Laboratory of Membrane Biophysics and Macromolecules, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran.
J Mater Sci Mater Med. 2012 Sep;23(9):2281-90. doi: 10.1007/s10856-012-4688-1. Epub 2012 Jun 7.
Bladder tissue engineering has been the focus of many studies due to its highly therapeutic potential. In this regard many aspects such as biochemical and biomechanical factors need to be studied extensively. Mechanical stimulations such as hydrostatic pressure and topology of the matrices are critical features which affect the normal functions of cells involved in bladder regeneration. In this study, hydrostatic pressure (10 cm H(2)O) and stretch forces were exerted on human bladder smooth muscle cells (hBSMCs) seeded on aligned nanofibrous polycaprolactone/PLLA scaffolds, and the alterations in gene and protein expressions were studied. The gene transcription patterns for collagen type I, III, IV, elastin, α-SMA, calponin and caldesmon were monitored on days 3 and 5 quantitatively. Changes in the expressions of α-SMA, desmin, collagen type I and III were quantified by Enzyme-linked immuno-sorbent assay. The scaffolds were characterized using scanning electron microscope, contact angle measurement and tensile testing. The positive effect of mechanical forces on the functional improvement of the engineered tissue was supported by translational down-regulation of α-SMA and VWF, up-regulation of desmin and improvement of collagen type III:I ratio. Altogether, our study reveals that proper hydrostatic pressure in combination with appropriate surface stimulation on hBSMCs causes a tissue-specific phenotype that needs to be considered in bladder tissue engineering.
膀胱组织工程一直是许多研究的焦点,因为它具有很高的治疗潜力。在这方面,需要广泛研究生物化学和生物力学等方面。水静压和基质拓扑等机械刺激是影响膀胱再生中细胞正常功能的关键特征。在这项研究中,对种植在取向的纳米纤维聚己内酯/聚丙交酯支架上的人膀胱平滑肌细胞(hBSMC)施加水静压(10cmH2O)和拉伸力,并研究了基因和蛋白质表达的变化。在第 3 天和第 5 天定量监测了胶原蛋白 I、III、IV、弹性蛋白、α-SMA、钙调蛋白和钙调蛋白基因的转录模式。通过酶联免疫吸附试验定量检测 α-SMA、结蛋白、胶原蛋白 I 和 III 的表达变化。使用扫描电子显微镜、接触角测量和拉伸试验对支架进行了表征。研究结果表明,机械力对工程组织功能的改善有积极作用,其作用机制是下调 α-SMA 和 VWF 的表达,上调结蛋白的表达,并改善 III 型胶原蛋白与 I 型胶原蛋白的比值。总的来说,我们的研究表明,适当的水静压结合适当的 hBSMC 表面刺激会导致组织特异性表型,这在膀胱组织工程中需要加以考虑。