Gorgieva Selestina, Štrancar Janez, Kokol Vanja
Faculty of Mechanical Engineering, Institute for Engineering Materials and Design, University of Maribor, Maribor, Slovenia.
J Biomed Mater Res A. 2014 Nov;102(11):3986-97. doi: 10.1002/jbm.a.35076. Epub 2014 Jan 6.
This article present new insights in complex relation between surface- and interface-related physicochemical properties and microstructuring of three-dimensional (3D) gelatin scaffolds, being fabricated by simultaneous temperature-controlled freeze-thawing cycle and in situ cross-linking using variable conditions (pH) and molarity of carbodiimide reagents, on the seeding and growth of fibroblast cells with subsequent tracking of their spreading and morphology. Rarely populated cells with rounded morphology and small elongations are observed on negative charge-rich scaffold surface with a lower cross-linking degree (CD), and consequently higher molecular mobility and availability of cell-recognition sequences, in comparison with the prominently elongated and densely populated cells on a positively charged scaffold's surface with higher CD and low mobility. Surface microstructure effect was demonstrated by cell vacuolization and their pure intercommunication being present on scaffold's bottom side with smaller pores (25 ± 19 µm) and pore wall thickness (9 ± 5 µm), over the air-exposed side with twice bigger pores (56 ± 38 µm) and pore wall thicknesses (12 ± 6 µm). Strong correlations of CD (r(2) = 0.96) and local molecular mobility (r(2) = -0.44) with pH and reagents molarity, as well as microstructure features being related to temperature gradient, imply on possibility to modulate scaffold's properties in a direction to guide cell viability and most likely its genotype development.
本文介绍了三维(3D)明胶支架的表面和界面相关物理化学性质与微观结构之间复杂关系的新见解。该支架通过同时进行温度控制的冻融循环以及使用可变条件(pH)和碳二亚胺试剂的摩尔浓度进行原位交联来制备,研究了其对成纤维细胞接种和生长的影响,并随后跟踪了细胞的铺展和形态。与具有较高交联度(CD)和低流动性的带正电支架表面上显著伸长且密集分布的细胞相比,在交联度较低、因而分子流动性较高且细胞识别序列可用性较高的富含负电荷的支架表面上,观察到形态呈圆形且伸长较小的细胞分布较少。通过细胞空泡化以及在孔径较小(25±19μm)和孔壁厚度较薄(9±5μm)的支架底部一侧存在的纯细胞间连通,与暴露于空气一侧具有两倍大的孔径(56±38μm)和孔壁厚度(12±6μm)相比,证明了表面微观结构效应。CD(r² = 0.96)和局部分子流动性(r² = -0.44)与pH和试剂摩尔浓度之间的强相关性,以及与温度梯度相关的微观结构特征,意味着有可能在引导细胞活力及其最可能的基因型发展的方向上调节支架的性质。