Stankus John J, Guan Jianjun, Fujimoto Kazuro, Wagner William R
Department of Chemical Engineering, 100 Technology Drive, University of Pittsburgh, PA 15261, USA.
Biomaterials. 2006 Feb;27(5):735-44. doi: 10.1016/j.biomaterials.2005.06.020. Epub 2005 Aug 10.
Electrospinning permits fabrication of biodegradable elastomers into matrices that can resemble the scale and mechanical behavior of the native extracellular matrix. However, achieving high-cellular density and infiltration with this technique remains challenging and time consuming. We have overcome this limitation by electrospraying vascular smooth muscle cells (SMCs) concurrently with electrospinning a biodegradable, elastomeric poly(ester urethane)urea (PEUU). Trypan blue staining revealed no significant decrease in cell viability from the fabrication process and electrosprayed SMCs spread and proliferated similar to control unprocessed SMCs. The resulting SMC microintegrated PEUU constructs were cultured under static conditions or transmural perfusion. Higher cell numbers resulted with perfusion culture with 131% and 98% more viable cells versus static culture at days 4 and 7 (p<0.05). Fluorescent imaging and hematoxylin and eosin staining further illustrated high cell densities integrated between the elastomeric fibers after perfusion culture. SMC microintegrated PEUU was strong, flexible and anisotropic with tensile strengths ranging from 2.0 to 6.5 MPa and breaking strains from 850 to 1,700% dependent on the material axis. The ability to microintegrate smooth muscle or other cell types into a biodegradable elastomer fiber matrix embodies a novel tissue engineering approach that could be applied to fabricate high cell density elastic tissue mimetics, blood vessels or other cardiovascular tissues.
静电纺丝技术可将可生物降解的弹性体制成类似于天然细胞外基质的尺寸和力学行为的基质。然而,利用该技术实现高细胞密度和细胞浸润仍然具有挑战性且耗时。我们通过在静电纺丝可生物降解的弹性聚(酯脲)脲(PEUU)的同时电喷雾血管平滑肌细胞(SMC)克服了这一限制。台盼蓝染色显示,制备过程对细胞活力没有显著降低,且电喷雾的SMC与未处理的对照SMC一样能够铺展和增殖。将所得的SMC微整合PEUU构建体在静态条件下或经壁灌注培养。在第4天和第7天,灌注培养产生的细胞数量更多,存活细胞比静态培养分别多131%和98%(p<0.05)。荧光成像以及苏木精和伊红染色进一步表明,灌注培养后弹性纤维之间整合了高细胞密度。SMC微整合PEUU坚固、柔韧且具有各向异性,其拉伸强度范围为2.0至6.5MPa,断裂应变在850%至1700%之间,具体取决于材料轴。将平滑肌或其他细胞类型微整合到可生物降解的弹性体纤维基质中的能力体现了一种新型的组织工程方法,可应用于制造高细胞密度的弹性组织模拟物、血管或其他心血管组织。