳管之間的交叉角度α大 致等於90度，即，相鄰奈米碳管拉膜中的奈米碳管的延 伸方向大致平行。 [0020] 所述疏水性基底用於承載該所述奈米碳管膜結構及蛋白 質層。所述疏水性基底具有疏水性及良好的柔韌性。所 述疏水性基底可由矽膠製成，或表面塗敷有矽膠。所述 疏水性基底的形狀與厚度可根據所述奈米碳管膜結構的 形狀與厚度設計。譬如，所述疏水性基底表面的面積及 形狀可大致與所述奈米碳管膜結構的面積及形狀大致相 當。可以理解，當所述奈米碳管膜結構的厚度較薄時， 該奈米碳管膜結構具有較小機械強度及具有較大的比表 面積，故，該奈米碳管膜結構溶液受外力產生破損或容 易粘附在其他親水性物體上。將該奈米碳管膜結構設置 在所述疏水性基底表面形成一生物基底時，該生物基底 的機械強度較所述奈米碳管膜結構大，從而使該奈米碳 管膜結構更難受外來作用而產生破損，同時便於移動及 099143853 表單編號Α0101 第10頁/共41頁 0992075908-0 201224146 防止該奈米碳管膜結構粘附在親水性物體上。 [0021] Ο 所述蛋白質層設置在所述奈米碳管膜結構的表面，用於 使所述奈米碳管層具有親水性及生物相容性，從而使得 所述培育層能夠為所述神經細胞的種植及生長提供一個 合適的環境。具體地，所述蛋白質層設置在所述奈米碳 管膜結構遠離所述疏水性基底的表面。優選地，所述蛋 白質層中的蛋白質（protein)為可溶性蛋白質。所述蛋 白質可選自纖維狀蛋白質、血漿蛋白質及酶蛋白質等。 在本實施例中，所述蛋白質為鳴乳動物的血清，如豬血 清、牛血清或人血清辦述血清不僅能夠為所述神經細 胞的種植及生長提供一個合適的環境’還能夠在所述神 經細胞生長時，為所述神經細胞提供生長因數。 [0022] Ο 所述培育層的製備方法不限，只要能夠像奎白質層與所 述奈米碳管膜結構混合在一起即可。譬如，可通過將所 述奈米碳管膜結構浸泡在一蛋白質溶液中，使所述蛋白 質溶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白質層。亦可將所述蛋白質溶液 噴塗在所述奈米碳管膜結構遠離所述疏水性基底的表面 ，使所述蛋白質層設置在該表面。還可將蛋白質溶液滴 在所述奈米碳管膜結構遠離所述疏水性基底的表面，再 採用甩膜的方式使所述蛋白質層設置在該表面°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 media) ’所述生物媒介的種類 不限，可根據蛋白質的種類的不同而調製。通常，所述 099143853 表單編號A0101 第11頁/共41頁 0992075908-0 201224146 蛋白質溶液中的蛋白質的濃度大於等於50%小於等於100% 。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 〇 [0023] 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 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： [0024] 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： [0025] S11，提供所述疏水性基底； [0026] S12，將所述奈米碳管膜結構設置在所述疏水性基底表面 099143853 表單編號A0101 第12頁/共41頁 0992075908-0 201224146 [0027] [0028] [0029] Ο [0030] 〇 [0031] 099143853Page 6 of 41 I 0992075908-0 201224146 , forming 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 500 nm. The gap or micropores can increase the specific surface area of the carbon nanotube membrane and infiltrate more protein. [0015] The carbon nanotube film may be a carbon nanotube rolled film, and the carbon nanotube film is a self-supporting nano carbon obtained by rolling a carbon nanotube array. Tube membrane. 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 with the surface of the growth substrate forming the carbon nanotube array, wherein the cold is greater than or equal to 0 degrees and less than or equal to 15 degrees, and the angle is called and applied. The pressure on the carbon nanotube array is related. The larger the pressure, the smaller the angle. 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; please 099143853 Form No. A0101 Page 7 / Total 41 Page 0992075908-0 201224146 Referring to Figure 3, when rolling in different directions, the carbon nanotubes are arranged in different directions. When rolled in the same direction, the carbon nanotubes are arranged in a preferred orientation along a fixed direction. The length of the carbon nanotubes in the carbon nanotube rolled film is greater than 50 microns. [0016] 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 smaller the applied pressure, the larger the thickness of the prepared carbon nanotube rolled film; on the contrary, the smaller the height of the carbon nanotube array, the more the applied pressure Large, 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. [0017] 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 carbon nanotubes are preferably aligned 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. [0018] Further, most of the carbon nanotubes in the carbon nanotube film are connected end to end by van der Waals force. Specifically, in the carbon nanotube film, most of the carbon nanotubes in the same direction extend substantially in the same direction as each of the carbon nanotubes in the extension 099143853 Form No. A0101 Page 8 / Total 41 Page 0992075908-0 201224146 The adjacent carbon nanotubes in the direction are connected end to end by van der Waals force. Of course, there are a few nano-carbon nanotubes in the film that are deviated from the direction of extension. These carbon nanotubes do not significantly affect the overall orientation of most of the nano slaves in the carbon nanotube film. . The self-supporting building does not need a large-area carrier support for the nano-broken S-film, but can maintain its own membranous state as long as it provides a supporting force on both sides, that is, the non-nano slave film is placed. When (or fixed to) two spacers arranged at a certain distance apart, the carbon nanotube film located between the two branches (four) can hold its own impurity. There is a continuous (four) continuous realization of the van der Waals and the arrangement of the carbon nanotubes. Specifically, the majority of the carbon nanotubes extending in the same direction in the carbon nanotube film are not absolutely linear, and may be arranged in the direction of the extension direction, and may be appropriately deviated. Extend the 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 〇 continuous and 疋-aligned nano ❹> 5 segments of the plurality of carbon tube segments connected end to end by a wattage force. Each of the carbon nanotube segments consists of a plurality of carbon nanotubes that are parallel to each other. The carbon nanotube segments have an arbitrary length uniformity and shape. The carbon nanotube film has good light transmittance and the visible light transmittance can reach above.闺* The carbon nanotubes include a plurality of carbon recordings, and 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 number of layers of the carbon nanotube film that is less than or 099143853. Form number Α0101 page 9/total 41 page 0992075908-0 201224146 equals 1 〇 layer, thereby making the carbon nanotubes per unit area The small amount keeps the Raman light intensity of the carbon nanotube itself 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 intersection angle α between the α 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 nanotube film laminated, and the intersection angle α between the carbon nanotubes in the adjacent carbon nanotube film is substantially equal to 90 degrees. That is, the extending directions of the carbon nanotubes in the adjacent carbon nanotube film are substantially parallel. [0020] The hydrophobic substrate is used to carry the carbon nanotube membrane structure and the protein layer. The hydrophobic substrate has hydrophobicity and good flexibility. The hydrophobic substrate may be made of silicone or coated with silicone. The shape and thickness of the hydrophobic substrate can be designed according to the shape and thickness of the carbon nanotube film structure. For example, the area and shape of the surface of the hydrophobic substrate may be substantially equivalent to the area and shape of the carbon nanotube film structure. It can be understood that when the thickness of the carbon nanotube film structure is thin, the carbon nanotube film structure has small mechanical strength and a large specific surface area, so the carbon nanotube membrane structure solution is subjected to an external force. It is damaged or easily adheres to other hydrophilic objects. When the carbon nanotube film structure is disposed on the surface of the hydrophobic substrate to form a biological substrate, the mechanical strength of the biological substrate is larger than that of the carbon nanotube film structure, thereby making the structure of the carbon nanotube film more uncomfortable. Damage caused by external action, and easy to move and 099143853 Form No. 1010101 Page 10 / Total 41 Page 0992075908-0 201224146 Prevent the carbon nanotube membrane structure from adhering to hydrophilic objects. [0021] Ο the protein layer is disposed on a surface of the carbon nanotube film structure for imparting hydrophilicity and biocompatibility to the carbon nanotube layer, thereby enabling the seed layer to be The cultivation and growth of nerve cells provides a suitable environment. Specifically, the protein layer is disposed on a surface of the carbon nanotube film structure away from the hydrophobic substrate. Preferably, the protein in the protein layer is a soluble protein. The protein may be selected from the group consisting of fibrous proteins, plasma proteins, and enzyme proteins. In this embodiment, the protein is a serum of a whey animal, such as porcine serum, bovine serum or human serum, which can not only provide a suitable environment for the growth and growth of the nerve cells. When the nerve cells grow, they provide a growth factor for the nerve cells. [0022] The preparation method of the cultivating layer is not limited as long as it can be mixed with the quinolin layer and the carbon nanotube film structure. For example, the protein solution can be infiltrated into the carbon nanotube membrane structure by immersing the carbon nanotube membrane structure in a protein solution: thereby allowing proteins in the protein solution to adhere to the na[beta] The carbon nanotube film structure forms a protein layer away from the surface of the hydrophobic substrate. The protein solution may also be sprayed onto the surface of the carbon nanotube membrane structure remote from the hydrophobic substrate such that the protein layer is disposed on the surface. A protein solution may also be dropped on the surface of the carbon nanotube membrane structure away from the hydrophobic substrate, and the protein layer may be disposed on the surface by means of a ruthenium membrane. The protein solution is in addition to the protein. A biological media that dissolves the protein may also be included. 'The type of the biological medium is not limited and may be modulated depending on the type of protein. Generally, the 099143853 Form No. A0101 Page 11 of 41 0992075908-0 201224146 The concentration of the protein in the protein solution is 50% or more and 100% or less. In this embodiment, the protein concentration in the protein solution is 100%, that is, the protein solution is pure protein, and no solvent is dissolved. [0023] Since the carbon nanotube membrane structure includes a plurality of carbon nanotube membranes And a gap exists between the plurality of carbon nanotubes to form a plurality of micropores, and when the protein solution infiltrates the carbon nanotube membrane structure, the protein solution can penetrate into the inner surface of the carbon nanotube membrane structure to infiltrate The surface of a plurality of carbon nanotubes. Of course, in the cultivating layer, not all the surface of the carbon nanotubes can be infiltrated with a protein layer, but only a part of the carbon nanotubes located on the surface of the carbon nanotube membrane structure in which the nerve cells need to be cultured are required. Infiltrating with a protein solution, a protein layer can be formed on the surface of the carbon nanotube membrane structure, so that the carbon nanotube membrane structure has hydrophilicity and biocompatibility, so that the growth layer has a carrier for nerve cell growth. The function. 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: [0024] In the embodiment, the method for preparing the cultivating layer may further include the following steps: [0025] S11, providing the hydrophobic layer a substrate; [0026] S12, the carbon nanotube film structure is disposed on the surface of the hydrophobic substrate 099143853 Form No. A0101 Page 12 / Total 41 Page 0992075908-0 201224146 [0028] [0029] Ο [0030] 99[991] 099143853
Sl3 ’使所述奈米碳管膜結構浸潤有蛋白 質溶液；以及 14 ’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。 在步驟S11中，所述疏水性基底由矽膠製成，或表面塗敷 有夕夥° @膠為生物體常用的植人材料，對生物體無毒 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⑦料疏水性基底可錢植入人體。 在步驟SI 2 H朗述奈米碳管膜結構與職謂基底 表面結合更緊密，可對所絲米碳㈣結構進行有機溶 劑處理。具體地，可在設置在㈣㈣基絲面的奈米 妷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_發，從而可減，〗、該奈#碳管膜結構的 =表面及増加該奈《管膜結構與所^膠基底的附著 力0 中，朗述奈料㈣•一有蛋白質溶液 的方式不限，只要使蛋白質溶液中的 ^ t ^ , 蛋白質附著在奈米 膜、-4構表面形成一蛋白質層即可。 。譬如，可通過將 所述奈米碳管膜結構浸泡在所述蛋白 真容液中，實現蛋 白質溶液的浸潤。亦可通過在所诚太也 T水碳管膜結構喷塗 所述蛋白質溶液’實現蛋白質溶液的& 丄 欠祠。在本實施例 中，為實現蛋白質溶液的浸潤，選禮妝 伴将所述奈米碳管膜 結構浸泡在純蛋白質中。所述奈米磁故Sl3' infiltrates the carbon nanotube membrane structure with a protein solution; and 14' sterilizes the carbon nanotube membrane structure infiltrated with the protein solution to form the incubation layer. In step S11, the hydrophobic substrate is made of silicone rubber, or the surface is coated with Xi'anjia gel as a common implant material for living organisms, which is non-toxic to the organism and has good flexibility. Therefore, the 7-material hydrophobic substrate prepared or coated from the silicone substrate can be implanted into the human body. In the step SI 2 H, the carbon nanotube membrane structure is more closely combined with the surface of the precursor substrate, and the organic carbon solution can be treated on the silk carbon (four) structure. Specifically, the nanotube film structure disposed on the (four) (four) base surface may be covered or dripped with a solvent which is easily volatilized, such as an organic solvent, and then the solution may be reduced, thereby reducing, and the carbon tube The surface structure of the membrane structure and the addition of the Nai "the adhesion of the membrane structure to the substrate of the gelatinous material 0, the description of the material (4) • a method of protein solution is not limited, as long as the protein solution in the ^ t ^, protein It can be attached to the surface of the nano-film and form a protein layer. . For example, the infiltration of the protein solution can be achieved by immersing the carbon nanotube membrane structure in the protein solution. It is also possible to achieve a protein solution & 祠 祠 by spraying the protein solution in the T. In this embodiment, in order to achieve infiltration of the protein solution, the makeup makeup is accompanied by soaking the carbon nanotube membrane structure in pure protein. The nanometer
、s犋結構的浸泡時 間依奈米碳管膜結構的具體結構及I 句質溶液的具體組 表單編號A0101 第13頁/共41頁 0992075908-0 201224146 分而定，只要能使所述奈米碳管膜結構中的大部分奈米 碳管浸潤有蛋白質溶液即可。通常，所述奈米碳管膜結 構的浸泡時間在2小時以上。所述奈米碳管膜結構的浸泡 的環境不限，只要不使所述蛋白質變質即可。通常，所 述浸泡過程可在常溫、常壓環境下進行。 [0032] 當所述奈米碳管膜結構浸泡在所述蛋白質溶液中時，所 述蛋白質溶液可浸潤所述奈米碳管膜結構中的部分奈米 碳管，亦可浸潤所述奈米碳管膜結構中的全部奈米碳管 。通常地，當所述奈米碳管膜結構的厚度較薄時，譬如 所述奈米碳管膜結構的厚度小於等於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。 [0033] 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 099143853 表單編號A0101 第14頁/共41頁 0992075908-0 201224146 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。 0 [0034] 在步驟S10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，該 步驟S10還可進一步包括如下步驟：S15,在所述蛋白質 層遠離所述疏水性基底的表面形成一聚賴氨酸（Poly-D-lysine，PDL)層。具體地，可將所述培育層浸泡在 一聚賴氨酸溶液中，所述聚賴氨酸溶液中聚賴氨酸的濃 度大致為20微克每毫升。 〇 [0035] 在步驟S20中，所述神經細胞包括哺乳動物的神經細胞， 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 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，亦可採用將該培育層浸泡在所述含神經細胞的神經 細胞液中，只要使所述神經細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皿的一底面上，只要能 099143853 表單編號A0101 第15頁/共41頁 0992075908-0 201224146 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|的底面時，所述疏水性基底與所述底面 接觸，從而使得所述神經細胞僅分佈在蛋白質層表面。 [0036] 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 壓環境中即可生長。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 牛血清可提供生長因數，促進該神經細胞生長。當然， 亦可使該培育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，可模擬所述老鼠體内的生長環境。 [0037] 所述神經細胞在培育時，能夠長出複數神經突起（Neur-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，當所述培育層表面澂置有複數神經細胞 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 099143853 表單編號A0101 第16頁/共41頁 0992075908-0 201224146 Ο 起形成所述神經網路，使該複數神經細胞之間能相 通。相鄰的神經細胞的神經突起如果相遇，則會合為向 们神、i犬起。再譬如，當所述神經細胞在該培育層: 面以線狀或者陣列的方式排列時，且沿縱向方向的神t ㈣相距較近’而沿橫向方向的神經細胞相距較遠，： 時’所述核細胞所生長的神經細胞可基本沿所述縱向 方向（伸。為使所述神經細胞能夠在所述培育層表^ 線狀或者㈣的方式制，可選擇使所述奈米碳管^ 的奈米碳管基本沿同—方向延伸。通過培育，彼此 的神經細胞大多通過神賊起建立起連接，從而形 述神經網路°所述神經網路與所述培育.層-起形成所 神經移植體。 ^ [0038] Ο 所述移植體的製財法通過在簡雜基絲面設置奈 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φ種植複數相細胞，並使所述複數神經 細胞生長出複數神經突起連接起魏神經細I所述石夕 膝基底具有疏水性，^能使所述#經細胞制其上，即 不能提供供所料經細胞生長的環境，故，所述神經細 胞將僅在所述設置有蛋白質層表面生長。 [0039] 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xl5毫米以上，具體地，該奈来碳管膜結 構的長度可達3GG米以上，寬度可達〇.5米以上。所述疏 水性基底亦為1觀結構，其形狀與面射根據所述奈 米碳管膜結構的形狀與面積進行調整4該奈米碳管膜 099143853 表單編號Α0101 第17頁/共41頁 0992075908-0 201224146 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，可直接植入生物體。故，由所述奈米碳 管膜結構及疏水性基底做主要載體的神經移植體可根據 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 有生物活性及信號傳遞能力。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 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 進行直接缝合，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。 [0040] 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 099143853 表單編號A0101 第18頁/共41頁 0992075908-0 201224146 的改性並不會對實現本發财任何實質性貢獻，因為， 田所述蛋白貝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。 [0041] [0043] ❹ [0044] [0045] 本發月乂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。 請參閱圖5，所述神經移植體1〇〇粤括一培育層1〇及分佈 在該培月層1〇表面的一神經網路2〇。 所述培育層10包括一疏水性基底Η、一奈米碳管膜結構 12及一蛋白質層14。所述奈米碳管膜結構〗2設置在所述 疏水性基底1丨的一個或者相對的兩個表面。所述蛋白質 層14設置在所述奈米碳管膜結構12遠離所述疏水性基底 11的表面。在本實施例中，所述奈米碳管臈結構12僅設 置在所述疏水性基底11的一個奉面》 所述疏水性基底11可由矽膠製成，或表面塗敷有碎膠。 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石夕 膠的疏水性基底11可直接植入人體。 所述奈米碳管膜結構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一自支撐結構。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 099143853 表單編號Α0101 第19頁/共41頁 0992075908-0 201224146 中的奈米碳管絮化膜、圖3中的奈米碳管碾壓膜及圖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，相鄰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 具有一個交叉角度，優選地，所述交叉角度為90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0. 3微米小於60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。 [0046] 所述蛋白質層14為由可溶性蛋白質組成。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，只要能夠提供一個親水性環境即可。通常， 所述蛋白質層14的厚度為0. 3微米到2微米。在本實施例 中，所述蛋白質層的厚度大致為0. 5微米。在宏觀上，所 述蛋白質層14可選擇僅設置在所述奈米碳管膜結構12遠 離所述疏水性基底11的表面。在微觀上，所述蛋白質層 14中的蛋白質容易滲透到所述奈米碳管膜結構12的内部 ，並包覆所述奈米碳管膜結構12中的部分或者全部奈米 碳管，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，所述奈米碳管膜結構12的厚度 小於等於3微米時，所述蛋白質層14中的蛋白質容易滲透 到所述奈米碳管膜結構12的内部，並基本包覆所述奈米 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 099143853 表單編號A0101 第20頁/共41頁 0992075908-0 201224146 [0047] Ο ο [0048] 的厚度大於等於3微米時，所述蛋白質層14中的蛋白質雖 然亦可滲透到所述奈米碳管膜結構12内部，然通常僅包 覆所述奈米碳管膜結構12靠近所述神經網路2〇的奈米碳 管。在本實施例中，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。 所述神經網路20設置在所述蛋白質層14遠離所述奈米碳 官膜結構12的一個表面。當所述神經移植體1〇〇僅包括一 個奈米碳官膜結構12且该奈米碳管膜結構12遠離疏水性 基底11的表面設置有一個蛋白質層14時，所述神經移植 體1 0 0僅包括一佩神經爾路20設置在所述蛋白質層丨4表面 。當所述神經移植體100包括兩個奈米碳管膜結構丨2分別 設置在該疏水性基底Π的兩個表面，且每一条米碳管膜 結構1 2遠離疏水性基底11的表面均設置有一個蛋白質詹 14時，所述神經移植體100可包括兩偭神經網路2〇分別設 置在所述兩個蛋白質層14的表面s，亦可僅包括一個神經 網路20設置在其中一個蛋白質層14的表面。在本實施例 中，所述神經移植體10 0僅包括一個奈米碳管膜結構12、 一個設置在所述奈米碳管膜結構12表面的蛋白質層14、 及一個設在所述蛋白質層14表面的神經網路20。 可以理解，為提高所述生物移植體100的抑菌性，提高該 神經移植體1〇〇的壽命，所述神經移植體還可進一步 包括一多聚賴氨酸層設置在所述神經網路20與所述蛋白 質層14之間。 請參閱圖6，所述神經網路20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 099143853 表單編號A0101 第21頁/共41頁 0992075908-0 [0049] 201224146 神經細胞22延伸出來的神經突起24的個數不限，只要能 夠使所述複數神經細胞22之时认生㈣接使所述複 數神經細胞22能夠相互溝通即可。譬如，其中—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。 闕本發明中的神經移植體100，具有修復生物體中神經系统 中的神經網路2Q設置在該培育層1G表面。輯述培育層 10中的疏水性基底丨丨具有較好的，具有*含金屬、彈性 佳、不㈣缺延祕良好㈣點。職培育層10中的 不米碳s膜結構1 2為基本由奈米碳管組成的自支推結構 丄具有不含金屬、彈性佳、枝顧、延展性良好及低 密度等優點。故，該培育層110可隨同該由複數神經突起 24連接的複數神經細胞22〜起植人到生物體中，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，植體1GG進行裁剪或拉伸。 闺以T將結合_纽具體實施财辆細㈣本發明的 神經移植體的製備方法及神經移植雜。 闕本發明提供—種神經移植體的製備方法其包括如下步 驟： [0053] S210 ’提供一矽膠基底。 刪麟碎膠基底的尺寸與厚度可根據實際需求而確定。譬 如，如果所需神經移植體的面積為3平方董米，則所述石夕 膠基底的面積可大於等於3平方复米。所述石夕膠基底不含 金屬，對生物體基本無毒性。 0992075908-0The immersion time of the s犋 structure is determined by the specific structure of the structure of the carbon nanotube membrane and the specific composition of the I-form solution, Form No. A0101, page 13 of 4192075908-0 201224146, as long as the nanometer can be made. Most of the carbon nanotubes in the carbon nanotube membrane structure are infiltrated with a protein solution. Typically, the carbon nanotube membrane structure has a soaking time of more than 2 hours. The environment in which the carbon nanotube membrane structure is immersed is not limited as long as the protein is not deteriorated. Usually, the soaking process can be carried out under normal temperature and normal pressure conditions. [0032] when the carbon nanotube membrane structure is immersed in the protein solution, the protein solution may infiltrate a portion of the carbon nanotubes in the carbon nanotube membrane structure, and may also infiltrate the nanometer tube All carbon nanotubes in the carbon tube membrane structure. Generally, when the thickness of the carbon nanotube film structure is thin, such as when the thickness of the carbon nanotube film structure is less than or equal to 10 μm, the protein solution may be infiltrated into the carbon nanotube film structure. All carbon nanotubes. 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. [0033] 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 make the protein in the protein solution not to be destroyed as described in Form 099143853 Form No. A0101 Page 14 of 41 0992075908-0 201224146, the temperature during sterilization should 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. [0034] In step S10, in order to increase the adhesion of the culture layer to the nerve cells and provide a growth environment more suitable for the nerve cells, after forming the protein layer, the step S10 may further include the following steps: S15, The protein layer forms a poly-D-lysine (PDL) layer away from the surface of the hydrophobic substrate. Specifically, the incubation layer may be immersed in a polylysine solution having a polylysine concentration of approximately 20 micrograms per milliliter. [0035] In step S20, the nerve cells include nerve cells of a mammal, and preferably, the nerve cells are hippocampal neurons. A 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 a surface of the cultivating layer away from the hydrophobic substrate, or the cultivating layer may be immersed in the In the nerve cell fluid containing nerve cells, the nerve cell solution may be covered with the growth layer. In order for the nerve cell fluid to cover the germ layer, the nerve cell fluid may be contained in a petri dish. The incubation layer can be suspended in the culture dish. It can also be disposed on a bottom surface of the culture dish as long as it can 099143853 Form No. A0101 Page 15 / Total 41 page 0992075908-0 201224146 The culture solution can be covered with the nerve cells. When the incubation layer is disposed on the bottom surface of the culture, 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. [0036] In step S30, the incubation environment of the nerve cells is not limited as long as the neurites can be grown. Generally, the nerve cells can grow in a normal temperature and a normal pressure environment. That is, the nerve cells are placed in an indoor environment, and the nerve cells can be grown, and proteins such as bovine serum in the culture layer can provide a growth factor to promote the growth of the nerve cells. Of course, the incubation environment may also be 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 simulated. [0037] The nerve cells are capable of growing a plurality of neurites 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 placed on the surface of the layer, the nerve cells of the nerve cells have a tendency to grow along adjacent nerve cells. So that adjacent nerve cells can connect and communicate. 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 the neurites connected between adjacent nerve cells are grown, the plurality of neural cells form the neural network by the neurites 099143853 Form No. A0101 Page 16 of 41 0992075908-0 201224146 , the plurality of nerve cells can communicate with each other. If the neurites of adjacent nerve cells meet, they will merge into the gods and dogs. For example, when the nerve cells are arranged in a line or array manner in the cultivating layer, and the gods t (four) in the longitudinal direction are closer to each other, and the nerve cells in the lateral direction are far apart, when: The nerve cells grown by the nuclear cells may be substantially along the longitudinal direction (for the purpose of enabling the nerve cells to be linear or (4) in the layer, the carbon nanotubes may be selected. ^ The carbon nanotubes extend substantially in the same direction. Through cultivation, most of the nerve cells of each other establish a connection through the thief, thereby describing the neural network and the formation of the neural network. a nerve graft. [0038] Ο The method for making a transplant is to provide a carbon nanotube membrane structure on a simple base surface, and then set the carbon nanotube membrane structure away from the surface of the hydrophobic substrate. The protein layer forms a cultivating layer, so that the plurality of phase cells can be planted in the protein layer φ in the cultivating layer, and the plurality of neurites are grown in the plurality of neurites. Can make Said # via the cell, that is, can not provide an environment for the growth of the cells, so that the nerve cells will only grow on the surface of the protein layer provided. [0039] The carbon nanotube membrane structure is A macroscopic membrane-like structure generally has an area of 15 mm x 15 mm or more. Specifically, the carbon nanotube membrane structure has a length of more than 3 GG meters and a width of more than 55 m. The substrate is also a 1 view structure, and its shape and surface area are adjusted according to the shape and area of the carbon nanotube film structure. 4 The carbon nanotube film 099143853 Form No. 1010101 Page 17 / Total 41 Page 0992075908-0 201224146 Structure The hydrophobic substrate has the advantages of good elasticity, good ductility and no metal, and can be directly implanted into the living body. Therefore, the nerve graft body mainly composed of the carbon nanotube film structure and the hydrophobic substrate is used. The damaged portion can be cut, stretched, and implanted according to the shape and size of the damaged portion of the damaged nervous system. The neural network has biological activity and signal transmission capability, thereby including the neural network. The transplant body also has biological activity and signal transmission ability. When the nerve graft is implanted into the damaged part of the organism, due to the neurons in the nerve implant and the ends or edges of the damaged part The distance between the neurons is short, so that the signal transmission ability can be established at both ends of the damaged portion by directly suturing the neurons in the nerve implant and the neurons at the edge of the damaged portion. The nerve repair of the damaged part, thereby saving the growth time of the neurite and reducing the repair time of the damaged nervous system. It can be understood that even when the nerve implant is implanted into the damaged part, no direct stitching is performed. Since the distance between the neurons at the edge of the damaged portion of the neuron in the nerve implant is smaller than the distance between the neurons at both ends of the damaged portion, the implant can also be reduced by implanting the nerve implant. The growth time of neurites, thereby reducing the repair time of the damaged nervous system. [0040] It should be noted that, in general, the carbon nanotubes in the structure of the carbon nanotube membrane refer to a carbon nanotube that has not been subjected to chemical or physical treatment, such as a nanometer that has 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. Nano carbon tube 099143853 Form No. A0101 Page 18 of 41 0992075908-0 201224146 Modification will not make any substantial contribution to the realization of this financial, because the protein shell layer covers the carbon nanotube The nerve cells are not in direct contact with the carbon nanotubes, and the surface structure of the carbon nanotubes is practically negligible. [0044] [0045] The nerve graft of the present invention may include a method for preparing the above-described nerve graft, which is cultured on the surface of the layer including the carbon nanotube membrane structure and the protein layer. A product obtained by neural networks formed by nerve cells and neurites. Referring to FIG. 5, the nerve graft 1 includes a layer 1 and a neural network 2分布 distributed on the surface of the layer. The incubation layer 10 includes a hydrophobic substrate, a carbon nanotube membrane structure 12, and a protein layer 14. The carbon nanotube film structure 2 is disposed on one or opposite surfaces of the hydrophobic substrate 1丨. The protein layer 14 is disposed on a surface of the carbon nanotube film structure 12 away from the hydrophobic substrate 11. In the present embodiment, the carbon nanotube structure 12 is disposed only on one of the facing surfaces of the hydrophobic substrate 11. The hydrophobic substrate 11 may be made of tantalum or coated with a gel. Tannin is a commonly used implant material for living organisms, which is non-toxic to organisms and has good flexibility. Therefore, the hydrophobic substrate 11 formed or coated with the gelatin 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 carbon nanotubes are connected to each other by van der Waals to form a self-supporting structure. The carbon nanotube membrane structure 12 comprises at least one carbon nanotube membrane, which can be a carbon nanotube as shown in FIG. 2 099143853 Form No. 1010101 Page 19/41 Page 0992075908-0 201224146 The flocculated membrane, the carbon nanotube rolled membrane of Figure 3, and the nanocarbon tubular membrane of Figure 4. In the present embodiment, the carbon nanotube film structure 12 includes a plurality of laminated films arranged in a plurality of layers, and adjacent drawn films are bonded to each other by van der Waals force. In the adjacent drawn film, the extending direction of the carbon nanotubes may have an angle of intersection, and preferably, the angle of intersection is 90 degrees. The thickness of the carbon nanotube film structure 12 can be set according to specific needs. 5微米小于小于微米微米。 Typically, the carbon nanotube film structure 12 thickness is greater than 0.3 microns less than 60 microns. 6微米。 The thickness of the carbon nanotube film structure 12 is substantially 0.6 microns. [0046] The protein layer 14 is composed of a soluble protein. The so-called soluble protein means that the protein has good hydrophilicity. The thickness of the protein layer 14 is not limited as long as a hydrophilic environment can be provided. 5微米至2微米。 Generally, the thickness of the protein layer 14 is 0.3 microns to 2 microns. 5微米。 The thickness of the protein layer is substantially 0.5 microns. Macroscopically, the protein layer 14 can optionally be disposed only on the surface of the carbon nanotube membrane structure 12 remote from the hydrophobic substrate 11. Microscopically, the protein in the protein layer 14 easily penetrates into the interior of the carbon nanotube membrane structure 12 and coats some or all of the carbon nanotubes in the carbon nanotube membrane structure 12, There is no distinct interface between the protein layer 14 and the carbon nanotube membrane 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 12 is less than or equal to 3 μm, the protein in the protein layer 14 easily penetrates into the The interior of the carbon nanotube membrane structure 12 substantially covers all of the carbon nanotubes in the carbon nanotube membrane structure 12. When the thickness of the carbon nanotube film structure 12 is thick, for example, the carbon nanotube film structure 12 099143853 Form No. A0101 Page 20 / Total 41 Page 0992075908-0 201224146 [0047] Ο ο [0048 When the thickness is greater than or equal to 3 microns, the protein in the protein layer 14 may also penetrate into the inner surface of the carbon nanotube membrane structure 12, but usually only the carbon nanotube membrane structure 12 is coated close to the Neural network 2 〇 carbon nanotubes. In this embodiment, the protein in the protein layer 14 substantially covers all of the carbon nanotubes in the carbon nanotube membrane structure 12. The neural network 20 is disposed on a surface of the protein layer 14 that is remote from the nanocarbon film structure 12. When the nerve graft 1 includes only one nanocarbon film structure 12 and the carbon nanotube film structure 12 is provided with a protein layer 14 away from the surface of the hydrophobic substrate 11, the nerve graft 10 0 includes only one surface of the neuron 20 disposed on the surface of the protein layer 丨4. When the nerve graft 100 includes two carbon nanotube membrane structures 丨2 respectively disposed on two surfaces of the hydrophobic substrate ,, and each of the carbon nanotube membrane structures 12 is disposed away from the surface of the hydrophobic substrate 11 When there is a protein, the nerve graft 100 may include two neural networks 2, respectively disposed on the surface s of the two protein layers 14, or may include only one neural network 20 disposed in one of the proteins. The surface of layer 14. In this embodiment, the nerve graft 100 includes only one carbon nanotube membrane structure 12, a protein layer 14 disposed on the surface of the carbon nanotube membrane structure 12, and a protein layer disposed on the protein layer. 14 surface neural network 20. It can be understood that, in order to improve the bacteriostasis of the biological graft 100 and increase the life span of the nerve graft, the nerve graft may further include a poly-lysine layer disposed on the neural network. 20 is between the protein layer 14. Referring to Figure 6, the neural network 20 includes a plurality of neural cells 22 and a plurality of neurites 24 extending from the plurality of neural cells 22. Each 099143853 Form No. A0101 Page 21 / Total 41 Page 0992075908-0 [0049] 201224146 The number of nerve protrusions 24 extended by the nerve cells 22 is not limited as long as the plurality of nerve cells 22 can be recognized (4) The plurality of nerve cells 22 can be communicated 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. The nerve graft 100 of the present invention has a neural network 2Q in the nervous system in the repairing organism disposed on the surface of the growth layer 1G. The hydrophobic substrate 辑 in the cultivating layer 10 is preferably a good one, and has a metal-containing, good elasticity, and no (four) defect. The non-rice carbon s film structure 12 in the occupational layer 10 is a self-supporting structure consisting essentially of carbon nanotubes. The crucible has the advantages of no metal, good elasticity, good branching, good ductility and low density. Therefore, the cultivating layer 110 can be implanted into the living body along with the plurality of nerve cells 22 connected by the plurality of neurites 24 for repairing the damaged nervous system in the living body, and can be based on the nervous system in the living body. The wound area is cut or stretched by the nerve, the implant 1GG.闺T will be combined with T-News implementation of the fourth embodiment of the invention, and a method for preparing a nerve graft of the present invention. The present invention provides a method of preparing a nerve graft comprising the steps of: [0053] S210' provides a silicone substrate. The size and thickness of the smashed base can be determined according to actual needs. For example, if the area of the desired nerve graft is 3 square meters, the area of the asphalt substrate may be greater than or equal to 3 square meters. The Shiqi gum substrate does not contain metal and is substantially non-toxic to the organism. 0992075908-0
099143853 表單編號 MUOi 第 22 1/共 41 I 201224146 剛S22G，將-奈米碳官膜結構鋪設在所述謂基底的—表 面。 _6]為使所述奈*碳管膜結構與所述鄉基底表面結合更緊 密，可對所述奈米碳管膜結構進行有機溶劑處理。具體 地’可在設置在所述梦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膜結構的比表面及增加 该奈米碳管膜結構與所述矽膠基底的附著力。所述奈米 0 碳管膜結構包括複數層奈米碳管拉膜，相鄰的奈米碳管 拉膜之間的奈米碳管的延伸方向具有一交叉角度。請參 閱圖7及圖8 ’優選地，所述交又角度大致尊於9〇度。 [0057] S230，將鋪設有奈米碳管膜結構的矽膠基尨浸泡在一蛋 白質溶液中。 [0058] 由於所述奈米碳管膜結構設置在所述矽膠基底表面，卷 所述奈米碳管膜結構連同矽膠基底一起浸,泡到所述純蛋 白質中時，所述奈米碳管膜結槔受液體表面張力影響而 〇 產生破損的概率賻笑為降低。所述蛋白質溶液純牛血清 溶液。請參見圖9 ’當所述奈米碳管膜結構從所逑蛋白質 溶液中浸泡1 · 5個小時左右，所述奈米碳管膜結構中的大 部分奈米碳管表面可浸潤有蛋白質溶液。 、大 剛S240，從所述蛋白質溶液中取出所述鋪設有奈米碳管棋 結構的矽膠基底，在120攝氏度下進行高溫滅菌處理’， 成一培育層。 形 [0060]將所述鋪設有奈米碳管膜結構的矽膠基底從所述蛋白 099143853 表單煸號A0101 第23頁/共41頁 201224146 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，滅菌處理後後， 所述蛋白質溶液中的蛋白質基本固化，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。 [0061] [0062] [0063] [0064] [0065] [0066] 099143853 S250 ’將所述培育層浸泡在一聚賴氨酸溶液中。 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。 S26〇，在所述浸泡後的多育層滴加一神經細胞液直到該 神經細胞液復蓋該培育層。 由於所述培育層中具有矽膠基底，該培育層可直接設置 在一容器如培養皿中，通過所述矽膠基底係所述奈米碳 管膜結構不與所述容器的内表面接觸。 S270，培育所述附著在所述培育層的複數神經細胞使 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，從而在所述培育層形赛一神經網路。 所述神經細胞的培育環境為普通的室内環境，培育時間 可根據實際需求而定。故，在步驟326〇的環境下，請參 閱圖10，保持各種條件不變，在室内環境培養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 經細胞生長的生長因數。所述多個神經細胞上的多個神 經突起相互連接後，形成所述神經網路及移植體，請參 閱圖11及圖12，為所述神經移植體未經染色的掃描電鏡 表單編號A0101 第24頁/共41頁 201224146 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，如圖1 1所示，部分神經 細％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。 [0067] Ο [0068] Ο [0069] 需要指出的係，由於所述培育層中的矽膠基底將所述奈 米嘰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碳 督螟結構上蛋白質層表面且在該表面生長，而不會在所 迷矽膠基底表面或容器的内表面上生長》 、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a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Q 【圖式簡單說明】 圖1為本發明實施例所提供的—神經移植體的製備方法的 流程示意圖。 [0070] [0071] [0072] [0073] 圖2為一奈米碳管絮化膜的掃描電鏡照片。 圖3為一奈米碳管碾壓膜的掃描電鏡照片。 圖4為一奈米碳管拉骐的掃描電鏡照片。 圖5為本發明實施例所提供的神經移植體的侧視示意圖 099143853 0992075908-0 表單編號A0101 第25頁/共41頁 201224146 [0074] 圖6為本發明實施例所提供的神經移植體的俯視示意圖。 [0075] 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。 [0076] 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。 [0077] 圖9為本發明實施例所提供的培育層的透射電鏡照片。 [0078] 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。 [0079] 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。 [0080] 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】 [0081] 神經移植體：100 [0082] 培育層：10 [0083] 疏水性基底：11 [0084] 奈米碳管膜結構：1 2 [0085] 蛋白質層：1 4 [0086] 神經網路：20 [0087] 神經細胞：22 [0088] 神經突起：24 099143853 表單編號A0101 第26頁/共41頁 0992075908-0099143853 Form No. MUOi No. 22 1 / 41 I 201224146 Just S22G, the -Nano carbon film structure is laid on the surface of the said substrate. _6] The carbon nanotube film structure may be subjected to an organic solvent treatment in order to make the structure of the carbon nanotube film more tightly bonded to the surface of the substrate. Specifically, the carbon nanotube film can be reduced by covering or depositing a solvent which is easily volatilized, such as an organic solvent, on the surface of the monical base substrate, and then volatilizing the solvent. The specific surface of the structure and the adhesion of the carbon nanotube film structure to the silicone substrate. The nano 0 carbon tube membrane structure comprises a plurality of layers of carbon nanotube membranes, and the direction of extension of the carbon nanotubes between adjacent carbon nanotube membranes has an intersection angle. Please refer to Figures 7 and 8'. Preferably, the angle of intersection is approximately 9 degrees. [0057] S230, soaking the silicone-based crucible having the structure of the carbon nanotube film in a protein solution. [0058] Since the carbon nanotube film structure is disposed on the surface of the silicone substrate, the carbon nanotube film structure is immersed together with the silicone substrate, and the carbon nanotube is bubbled into the pure protein. The film crucible is affected by the surface tension of the liquid and the probability of breakage is reduced. The protein solution is a pure bovine serum solution. Please refer to FIG. 9 'When the carbon nanotube membrane structure is immersed from the sputum protein solution for about 1.5 hours, most of the surface of the carbon nanotube membrane can be infiltrated with a protein solution. . , Dagang S240, the silicone substrate having the nanocarbon tube structure is taken out from the protein solution, and subjected to high temperature sterilization treatment at 120 degrees Celsius to form a cultivation layer. Forming [0060] the silicone substrate having the carbon nanotube membrane structure is taken out from the solution of the protein 099143853 Form No. A0101, page 23 of 41201224146, and can be heat-sterilized in a dry box. . The sterilization temperature of the drying oven is approximately 120 degrees. After the sterilization treatment, the protein in the protein solution is substantially solidified, and a protein layer is formed on the surface of the carbon nanotube membrane structure to form the incubation layer. [0063] [0066] [0066] 099143853 S250 'The incubation layer was immersed in a polylysine solution. 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. S26〇, a nerve cell fluid is added dropwise to the soaked multi-culture layer until the nerve cell fluid covers the culture layer. Since the cultivating layer has a silicone substrate, the cultivating layer can be directly disposed in a container such as a petri dish through which the carbon nanotube film structure is not in contact with the inner surface of the container. S270, cultivating the plurality of nerve cells attached to the cultivating layer to grow the plurality of nerve cells to grow a plurality of neurites connected between the plurality of nerve cells, thereby forming a neural network in the cultivating layer. The cultivation environment of the nerve cells is an ordinary indoor environment, and the cultivation time can be determined according to actual needs. Therefore, in the environment of step 326, please refer to Fig. 10, and keep the various conditions unchanged. After culturing for about 15 days in an indoor environment, the nerve cells can be differentiated into a plurality of 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 neural cells. After the plurality of nerve cells on the plurality of nerve cells are connected to each other, the neural network and the implant are formed. Referring to FIG. 11 and FIG. 12, the nerve graft is not stained by SEM form No. A0101. 24 pages/total 41 pages 201224146 Photos and TEM images after staining. It can be clearly seen from the above-described projection electron microscope photograph that a plurality of nerve cells in the nerve graft are connected together by neurites. At the same time, as shown in Fig. 11, part of the nerve fineness extends out of multiple neurites but does not connect with other nerve cells through the multiple neurites. But this does not affect the nerve graft as a whole. Biologically active. [0067] [0069] It is noted that the tantalum substrate itself has water repellency since the silicone substrate in the incubation layer separates the container of the nanotube membrane structure. Therefore, the nerve cells will only adsorb on the surface of the protein layer disposed on the nano-carbon structure and grow on the surface without growing on the surface of the gelatin substrate or the inner surface of the container. As described, the present invention has indeed met the requirements of the invention patent, and the patent application is filed according to law. However, the above-mentioned preferred embodiments of the present invention are not intended to limit the scope of the present invention. Any equivalent modifications or variations made by those skilled in the art in the light of the present invention should be covered. In the following patent application, the following is a schematic diagram of a method for preparing a nerve graft. [0073] FIG. 2 is a scanning electron micrograph of a carbon nanotube flocculation membrane. Figure 3 is a scanning electron micrograph of a carbon nanotube rolled film. Figure 4 is a scanning electron micrograph of a carbon nanotube pull. FIG. 5 is a side view of a nerve graft according to an embodiment of the present invention. 099143853 0992075908-0 Form No. A0101 Page 25 of 41 201224146 [0074] FIG. 6 is a plan view of a nerve graft provided by an embodiment of the present invention. schematic diagram. 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. 12 is a scanning electron micrograph of a nerve graft after staining according to an embodiment of the present invention. [Key element symbol description] [0081] Nerve graft: 100 [0082] Cultivating layer: 10 [0083] Hydrophobic substrate: 11 [0084] Nano carbon tube membrane structure: 1 2 [0085] Protein layer: 1 4 [ 0086] Neural Network: 20 [0087] Nerve Cells: 22 [0088] Nerve Protrusion: 24 099143853 Form No. A0101 Page 26 of 41 0992075908-0