Department of Urology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, route267, Geert Grooteplein 16, 6525, GA, Nijmegen, The Netherlands.
Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands.
World J Urol. 2017 Oct;35(10):1531-1539. doi: 10.1007/s00345-017-2013-9. Epub 2017 Feb 23.
Pre-conditioning of a cell seeded construct may improve the functional outcome of a tissue engineered construct for augmentation cystoplasty. The precise effects of mechanical stimulation on urinary bladder cells in vitro are not clear. In this study we investigate the effect of a cyclic uniaxial strain culture on urinary bladder cells which were seeded on a type I collagen scaffold.
Isolated porcine smooth muscle cells or urothelial cells were seeded on a type I collagen scaffolds and cultured under static and dynamic conditions. A uniform cyclic uniaxial strain was applied to the seeded scaffold using a Bose Electroforce Bio-Dynamic bioreactor. Cell proliferation rate and phenotype were investigated, including SEM analysis, RT-PCR and immunohistochemistry for α-Smooth muscle actin, calponin-1, desmin and RCK103 expression to determine the effects of mechanical stimulation on both cell types.
Dynamic stimulation of smooth muscle cell seeded constructs resulted in cell alignment and enhanced proliferation rate. Additionally, expression of α-Smooth muscle actin and calponin-1 was increased suggesting differentiation of smooth muscle cells to a more mature phenotype.
Mechanical stimuli did not enhance the proliferation and differentiation of urothelial cells. Mechanical stimulation, i.e., preconditioning may improve the functional in vivo outcome of smooth muscle cell seeded constructs for flexible organs such as the bladder.
在细胞接种构建物之前进行预处理可能会改善用于增强性膀胱成形术的组织工程构建物的功能结果。机械刺激对体外膀胱细胞的确切影响尚不清楚。在这项研究中,我们研究了在 I 型胶原支架上接种的膀胱细胞的循环单轴应变培养的效果。
将分离的猪平滑肌细胞或尿路上皮细胞接种在 I 型胶原支架上,并在静态和动态条件下培养。使用 Bose Electroforce Bio-Dynamic 生物反应器对接种的支架施加均匀的循环单轴应变。研究细胞增殖率和表型,包括 SEM 分析、RT-PCR 和免疫组织化学,以确定 α-平滑肌肌动蛋白、钙调蛋白-1、结蛋白和 RCK103 表达的机械刺激对两种细胞类型的影响。
对平滑肌细胞接种构建物的动态刺激导致细胞排列和增殖率增加。此外,α-平滑肌肌动蛋白和钙调蛋白-1的表达增加,表明平滑肌细胞向更成熟的表型分化。
机械刺激不会增强尿路上皮细胞的增殖和分化。机械刺激,即预处理,可能会改善平滑肌细胞接种构建物在诸如膀胱等灵活器官中的体内功能结果。