Baker Simon C, Atkin Neil, Gunning Paul A, Granville Nick, Wilson Karen, Wilson Darren, Southgate Jennifer
Jack Birch Unit of Molecular Carcinogenesis, Department of Biology, University of York, Heslington, York, YO10 5YW, UK.
Biomaterials. 2006 Jun;27(16):3136-46. doi: 10.1016/j.biomaterials.2006.01.026. Epub 2006 Feb 10.
The purpose of this study was to produce a well-characterised electrospun polystyrene scaffold which could be used routinely for three-dimensional (3D) cell culture experimentation. A linear relationship (p<0.01) between three principal process variables (applied voltage, working distance and polymer concentration) and fibre diameter was reliably established enabling a mathematical model to be developed to standardise the electrospinning process. Surface chemistry and bulk architecture were manipulated to increase wetting and handling characteristics, respectively. X-ray photoelectron spectroscopy (XPS) confirmed the presence of oxygen-containing groups after argon plasma treatment, resulting in a similar surface chemistry to treated tissue culture plastic. The bulk architecture of the scaffolds was characterised by scanning electron microscopy (SEM) to assess the alignment of both random and aligned electrospun fibres, which were calculated to be 0.15 and 0.66, respectively. This compared to 0.51 for collagen fibres associated with native tissue. Tensile strength and strain of approximately of 0.15 MPa and 2.5%, respectively, allowed the scaffolds to be routinely handled for tissue culture purposes. The efficiency of attachment of smooth muscle cells to electrospun scaffolds was assessed using a modified 3-[4,5-dimethyl(thiazol-2yl)-3,5-diphery] tetrazolium bromide assay and cell morphology was assessed by phalloidin-FITC staining of F-actin. Argon plasma treatment of electrospun polystyrene scaffold resulted in significantly increased cell attachment (p<0.05). The alignment factors of the actin filaments were 0.19 and 0.74 for the random and aligned scaffold respectively, compared to 0.51 for the native tissue. The data suggests that electrospinning of polystyrene generates 3D scaffolds which complement polystyrene used in 2D cell culture systems.
本研究的目的是制备一种特性良好的电纺聚苯乙烯支架，该支架可常规用于三维（3D）细胞培养实验。可靠地建立了三个主要工艺变量（施加电压、工作距离和聚合物浓度）与纤维直径之间的线性关系（p<0.01），从而能够开发一个数学模型来规范电纺工艺。分别对表面化学性质和整体结构进行了处理，以提高润湿性和可操作性。X射线光电子能谱（XPS）证实了氩等离子体处理后含氧基团的存在，从而产生了与经处理的组织培养塑料相似的表面化学性质。通过扫描电子显微镜（SEM）对支架的整体结构进行表征，以评估随机排列和定向排列的电纺纤维的排列情况，经计算其排列系数分别为0.15和0.66。相比之下，与天然组织相关的胶原纤维的排列系数为0.51。支架的拉伸强度和应变分别约为0.15MPa和2.5%，这使得该支架能够常规用于组织培养目的。使用改良的3-[4,5-二甲基（噻唑-2-基）-3,5-二苯基]溴化四氮唑法评估平滑肌细胞在电纺支架上的附着效率，并通过用F-肌动蛋白的鬼笔环肽-FITC染色来评估细胞形态。氩等离子体处理电纺聚苯乙烯支架导致细胞附着显著增加（p<0.05）。随机排列和定向排列的支架中肌动蛋白丝的排列系数分别为0.19和0.74，相比之下天然组织的排列系数为0.51。数据表明，聚苯乙烯的电纺产生了3D支架，可补充二维细胞培养系统中使用的聚苯乙烯。
