Fabrication of Nanofiber Reinforced Protein Structures For Tissue Engineering.
Fabrication of Nanofiber Reinforced Protein Structures For Tissue Engineering.
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DOI：
10.1016/j.msec.2009.07.008
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发表时间：
2009-10-15
影响因子：
7.9
通讯作者：
Wen, Xuejun
中科院分区：
文献类型：
--
作者：
Beachley, Vince;Wen, Xuejun
细胞外基质和可降解纳米纤维是组织工程领域中两种非常有前途的材料，然而这两种结构作为组织工程支架都面临着局限性。细胞外基质，如胶原蛋白、明胶、层粘连蛋白，具有良好的生物相容性，并允许细胞生长和存活，但结构上的弱点使它们难以处理，极大地限制了它们的应用。可降解的纳米纤维支持细胞附着，并可以提供结构支撑和方向引导，但单个可降解的纳米纤维是脆弱的，并具有形成致密纤维束的趋势，这限制了细胞渗透到纳米纤维之间的空间中，特别是在对齐的纳米纤维的情况下。为了克服这些困难，将可降解的松散纳米纤维嵌入蛋白质基质中，试图制造具有改进性能的杂化支架，例如改进的强度、引导性、纳米纤维之间的间距等。聚己内酯（PCL）被用作可降解纳米纤维的模型材料。明胶被用作基质结构形成的模型蛋白质。通过用明胶水溶液润湿松散排列的无方向性的纳米纤维阵列或松散排列的双向纳米纤维网格来制造薄的混合膜（平均厚度= 2.78 μ m），如果细胞与蛋白质溶液预混合或在膜的表面上，这也允许活细胞加载到纳米纤维-蛋白质复合物中。由于薄膜的脆弱性，单独的明胶膜不具有抗渗增强，因此不能处理。纤维密度低至3%v/v的明胶膜在结构上足够坚固，可用于处理和加工成复杂形状。力学测试证实，纳米纤维的加入增强了明胶膜在干燥和水合状态下的强度。体外试验证实，纤维增强膜具有生物相容性，并为细胞提供定向引导。结果表明，明胶/聚己内酯复合材料作为组织支架材料的承诺。
Extracellular matrices and degradable nanofibers are two very promising materials in the field of tissue engineering; however both of these structures face limitations as tissue engineering scaffolds. Extracellular matrices, such as collagen, gelatin, laminin, have excellent biocompatibility and allow cell in growth and survival, but structural weakness makes them difficult to handle and greatly limits their uses. Degradable nanofibers support cell attachment and can provide structural support and directional guidance, but individual degradable nanofibers are fragile and have a tendency to form dense fiber bundles which limited the cell penetration into the spaces between the nanofibers, especially in the case of aligned nanofibers. To overcome these difficulties, degradable loose nanofibers were embedded in protein matrix in an attempt to fabricate a hybrid scaffold with improved properties, such as improved strength, guidance, spacing among nanofibers, etc.. Polycaprolactone (PCL) was used as a model material for degradable nanofibers. Gelatin was employed as a model protein for matrix structure formation. Thin hybrid films (average thickness = 2.78 um) were fabricated by wetting the loose aligned undirectional nanofiber arrays or loose aligned bidirectional nanofiber grids with a gelatin aqueous solution, which also allows for live cell loading into the nanofiber-protein composite if cell are premixed with protein solution or on the surface of the films. Gelatin film alone without nanofiber reinforcement cannot be handled due to the weakness of the thin membrane. Gelatin films with a fiber density as low as 3% v/v were structurally robust enough for handling, and manipulation into complex shapes. Mechanical testing confirmed that the addition of nanofibers enhanced the strength of gelatin films, in both dry and hydrated state. In vitro testing confirmed that nanofiber reinforced films were biocompatible and provided cells with directional guidance. Results demonstrate the promise of gelatin/PCL nanofiber composites as a tissue scaffolding material.
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