Williamson Matthew R, Woollard Kevin J, Griffiths Helen R, Coombes Allan G A
Aston Pharmacy School, Aston University, Aston Triangle, Birmingham, United Kingdom.
Tissue Eng. 2006 Jan;12(1):45-51. doi: 10.1089/ten.2006.12.45.
Poly(epsilon-caprolactone) (PCL) fibers produced by wet spinning from solutions in acetone under lowshear (gravity-flow) conditions resulted in fiber strength of 8 MPa and stiffness of 0.08 Gpa. Cold drawing to an extension of 500% resulted in an increase in fiber strength to 43 MPa and stiffness to 0.3 GPa. The growth rate of human umbilical vein endothelial cells (HUVECs) (seeded at a density of 5 x 10(4) cells/mL) on as-spun fibers was consistently lower than that measured on tissue culture plastic (TCP) beyond day 2. Cell proliferation was similar on gelatin-coated fibers and TCP over 7 days and higher by a factor of 1.9 on 500% cold-drawn PCL fibers relative to TCP up to 4 days. Cell growth on PCL fibers exceeded that on Dacron monofilament by at least a factor of 3.7 at 9 days. Scanning electron microscopy revealed formation of a cell layer on samples of cold-drawn and gelatin-coated fibers after 24 hours in culture. Similar levels of ICAM-1 expression by HUVECs attached to PCL fibers and TCP were measured using RT-PCR and flow cytometry, indicative of low levels of immune activation. Retention of a specific function of HUVECs attached to PCL fibers was demonstrated by measuring their immune response to lipopolysaccharide. Levels of ICAM-1 expression increased by approximately 11% in cells attached to PCL fibers and TCP. The high fiber compliance, favorable endothelial cell proliferation rates, and retention of an important immune response of attached HUVECS support the use of gravity spun PCL fibers for three-dimensional scaffold production in vascular tissue engineering.
通过在低剪切(重力流)条件下从丙酮溶液中湿法纺丝制备的聚(ε-己内酯)(PCL)纤维,其纤维强度为8 MPa,刚度为0.08 GPa。冷拉伸至500%的伸长率后,纤维强度增加到43 MPa,刚度增加到0.3 GPa。在第2天之后,接种密度为5×10⁴个细胞/mL的人脐静脉内皮细胞(HUVECs)在初纺纤维上的生长速率始终低于在组织培养塑料(TCP)上测得的生长速率。在7天内,明胶包被的纤维和TCP上的细胞增殖情况相似,在长达4天的时间里,相对于TCP,500%冷拉伸PCL纤维上的细胞增殖高出1.9倍。在第9天,PCL纤维上的细胞生长比涤纶单丝上的细胞生长至少高出3.7倍。扫描电子显微镜显示,培养24小时后,冷拉伸和明胶包被的纤维样品上形成了细胞层。使用RT-PCR和流式细胞术测量附着在PCL纤维和TCP上的HUVECs的ICAM-1表达水平相似,表明免疫激活水平较低。通过测量附着在PCL纤维上的HUVECs对脂多糖的免疫反应,证明了其特定功能的保留。附着在PCL纤维和TCP上的细胞中ICAM-1表达水平增加了约11%。高纤维顺应性、有利的内皮细胞增殖率以及附着的HUVECs重要免疫反应的保留,支持了重力纺PCL纤维在血管组织工程三维支架生产中的应用。