Summary
Tissue engineering aims to provide a temporary scaffold for repair at the site of injury or disease that is able to support cell attachment and growth while synthesis of matrix proteins and reorganization take place. Although relatively successful, bladder tissue engineering suffers from the formation of scar tissue at the scaffold implant site partly due to the phenotypic switch of smooth muscle cells (SMCs) from a quiescent contractile phenotype to a synthetic proliferative phenotype, known as myofibroblast. We hypothesize that culturing human SMCs in enzymatically degradable poly(ethylene) glycol (PEG) hydrogels modified with integrin-binding peptides, and in co-culture with human urothelial cells (UCs), will offer some insight as to the required environment for their subsequent differentiation into quiescent SMCs. We have established protocols for isolation, culture, and characterization of human bladder UCs, SMCs, and fibroblasts and investigated co-culture conditions for SMCs and UCs. The optimal PEG hydrogel properties, promoting growth of these cells, have been investigated by varying the amounts of cell adhesion peptide, PEG, and crosslinker and examined using light and fluorescence microscopy. Furthermore, the cell organization within and on top of gels 14 days post seeding has been examined by histology and immunohistochemistry. We have investigated a co-culture model for UCs and SMCs integrated into PEG hydrogels, mimicking a section of the bladder wall for reconstructive purposes that also could contribute to the understanding of the underlying basic mechanisms of SMC differentiation.
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References
Oberpenning, F., Meng, J., Yoo, J. J., and Atala, A. (1999) De novo reconstitution of a functional mammalian urinary bladder by tissue engineering. Nat. Biotechnol. 17, 149–155.
Kim, W. J. H. (2000) Cellular signaling in tissue regeneration. Yonsei Med. J. 41, 692–703.
Badylak, S. F. (2004) Xenogeneic extracellular matrix as a scaffold for tissue reconstruction. Transpl. Immunol. 12, 367–377.
Kanematsu, A., Yamamoto, S., Ozeki, M., Noguchi, T., Kanatani, I., Ogawa, O., and Tabata, Y. (2004) Collagenous matrices as release carriers of exogenous growth factors. Biomaterials 25, 4513–4520.
Cross, W. R., Eardley, I., Leese, H. J., and Southgate, J. (2005) A biomimetic tissue from cultured normal human urothelial cells: analysis of physiological function. Am. J. Physiol. Renal Physiol. 289, F459–F468.
Sutherland, R. S., Baskin, L. S., Hayward, S. W., and Cunha, G. R. (1996) Regeneration of bladder urothelium, smooth muscle, blood vessels and nerves into an acellular tissue matrix. J. Urol. 156, 571–577.
Kropp, B. P., Badylak, S., and Thor, K. B. (1995) Regenerative bladder augmentation: a review of the initial preclinical studies with porcine small intestinal submucosa. Adv. Exp. Med. Biol. 385, 229–235.
Kropp, B. P., Rippy, M. K., Badylak, S. F., Adams, M. C., Keating, M. A., Rink, R. C., and Thor, K. B. (1996) Regenerative urinary bladder augmentation using small intestinal submucosa: urodynamic and histopathologic assessment in long-term canine bladder augmentations. J. Urol. 155, 2098–2104.
Probst, M., Dahiya, R., Carrier, S., and Tanagho, E. A. (1997) Reproduction of functional smooth muscle tissue and partial bladder replacement. Br. J. Urol. 79, 505–515.
Yoo, J. J., Meng, J., Oberpenning, F., and Atala, A. (1998) Bladder augmentation using allogenic bladder submucosa seeded with cells. Urology 51, 221–225.
Lutolf, M. P. and Hubbell, J. A. (2003) Synthesis and physicochemical characterization of end-linked poly(ethylene glycol)-co-peptide hydrogels formed by Michael-type addition. Biomacromolecules 4, 713–722.
Hubschmid, U., Leong-Morgenthaler, P. M., Basset-Dardare, A., Ruault, S., and Frey, P. (2005) In vitro growth of human urinary tract smooth muscle cells on laminin and collagen type I-coated membranes under static and dynamic conditions. Tissue Eng. 11, 161–171.
Southgate, J., Masters, J. R. W., and Trejdosiewicz, L. K. (2002) Culture of human urothelium, in Culture of Epithelial Cells (Freshney, R. I. F. and Freshney, M. G., eds), Wiley-Liss, New York, NY, pp. 381–399.
Sugasi, S., Lesbros, Y., Bisson, I., Zhang, Y. Y., Kucera, P., and Frey, P. (2000) In vitro engineering of human stratified urothelium: analysis of its morphology and function. J. Urol. 164, 51–957.
Gomm, J. J., Browne, P. J., Coope, R. C., Liu, Q. Y., Buluwela, L., and Coombes, R. C. (1995) Isolation of pure populations of epithelial and myoepithelial cells from the normal human mammary gland using immunomagnetic separation with Dynabeads. Anal. Biochem. 226, 91–9.
Lutolf, M. P., Raeber, G. P., Zisch, A. H., Tirelli, N., and Hubbell, J. A. (2003) Cell-responsive synthetic hydrogels. Adv. Mater. 15, 888–892.
Lutolf, M. P., Weber, F. E., Schmoekel, H. G., Schense, J. C., Kohler, T., Muller, R., and Hubbell, J. A. (2003) Repair of bone defects using synthetic mimetics of collagenous extracellular matrices. Nat. Biotechnol. 21, 513–518.
Zisch, A. H., Lutolf, M. P., Ehrbar, M., Raeber, G. P., Rizzi, S. C., Davies, N., Schmokel, H., Bezuidenhout, D., Djonov, V., Zilla, P., and Hubbell, J. A. (2003) Cell-demanded release of VEGF from synthetic, biointeractive cell ingrowth matrices for vascularized tissue growth. FASEB J. 17, 2260–2262.
Zisch, A. H., Lutolf, M. P., and Hubbell, J. A. (2003) Biopolymeric delivery matrices for angiogenic growth factors. Cardiovasc. Pathol. 12, 295–310.
Blindt, R., Krott, N., Hanrath, P., vom Dahl, J., van Eys, G., and Bosserhoff, A. K. (2002) Expression patterns of integrins on quiescent and invasive smooth muscle cells and impact on cell locomotion. J. Mol. Cell. Cardiol. 34, 1633–1644.
Moiseeva, E. P. (2001) Adhesion receptors of vascular smooth muscle cells and their functions. Cardiovasc. Res. 52, 372–386.
Acknowledgments
The authors thank Professor Tatiana Segura (UCLA, CA, USA), Professor Jeffrey Hubbell, and Carolyn Yong (EPFL, Switzerland) for their scientific advice and technical support. This work was supported by the Swiss National Science Foundation NCCR 404640-101113.
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Adelöw, C.A., Frey, P. (2007). Synthetic Hydrogel Matrices for Guided Bladder Tissue Regeneration. In: Hauser, H., Fussenegger, M. (eds) Tissue Engineering. Methods in Molecular Medicine™, vol 140. Humana Press. https://doi.org/10.1007/978-1-59745-443-8_7
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DOI: https://doi.org/10.1007/978-1-59745-443-8_7
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