Abstract

PurposePre-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.MethodsIsolated 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.ResultsDynamic 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.ConclusionsMechanical 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.

Highlights

  • For patients who need bladder reconstruction, a tissue engineered bladder may be an alternative to current procedures in which autologous bowel tissue is used

  • Cell proliferation and differentiation can be influenced and it may lead to extracellular matrix (ECM) production [4,5,6]

  • Hydrostatic pressure on human bladder smooth muscle cells on aligned nanofibrous scaffolds resulted in functional improvement of the engineered tissue [10]

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Summary

Introduction

For patients who need bladder reconstruction, a tissue engineered bladder may be an alternative to current procedures in which autologous bowel tissue is used. Bladder regeneration studies in animals indicated slow smooth muscle cell ingrowth in scaffolds, probably due to the limited migration from adjacent tissue [2] This suggests that adequate conditioning of the tissue engineered construct may be needed to improve the functional outcome of the regenerated tissue for flexible organs. Hydrostatic pressure on human bladder smooth muscle cells on aligned nanofibrous scaffolds resulted in functional improvement of the engineered tissue [10]. This setting simulated in vivo conditions, the exact impact of the mechanical stimulation on the urinary bladder cells is not clear. In this study we investigated the effect of a long-term controlled cyclic uniaxial strain on urinary bladder cells which were seeded on a type I collagen scaffold mimicking the filling and emptying of the bladder to assess whether this pre-conditioning step is beneficial in urinary bladder tissue engineering

Materials and methods
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Results
Evaluation of bioreactor cultured scaffolds
Discussion
Compliance with ethical standards
Full Text
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