Tresoldi Claudia, Stefani Ilaria, Ferracci Gaia, Bertoldi Serena, Pellegata Alessandro F, Farè Silvia, Mantero Sara
Department of Chemistry, Materials, and Chemical Engineering "Giulio Natta", Politecnico di Milano, Milan - Italy.
INSTM Local Unit, Politecnico di Milano, Milan - Italy.
J Appl Biomater Funct Mater. 2017 Apr 26;15(2):e122-e132. doi: 10.5301/jabfm.5000334.
In vitro dynamic culture conditions play a pivotal role in developing engineered tissue grafts, where the supply of oxygen and nutrients, and waste removal must be permitted within construct thickness. For tubular scaffolds, mass transfer is enhanced by introducing a convective flow through rotating bioreactors with positive effects on cell proliferation, scaffold colonization and extracellular matrix deposition. We characterized a novel polyurethane-based tubular scaffold and investigated the impact of 3 different culture configurations over cell behavior: dynamic (i) single-phase (medium) rotation and (ii) double-phase exposure (medium-air) rotation; static (iii) single-phase static culture as control.
A new mixture of polyol was tested to create polyurethane foams (PUFs) as 3D scaffold for tissue engineering. The structure obtained was morphologically and mechanically analyzed tested. Murine fibroblasts were externally seeded on the novel porous PUF scaffold, and cultured under different dynamic conditions. Viability assay, DNA quantification, SEM and histological analyses were performed at different time points.
The PUF scaffold presented interesting mechanical properties and morphology adequate to promote cell adhesion, highlighting its potential for tissue engineering purposes. Results showed that constructs under dynamic conditions contain enhanced viability and cell number, exponentially increased for double-phase rotation; under this last configuration, cells uniformly covered both the external surface and the lumen.
The developed 3D structure combined with the alternated exposure to air and medium provided the optimal in vitro biochemical conditioning with adequate nutrient supply for cells. The results highlight a valuable combination of material and dynamic culture for tissue engineering applications.
体外动态培养条件在工程化组织移植物的开发中起着关键作用，在构建物厚度范围内必须允许氧气和营养物质的供应以及废物的清除。对于管状支架，通过旋转生物反应器引入对流可以增强传质，对细胞增殖、支架定植和细胞外基质沉积具有积极影响。我们对一种新型聚氨酯基管状支架进行了表征，并研究了3种不同培养配置对细胞行为的影响：动态（i）单相（培养基）旋转和（ii）双相暴露（培养基-空气）旋转；静态（iii）单相静态培养作为对照。
测试了一种新的多元醇混合物，以制备聚氨酯泡沫（PUF）作为组织工程的3D支架。对获得的结构进行形态学和力学分析测试。将小鼠成纤维细胞接种在新型多孔PUF支架外部，并在不同动态条件下培养。在不同时间点进行活力测定、DNA定量、扫描电子显微镜（SEM）和组织学分析。
PUF支架具有有趣的力学性能和足以促进细胞粘附的形态，突出了其在组织工程中的应用潜力。结果表明，动态条件下的构建物具有更高的活力和细胞数量，双相旋转时呈指数增加；在最后这种配置下，细胞均匀地覆盖了外表面和管腔。
所开发的3D结构与空气和培养基的交替暴露相结合，为细胞提供了具有充足营养供应的最佳体外生化条件。结果突出了材料与动态培养在组织工程应用中的宝贵结合。
