The effect of a cyclic uniaxial strain on urinary bladder cells

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.Electronic supplementary materialThe online version of this article (doi:10.1007/s00345-017-2013-9) contains supplementary material, which is available to authorized users.

Bioengineering Approaches for Bladder Regeneration

“…Natural porous polymer scaffolds can be produced nowadays by many different methods, such as solvent casting, phase separation, gas-foaming techniques, electrospinning, freeze extraction, lyophilization, 3D printing, etc., to produce porous urinary bladder scaffolds with very different degrees of interconnected porosity and with a wide range of pore morphologies [ 34 ]. Thus, an interconnected porous natural type I collagen scaffold with pores ranging from 50 to 100 µm has been recently prepared by lyophilization and utilized as a strip clamped in a Bose Electroforce Bio-Dynamic bioreactor to study the effect of a cyclic uniaxial strain on urinary bladder cells [ 48 ].…”

“…The vascularization of bioengineered organs can be achieved by ex vivo approaches, such as cell sheet engineering, spheroid coculture, scaffold functionalization, modular assembly, and bioprinting or addressed in vivo by arteriovenous loops, polysurgery, and genetic manipulations [ 130 ]. Scaffold survival is profoundly dependent on the formation of a new vascular bed, and this process becomes more difficult with the increase of the construct size [ 48 ]. We believe that the lack of functionality of the engineered bladder tissue is a good reason to discourage researchers and explains somehow the low high-impact scientific production in this scientific area during the last decade (see Table 4 ).…”

“…Finally, one of the most recent contributions to the in vitro bladder bioreactor field studied the effect of a cyclic uniaxial strain on urinary bladder cells seeded on type I collagen scaffold strips [ 48 ], utilizing a BOSE Electroforce BioDynamic ® bioreactor. The dynamic stimulation of the SMC-seeded constructs resulted in cell alignment, enhanced proliferation rate, and differentiation of the SMCs to a more mature phenotype.…”

Bladder Substitution: The Role of Tissue Engineering and Biomaterials

“…The strips were initially seeded statically with SMC or UC and then dynamically in the bioreactor, while control scaffolds were cultured under static conditions. The results demonstrated that conditioning of collagen-based scaffolds by mechanical stimulation leads to more SMC, probably bypassing difficulties related to poor SMC in-growth and muscle development in tissue engineered bladders [67].…”

Isolation of multipotent progenitor cells from pleura and pericardium for tracheal tissue engineering purposes

“…Collagen matrices (type I collagen) were prepared from bovine achilleas tendon (Southern Lights Biomaterials) as previously described. 27 , 28 In brief, a 0,5% (w/v) collagen suspension was made by swelling and subsequently homogenization in 0.25 M acetic acid at 4°C. The suspension was deaerated by centrifugation (at 120 g for 30 min), casted in 6‐well plates and frozen at −20°C.…”