Novel system for engineering bioartificial tendons and application of mechanical load
Novel system for engineering bioartificial tendons and application of mechanical load
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DOI：
10.1089/107632703322495619
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发表时间：
2003-10-01
影响因子：
--
通讯作者：
Banes, AJ
中科院分区：
文献类型：
--
作者：
Garvin, J;Qi, B;Banes, AJ
在三维胶原凝胶中培养的细胞表达更天然的状态表型，因为它们形成可以机械加载的合胞体网络。此外，细胞通过消除水以及通过重组和排列胶原纤维来重塑其基质。最后，使细胞在天然基质中经受机械加载的能力是期望的，因为组织以及基质中的细胞携带菌株并改变其表达谱，这与固定化、中等活性或重复加载一致。这是第一次报告的模型生物反应器系统，以制造和培养肌腱细胞填充，线性，拴系矩阵结构，可以通过计算机驱动，压力控制系统机械加载。作为肌腱结构的生物人工组织（BAT）在新型橡胶底组织训练培养板中成型，该培养板在每个培养孔的东极和西极带有非织造尼龙网锚。通过在六孔培养板的每个孔下方放置Arctangle加载柱（Arctangle是具有弯曲短端的矩形）并使用真空将柔性膜向下移位，从而在BAT上产生单轴应变来实现机械加载。填充有禽屈肌腱细胞的BAT表达胶原蛋白基因I、III和XII以及聚集蛋白聚糖、纤连蛋白、脯氨酰羟化酶和腱生蛋白，这与在胶原蛋白结合的二维表面上或在天然的、完整的禽屈肌腱中生长的细胞的表达水平一致。同样，在BAT的细胞建立了一个形态的线性排列的细胞与主应变方向一致，在整个肌腱的筋膜。最后，在培养的第一周，机械负载的BAT的极限拉伸强度几乎是未负载BAT的3倍。总之，这些结果表明，肌腱细胞制造的机械加载，线性胶原蛋白凝胶构建体假定的表型，是类似于天然肌腱的外观和表达方面，比未行使的同行，但远弱于天然成人肌腱。该技术代表了一种在机械活性的三维培养环境中培养细胞的新方法，该方法可以容易地用于制造用于药物测试或组织工程的组织模拟物。
Cells cultured in three-dimensional collagen gels express a more native state phenotype because they form a syncytial network that can be mechanically loaded. Moreover, cells remodel their matrix by eliminating water, and by reorganizing and aligning the collagen fibrils. Last, the ability to subject cells to mechanical loading in a native matrix is desirable because cells, in tissues as well as the matrix, bear strains and alter their expression profile consistent with either immobilization, moderate activity, or repetitive loading. This is the first report of a model bioreactor system to fabricate and culture tendon cell-populated, linear, tethered matrix constructs that can be mechanically loaded by a computer-driven, pressure-controlled system. Bioartificial tissues (BATs) as tendon constructs were molded in a novel, rubber bottom Tissue Train culture plate bearing nonwoven nylon mesh anchors at the east and west poles of each culture well. Mechanical loading was achieved by placing an Arctangle loading post (an Arctangle is a rectangle with curved short ends) beneath each well of the six-well culture plate and using vacuum to displace the flexible membrane downward, resulting in uniaxial strain on the BAT. BATs populated with avian flexor tendon cells expressed collagen genes I, III, and XII as well as aggrecan, fibronectin, prolyl hydroxylase, and tenascin, consistent with expression levels of cells grown on collagen-bonded two-dimensional surfaces or in native, whole, avian flexor tendon. Likewise, cells in BATs established a morphology of linearly arranged cells aligned with the principal strain direction as in fasicles of whole tendons. Last, BATs that were mechanically loaded had an ultimate tensile strength that was nearly 3-fold greater than that of nonloaded BATs in the first week of culture. Taken together, these results indicate that tendon cells fabricated in a mechanically loaded, linear collagen gel construct assume a phenotype that is similar to that of a native tendon in terms of appearance and expression and are stronger than nonexercised counterparts yet far weaker than native adult tendons. This technique represents a novel approach to culturing cells in a mechanically active, three-dimensional culture environment that can be readily used for the fabrication of tissue simulates for drug testing or tissue engineering.