Abstract
Purpose
The purpose of this study was to describe and test a kind of stretch pattern which is based on modified BOSE BioDynamic system to produce optimum physiological stretch during bladder cycle. Moreover, we aimed to emphasize the effects of physiological stretch’s amplitude upon proliferation and contractility of human bladder smooth muscle cells (HBSMCs).
Methods
HBSMCs were seeded onto silicone membrane and subjected to stretch simulating bladder cycle at the range of stretches and time according to customized software on modified BOSE BioDynamic bioreactor. Morphological changes were assessed using immunofluorescence and confocal laser scanning microscope. Cell proliferation and cell viability were determined by BrdU incorporation assay and Cell Counting Kit-8, respectively. Contractility of the cells was determined using collagen gel contraction assay. RT-PCR was used to assess phenotypic and contractility markers.
Results
HBSMCs were found to show morphologically spindle-shaped and orientation at various elongations in the modified bioreactor. Stretch-induced proliferation and viability depended on the magnitude of stretch, and stretches also regulate contractility and contraction markers in a magnitude-dependent manner.
Conclusion
We described and tested a kind of stretch pattern which delivers physiological stretch implemented during bladder cycle. The findings also showed that mechanical stretch can promote magnitude-dependent morphological, proliferative and contractile modulation of HBSMCs in vitro.
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References
Gurocak S, Nuininga J, Ure I, De Gier RP, Tan MO, Feitz W (2007) Bladder augmentation: review of the literature and recent advances. Indian J Urol 23(4):452–457. doi:10.4103/0970-1591.36721
Atala A (2011) Tissue engineering of human bladder. Br Med Bull 97:81–104. doi:10.1093/bmb/ldr003
Robert L (2002) Preface. In: Atala A, Lanza RP (eds) Methods of tissue engineering, 1st edn. Academic Press, San Diego, pp xv–xvii
Atala A, Bauer SB, Soker S, Yoo JJ, Retik AB (2006) Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet 367(9518):1241–1246. doi:10.1016/S0140-6736(06)68438-9
Atala A (2009) Regenerative medicine and tissue engineering in urology. Urol Clin N Am 36(2):199–209. doi:10.1016/j.ucl.2009.02.009
Atala A, Koh C (2005) Applications of tissue engineering in the genitourinary tract. Expert Rev Med Devices 2(1):119–126. doi:10.1586/17434440.2.1.119
Ram-Liebig G, Ravens U, Balana B, Haase M, Baretton G, Wirth MP (2006) New approaches in the modulation of bladder smooth muscle cells on viable detrusor constructs. World J Urol 24(4):429–437. doi:10.1007/s00345-006-0104-0
Chen L, Wei TQ, Wang Y, Zhang J, Li H, Wang KJ (2012) Simulated bladder pressure stimulates human bladder smooth muscle cell proliferation via the PI3K/SGK1 signaling pathway. J Urol 188(2):661–667. doi:10.1016/j.juro.2012.03.112
Ngo P, Ramalingam P, Phillips JA, Furuta GT (2006) Collagen gel contraction assay. Methods Mol Biol 341:103–109. doi:10.1385/1-59745-113-4:103
Leung DY, Glagov S, Mathews MB (1976) Cyclic stretching stimulates synthesis of matrix components by arterial smooth muscle cells in vitro. Science 191(4226):475–477
Geest JPV, Di Martino ES, Vorp DA (2004) An analysis of the complete strain field within Flexercell membranes. J Biomech 37(12):1923–1928
Halachmi S, Aitken KJ, Szybowska M, Sabha N, Dessouki S, Lorenzo A, Tse D, Bagli DJ (2006) Role of signal transducer and activator of transcription 3 (STAT3) in stretch injury to bladder smooth muscle cells. Cell Tissue Res 326(1):149–158. doi:10.1007/s00441-006-0204-6
Bouhout S, Gauvin R, Gibot L, Aube D, Bolduc S (2011) Bladder substitute reconstructed in a physiological pressure environment. J Pediatr Urol 7(3):276–282. doi:10.1016/j.jpurol.2011.03.002
Qu MJ, Liu B, Wang HQ, Yan ZQ, Shen BR, Jiang ZL (2007) Frequency-dependent phenotype modulation of vascular smooth muscle cells under cyclic mechanical strain. J Vasc Res 44(5):345–353. doi:10.1159/000102278
Houtchens GR, Foster MD, Desai TA, Morgan EF, Wong JY (2008) Combined effects of microtopography and cyclic strain on vascular smooth muscle cell orientation. J Biomech 41(4):762–769. doi:10.1016/j.jbiomech.2007.11.027
Pang Y, Wang X, Lee D, Greisler HP (2011) Dynamic quantitative visualization of single cell alignment and migration and matrix remodeling in 3-D collagen hydrogels under mechanical force. Biomaterials 32(15):3776–3783. doi:10.1016/j.biomaterials.2011.02.003
Adam RMRJ, Cheng HL, Rice DC, Khoury J, Bauer SB, Peters CA, Freeman MR (2003) Signaling through PI3K/Akt mediates stretch and PDGF-BB-dependent DNA synthesis in bladder smooth muscle cells. J Urol 169(6):2388–2393
Orsola A, Adam RM, Peters CA, Freeman MR (2002) The decision to undergo DNA or protein synthesis is determined by the degree of mechanical deformation in human bladder muscle cells. Urology 59(5):779–783
Upadhyay J, Aitken KJ, Damdar C, Bolduc S, Bagli DJ (2003) Integrins expressed with bladder extracellular matrix after stretch injury in vivo mediate bladder smooth muscle cell growth in vitro. J Urol 169(2):750–755. doi:10.1097/01.ju.0000051682.61041.a5
Baskin L, Meaney D, Landsman A, Zderic SA, Macarak E (1994) Bovine bladder compliance increases with normal fetal development. J Urol 152(2 Pt 2):692–695; discussion 696-697
DE Coplen ME, Howard PS (2003) Matrix synthesis by bladder smooth muscle cells is modulated by stretch frequency. Vitro Cell Dev Biol Anim 39(3–4):157–162
Kanda K, Matsuda T, Oka T (1992) Two-dimensional orientational response of smooth muscle cells to cyclic stretching. ASAIO J 38(3):M382–M385
Birukov KG, Shirinsky VP, Stepanova OV, Tkachuk VA, Hahn AW, Resink TJ, Smirnov VN (1995) Stretch affects phenotype and proliferation of vascular smooth muscle cells. Mol Cell Biochem 144(2):131–139
Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (No. 81300579, 30872593, 31170907 and 81100494), the Technology Support Program of Science and Technology Department of Sichuan Province (No. 2010SZ0163) and the Ph.D. Programs Foundation of Ministry of Education of China (No. 20110181110028).
Conflict of interest
The manuscript does not contain clinical studies or patient data. The authors declare that they have no conflict of interest.
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De-Yi Luo and Romel Wazir contributed equally to this work and should be considered co-first authors.
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Luo, DY., Wazir, R., Du, C. et al. Magnitude-dependent proliferation and contractility modulation of human bladder smooth muscle cells under physiological stretch. World J Urol 33, 1881–1887 (2015). https://doi.org/10.1007/s00345-015-1509-4
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DOI: https://doi.org/10.1007/s00345-015-1509-4