Bladder acellular matrix as a substrate for studying in vitro bladder smooth muscle-urothelial cell interactions.

The objective of this study was to evaluate the ability of bladder acellular matrix (BAM) to support the individual and combined growth of primary porcine bladder smooth muscle (SMC) and urothelial (UEC) cells. An in vitro co-culture system was devised to evaluate the effect of UEC on (i) SMC-mediated contraction of BAM discs, and (ii) SMC invasiveness into BAM. Cells were seeded onto BAM discs under 4 different culture conditions. Constructs were incubated for 1, 7, 14 and 28 days. Samples were then harvested for evaluation of matrix contraction. Immunohistochemistry (IHC) was utilized to examine cellular organization within the samples and conditioned media supernatants analyzed for net gelatinase activity. BAM contraction was significantly increased with co-culture. The same side co-culture configuration lead to a greater reduction in surface area than opposite side co-culture. IHC revealed enhanced SMC infiltration into BAM when co-culture was utilized. A significant increase in net gelatinase activity was also observed with the co-culture configuration. Enhanced infiltration and contractile ability of bladder SMCs with UEC co-culture may, in part, be due to an increase in gelatinase activity. The influence of bladder UECs on SMC behaviour in vitro indicates that BAM may contain some key inductive factors that serve to promote important bladder cell-cell and cell-matrix interactions.

[1]  P. Friedl,et al.  The biology of cell locomotion within three-dimensional extracellular matrix , 2000, Cellular and Molecular Life Sciences CMLS.

[2]  K. M. Haberstroh,et al.  Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O. , 2002, The Journal of urology.

[3]  E. Elson,et al.  Reciprocal interactions between cells and extracellular matrix during remodeling of tissue constructs. , 2002, Biophysical chemistry.

[4]  P. Frey,et al.  Coculture of bladder urothelial and smooth muscle cells on small intestinal submucosa: potential applications for tissue engineering technology. , 2000, The Journal of urology.

[5]  S. Badylak,et al.  Small Intestinal Submucosa as an Intra-Articular Ligamentous Graft Material: A Pilot Study in Dogs , 1994, Phlebologie.

[6]  A. Shokeir.  Bladder regeneration: between the idea and reality , 2002, BJU international.

[7]  J. Squire,et al.  A new twist in the collagen story—the type VI segmented supercoil , 2001, The EMBO journal.

[8]  P. Merguerian,et al.  Regeneration of functional bladder substitutes using large segment acellular matrix allografts in a porcine model. , 2000, The Journal of urology.

[9]  S. Hayward,et al.  Mesenchymal-epithelial interactions in bladder smooth muscle development: epithelial specificity. , 1998, The Journal of urology.

[10]  E. Lakatta,et al.  Migration of cultured vascular smooth muscle cells through a basement membrane barrier requires type IV collagenase activity and is inhibited by cellular differentiation. , 1994, Circulation research.

[11]  P. Howard,et al.  Effect of physical forces on bladder smooth muscle and urothelium. , 1993, The Journal of urology.

[12]  L. McCulloch,et al.  THE HYALURONIC ACID RECEPTORS INDUCED BY STRETCH INJURY OF RAT BLADDER IN VIVO AND INFLUENCES SMOOTH MUSCLE CELL CONTRACTION IN VITRO , 1999 .

[13]  J. Southgate,et al.  The role of matrix metalloproteinases in an in vitro model of bladder tumor invasion. , 1999, Advances in experimental medicine and biology.

[14]  R. T. Lee,et al.  Integrin-mediated collagen matrix reorganization by cultured human vascular smooth muscle cells. , 1995, Circulation research.

[15]  D. F. Thomas,et al.  Urothelial tissue culture for bladder reconstruction: an experimental study. , 1993, The Journal of urology.

[16]  R. Timpl,et al.  Expression of type VI collagen mRNA during wound healing. , 1993, The Journal of investigative dermatology.

[17]  D. Schuppan,et al.  Collagen VI regulates normal and transformed mesenchymal cell proliferation in vitro. , 1996, Experimental cell research.

[18]  P. Frey,et al.  Expansion and long-term culture of differentiated normal rat urothelial cells in vitro , 2001, In Vitro Cellular & Developmental Biology - Animal.

[19]  M. van Griensven,et al.  Immunohistochemical localization of collagen VI in arthrofibrosis , 1999, Archives of Orthopaedic and Trauma Surgery.

[20]  L A Geddes,et al.  Small intestinal submucosa as a vascular graft: a review. , 1993, Journal of investigative surgery : the official journal of the Academy of Surgical Research.

[21]  G. Fields,et al.  Matrix metalloproteinases and collagen catabolism. , 2002, Biopolymers.

[22]  D. Wiederschain,et al.  Dysregulated proteolytic balance as the basis of excess extracellular matrix in fibrotic disease. , 1997, The American journal of physiology.

[23]  N. Carragher,et al.  Degraded Collagen Fragments Promote Rapid Disassembly of Smooth Muscle Focal Adhesions That Correlates with Cleavage of Pp125FAK, Paxillin, and Talin , 1999, The Journal of cell biology.

[24]  R. Tarnuzzer,et al.  Matrix metalloproteinase inhibition modulates fibroblast-mediated matrix contraction and collagen production in vitro. , 2003, Investigative ophthalmology & visual science.

[25]  S. Hayward,et al.  Diffusable growth factors induce bladder smooth muscle differentiation , 2000, In Vitro Cellular & Developmental Biology - Animal.

[26]  Kenneth M. Yamada,et al.  Transmembrane crosstalk between the extracellular matrix and the cytoskeleton , 2001, Nature Reviews Molecular Cell Biology.

[27]  K. Porter,et al.  Enhanced invasive properties exhibited by smooth muscle cells are associated with elevated production of MMP-2 in patients with aortic aneurysms. , 2002, European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery.

[28]  T. K. Hunt,et al.  Stromelysin-1-deficient fibroblasts display impaired contraction in vitro. , 1999, The Journal of surgical research.

[29]  H. Miyake,et al.  Significance of matrix metalloproteinases and tissue inhibitors of metalloproteinase expression in the recurrence of superficial transitional cell carcinoma of the bladder. , 2001, Journal of Urology.

[30]  F. Grinnell,et al.  Differences in the Regulation of Fibroblast Contraction of Floating Versus Stressed Collagen Matrices* , 1999, The Journal of Biological Chemistry.

[31]  C. Kielty,et al.  Attachment of human vascular smooth muscles cells to intact microfibrillar assemblies of collagen VI and fibrillin. , 1992, Journal of cell science.

[32]  I. Grierson,et al.  Matrix metalloproteinases: a role in the contraction of vitreo-retinal scar tissue. , 2001, The American journal of pathology.

[33]  A. Clowes,et al.  Matrix metalloproteinase-9 overexpression enhances vascular smooth muscle cell migration and alters remodeling in the injured rat carotid artery. , 1999, Circulation research.

[34]  A. Brading,et al.  Smooth muscle of the bladder in the normal and the diseased state: pathophysiology, diagnosis and treatment. , 1997, Pharmacology & therapeutics.

[35]  W. Stetler-Stevenson,et al.  Primate smooth muscle cell migration from aortic explants is mediated by endogenous platelet-derived growth factor and basic fibroblast growth factor acting through matrix metalloproteinases 2 and 9. , 1997, Circulation.

[36]  S. Badylak,et al.  Characterization of small intestinal submucosa regenerated canine detrusor: assessment of reinnervation, in vitro compliance and contractility. , 1996, The Journal of urology.

[37]  R. Dahiya,et al.  Homologous bladder augmentation in dog with the bladder acellular matrix graft , 2000, BJU international.

[38]  P. Merguerian,et al.  22 week assessment of bladder acellular matrix as a bladder augmentation material in a porcine model. , 2002, Biomaterials.

[39]  S. Kamidono,et al.  Bladder reconstruction with autotransplanted ileum in the dog: better functional results than standard enterocystoplasty , 2001, BJU international.

[40]  N. Maitland,et al.  Experimental prostate epithelial morphogenesis in response to stroma and three-dimensional matrigel culture. , 2001, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[41]  R. Fässler,et al.  Insights into extracellular matrix functions from mutant mouse models. , 2000, Experimental cell research.

[42]  Frederick Grinnell,et al.  Fibroblast biology in three-dimensional collagen matrices. , 2003, Trends in cell biology.

[43]  Kenneth M. Yamada,et al.  Taking Cell-Matrix Adhesions to the Third Dimension , 2001, Science.

[44]  S. Badylak,et al.  Small Intestinal Submucosa: Utilization as a Wound Dressing in Full‐Thickness Rodent Wounds , 1995, Annals of plastic surgery.

[45]  S. Badylak,et al.  Small intestinal submucosa: a rapidly resorbed bioscaffold for augmentation cystoplasty in a dog model. , 1998, Tissue engineering.

[46]  Stephen F Badylak,et al.  The extracellular matrix as a scaffold for tissue reconstruction. , 2002, Seminars in cell & developmental biology.

[47]  T. Kälble,et al.  Metabolic and functional consequences of urinary reconstruction with bowel , 2003, BJU international.

[48]  T. Schoeller,et al.  Gracilis muscle flap with a tissue‐engineered lining for experimental bladder wall reconstruction , 2001, BJU international.

[49]  D. Stocum.  Regenerative biology: a millenial revolution. , 1999, Seminars in cell & developmental biology.

[50]  T. Krummel,et al.  Matrix metalloproteinases and the ontogeny of scarless repair: the other side of the wound healing balance. , 2002, Plastic and reconstructive surgery.

[51]  D. Mosher,et al.  Contraction of collagen matrices mediated by alpha2beta1A and alpha(v)beta3 integrins. , 2000, Journal of cell science.

[52]  J. Lunec,et al.  Matrix metalloproteinases (MMPs) in bladder cancer: the induction of MMP9 by epidermal growth factor and its detection in urine , 2003, BJU international.

[53]  P. Howard,et al.  Roles of the lamina propria and the detrusor in tension transfer during bladder filling. , 1999, Scandinavian journal of urology and nephrology. Supplementum.

[54]  D. F. Thomas,et al.  Normal human urothelial cells in vitro: proliferation and induction of stratification. , 1994, Laboratory investigation; a journal of technical methods and pathology.

[55]  J. Werkmeister,et al.  Monoclonal antibodies to type VI collagen demonstrate new tissue augmentation of a collagen-based biomaterial implant. , 1993, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.