本文選題：3D打印技術　切入點：氣管補片　出處：《揚州大學》2017年碩士論文　論文類型：學位論文

【摘要】：氣管切除并端端吻合是當前氣管重建的金標準,但僅限于病變氣管段不超過成人總氣管長度1/2或小兒1/3。當病變氣管超過最大限度時,進行氣管重建有一定難度,因為氣管不僅僅是一個簡單的圓柱狀通氣管道,而是由復雜的多層結構組成。氣管是由15-20個C形軟骨構成,在氣管內(nèi)表面覆有纖毛上皮,外表面含有平滑肌、血管等結締組織。氣管軟骨維持著氣管圓柱狀形態(tài),防止氣管塌陷;而氣管內(nèi)表面呼吸上皮中的纖毛對氣管的清潔有著重要作用;氣管軟骨周圍的結締組織則保證了氣管的曲、伸以及收縮、擴張等機械運動。因此,還沒有辦法完全重建這樣復雜的多層結構以及完全模擬其功能。近年來,3D打印技術的發(fā)展為氣管重建提供了新的思路。3D打印技術依靠計算機輔助成像,以廣泛使用的生物材料為打印介質(zhì),能夠快速、精確的復制和重建缺損組織或器官的復雜結構,因此在組織工程領域的應用獲得廣泛關注。骨髓間充質(zhì)干細胞易分離培養(yǎng),且具有多向分化潛能,是氣管組織工程首選的種子細胞。本研究旨在以聚己內(nèi)酯為材料,利用3D打印技術打印出氣管補片,通過生物力學測試評估其生物力學性能并與骨髓間充質(zhì)干細胞共培養(yǎng)評估其細胞相容性,從而尋求合適的組織工程氣管支架材料。第一部分3D打印氣管補片的制備與生物力學性能檢測目的:利用3D打印技術將聚己內(nèi)酯打印成氣管補片并研究其生物力學性能。方法:1.3D打印氣管補片的制備;2.掃描電子顯微鏡觀察3D打印氣管補片超微結構;3.3D打印氣管補片的生物力學性能測試。結果:1.掃描電子顯微鏡圖(SEM)觀察到3D打印氣管補片擁有適宜的孔徑大小,孔徑為 300-500μm;2.生物力學性能測試結果證實3D打印氣管補片的最大應力及彈性模量明顯優(yōu)于離體新鮮氣管,顯示其具有良好的生物力學性能。結論:1.利用3D打印技術將聚己內(nèi)酯制備成3D打印氣管補片;2.3D打印氣管補片具備合理的三維外形、適宜的孔徑大小;3.3D打印氣管補片具備良好的生物力學性能。第二部分 骨髓間充質(zhì)干細胞體外培養(yǎng)與3D打印氣管補片細胞相容性檢測目的:1.骨髓間充質(zhì)干細胞的體外培養(yǎng);2.檢測3D打印氣管補片與骨髓間充質(zhì)干細胞共培養(yǎng)的細胞相容性。方法:1.兔骨髓間充質(zhì)干細胞的獲取;2.兔骨髓間充質(zhì)干細胞的培養(yǎng);3.兔骨髓間充質(zhì)干細胞的鑒定;4.3D打印氣管補片與骨髓間充質(zhì)干細胞共培養(yǎng)的細胞相容性檢測。結果:1.通過全骨髓培養(yǎng)及貼壁純化法獲取的骨髓間充質(zhì)干細胞,培養(yǎng)至第3代細胞成簇貼壁生長,呈梭形、多角形,具有多向分化潛能,可以分化為軟骨細胞和脂肪細胞;2.與骨髓間充質(zhì)干細胞共培養(yǎng)后進行細胞相容性檢測結果顯示3D打印氣管補片具有良好的細胞相容性。結論:3D打印氣管補片具備良好的細胞相容性,是一種具有開發(fā)潛力的生物材料,可以用于組織工程氣管的體外構建。
[Abstract]:Trachea resection with end-to-end anastomosis is the golden standard for trachea reconstruction, but it is only limited to the total trachea length of 1/2 in adults or 1 / 3 in children. Because the trachea is not just a simple cylindrical tube, but it's made up of a complex, multilayered structure. The trachea is composed of 15-20 C-shaped cartilage, covered with cilia epithelium on the surface of the trachea, and smooth muscle on the outer surface. Connective tissue such as blood vessels. The trachea cartilage maintains the cylindrical shape of the trachea and prevents the collapse of the trachea; the cilia in the respiratory epithelium on the surface of the trachea play an important role in the cleaning of the trachea; the connective tissue around the cartilage of the trachea ensures the curvature of the trachea. Extension and mechanical movements such as contraction and expansion. There is no way to completely reconstruct such a complex multilayer structure and to completely simulate its functions. In recent years, the development of 3D printing technology has provided a new way of thinking for trachea reconstruction. 3D printing technology relies on computer-aided imaging. Using widely used biomaterials as printing media, the complex structures of defective tissues or organs can be reproduced and reconstructed quickly and accurately, so their applications in the field of tissue engineering have attracted wide attention. Bone marrow mesenchymal stem cells are easily isolated and cultured. The aim of this study was to use polycaprolactone as the material, and to print the trachea patch with 3D printing technology. Its biomechanical properties were evaluated by biomechanical tests and its cytocompatibility was evaluated by co-culture with bone marrow mesenchymal stem cells. In order to find suitable tissue engineering tracheal scaffold materials. Part I preparation of 3D printed trachea patch and biomechanical properties test objective: to print polycaprolactone into trachea patch by 3D printing technology and study its biological force. Methods: 1. Preparation of 3D printed trachea patch. Observation of ultrastructure of 3D printed trachea patch by scanning electron microscope. 3. Biomechanical properties of 3D printed trachea patch. Results: 1. Scanning electron microscopy (SEM) observation of 3D trachea patch. The printed trachea patch has an appropriate aperture, The maximum stress and elastic modulus of 3D printed trachea patch were obviously superior to those of fresh trachea in vitro, the results of biomechanical test showed that the maximum stress and elastic modulus of 3D printed tracheal patch were obviously superior to those of fresh trachea in vitro. Conclusion: 1. Using 3D printing technology to prepare 3D printed trachea patch 2.3D printed trachea patch has a reasonable 3D shape. Suitable aperture size 3. 3D printed trachea patch has good biomechanical properties. Part 2: bone marrow mesenchymal stem cells cultured in vitro and 3D printed trachea patch cytocompatibility test objective: 1. Bone marrow mesenchymal stem cells. The cytocompatibility of 3D printed tracheobronchial patch and bone marrow mesenchymal stem cells was examined in vitro. Methods: 1. Acquisition of rabbit bone marrow mesenchymal stem cells 2. Culture of rabbit bone marrow mesenchymal stem cells 3. Fine mesenchymal stem cells of rabbit bone marrow. Cytocompatibility of 3D printed tracheal patch and bone marrow mesenchymal stem cells. Results: 1. Bone marrow mesenchymal stem cells obtained by whole bone marrow culture and adherent purification. In the third generation, the cells grew in clusters, fusiform and polygonal, and had the potential of multidirectional differentiation. It can differentiate into chondrocytes and adipocytes. After co-culture with bone marrow mesenchymal stem cells, the results of cytocompatibility test show that 3D printed trachea patch has good cytocompatibility. Good cell compatibility, It is a potential biomaterial for in vitro construction of tissue engineering trachea.
【學位授予單位】：揚州大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TP391.73;R318.08

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