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Rapid differentiation of superficial urothelial cells after chitosan-induced desquamation

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Abstract

Superficial cell desquamation followed by differentiation of newly exposed superficial cells induces regeneration of the urinary bladder epithelium, urothelium. In the present work, chitosan was evaluated as a new inducer of urothelial cell desquamation, in order to study the regeneration of mouse urothelial cells in vivo. Intravesical application of chitosan dispersion caused complete removal of only the superficial layer of cells within 20 min of treatment. Differentiation of the new superficial layer was followed by the appearance and distribution of three urothelial differentiation markers, tight junction protein ZO1, cytokeratin 20 and the maturation of the apical plasma membrane. The arrangement of ZO1 into continuous lines in individual cells of the intermediate layer was already found after 10 min of chitosan application, when desquamation had just started. The appearance of the apical membrane changed from microvillar to typically scalloped within 20 min of regeneration, while complete arrangement of the cytokeratin 20 network took 60 min. These findings provide a new perspective on the rate of the differentiation process in the urothelium and make chitosan a new and a very controllable tool for studies on urothelial regeneration.

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References

  • Acharya P, Beckel J, Ruiz WG, Wang E, Rojas R, Birder L, Apodaca G (2004) Distribution of the tight junction proteins ZO-1, occludin, and claudin-4, -8, and -12 in bladder epithelium. Am J Physiol Renal Physiol 287:F305–F318

    Article  PubMed  CAS  Google Scholar 

  • Aronson M, Medalia O, Amichay D, Nativ O (1988) Endotoxin-induced shedding of viable uroepithelial cells is an antimicrobial defense mechanism. Infect Immun 56:1615–1617

    PubMed  CAS  Google Scholar 

  • Artursson P, Lindmark T, Davis SS, Illum L (1994) Effect of chitosan on the permeability of monolayers of intestinal epithelial cells (Caco-2). Pharm Res 11:1358–1361

    Article  PubMed  CAS  Google Scholar 

  • Baskin L, Meaney D, Landsman A, Zderic SA, Macarak E (1994) Bovine bladder compliance increases with normal fetal development. J Urol 152:692–695

    PubMed  CAS  Google Scholar 

  • Baskin LS, Hayward SW, Young PF, Cunha GR (1996) Ontogeny of the rat bladder: smooth muscle and epithelial differentiation. Acta Anat (Basel) 155:163–171

    Article  CAS  Google Scholar 

  • Dalal E, Medalia O, Harari O, Aronson M (1994) Moderate stress protects female mice against bacterial infection of the bladder by eliciting uroepithelial shedding. Infect Immun 62:5505–5510

    PubMed  CAS  Google Scholar 

  • Erman A, Jezernik K, Stiblar-Martincic D, Romih R, Veranic P (2001) Postnatal restoration of the mouse urinary bladder urothelium. Histochem Cell Biol 115:309–316

    PubMed  CAS  Google Scholar 

  • Farsund T (1975) Cell kinetics of mouse urinary bladder epithelium. I. Circadian and age variations in cell proliferation and nuclear DNA content. Virchows Arch B Cell Pathol 18:35–49

    PubMed  CAS  Google Scholar 

  • Goldstein P, Kroemer G (2006) Cell death by necrosis: towards a molecular definition. TRENDS Biochem Sci 32:37–43

    Article  Google Scholar 

  • Hasegawa M, Yagi K, Iwakawa S, Hirai M (2001) Chitosan induces apoptosis via caspase-3 activation in bladder tumor cells. Jpn J Cancer Res 92:459–466

    PubMed  CAS  Google Scholar 

  • Hejazi R, Amiji M (2003) Chitosan-based gastrointestinal delivery systems. J Control Release 89:151–165

    Article  PubMed  CAS  Google Scholar 

  • Hu P, Meyers S, Liang FX, Deng FM, Kachar B, Zeidel ML, Sun TT (2002) Role of membrane proteins in permeability barrier function: uroplakin ablation elevates urothelial permeability. Am J Physiol Renal Physiol 283:F1200–F1207

    PubMed  CAS  Google Scholar 

  • Jacob J, Hindmarsh JR, Ludgate CM, Chisholm GD (1982) Observations on the ultrastructure of human urothelium: the response of normal bladder of elderly subjects to hyperthermia. Urol Res 10:227–237

    Article  PubMed  CAS  Google Scholar 

  • Jenings CD, Boleyn K, Bridges SR, Wood PJ, Anderson JW (1988) A comparison of the lipid-lowering and intestinal morphological effects of cholestyramine, chitosan, and oat gum in rats. Proc Soc Exp Biol Med 189:13–20

    Google Scholar 

  • Jost SP (1989) Cell cycle of normal bladder urothelium in developing and adult mice. Virchows Arch B Cell Pathol Incl Mol Pathol 57:27–36

    Article  PubMed  CAS  Google Scholar 

  • Kachar B, Liang F, Lins U, Ding M, Wu XR, Stoffler D, Aebi U, Sun TT (1999) Three-dimensional analysis of the 16 nm urothelial plaque particle: luminal surface exposure, preferential head-to-head interaction, and hinge formation. J Mol Biol 15:595–608

    Article  Google Scholar 

  • Kerec M, Bogataj M, Veranic P, Mrhar A (2005) Permeability of pig urinary bladder wall: the effect of chitosan and the role of calcium. Eur J Pharm Sci 25:113–121

    Article  PubMed  CAS  Google Scholar 

  • Kos MK, Bogataj M, Veranic P, Mrhar A (2006) Permeability of pig urinary bladder wall: time and concentration dependent effect of chitosan. Biol Pharm Bull 29:1685–1691

    Article  PubMed  CAS  Google Scholar 

  • Koss LG (1967) A light and electron microscopic study of the effects of a single dose of cyclophosphamide on various organs in the rat. I. The urinary bladder. Lab Invest 16:44–65

    PubMed  CAS  Google Scholar 

  • Kotzé AF, Luessen HL, de Leeuw BJ, de Boer BG, Verhoef JC, Junginger HE (1997) N-Trimethyl chitosan chloride as a potential absorption enhancer across mucosal surfaces: in vitro evaluation in intestinal epithelial cells (Caco-2). Pharm Res 14:1197–1202

    Article  PubMed  Google Scholar 

  • Kreft ME, Sterle M, Veranic P, Jezernik K (2005) Urothelial injuries and the early wound healing response: tight junctions and urothelial cytodifferentiation. Histochem Cell Biol 123:529–539

    Article  PubMed  CAS  Google Scholar 

  • Kunze E, Wöltjen HH, Nehm FJ, Schauer A (1981) Cell cycle kinetics of regenerating urothelial cells of the rat urinary bladder after partial cystectomy. Virchows Arch B Cell Pathol Incl Mol Pathol 38:117–125

    Article  PubMed  CAS  Google Scholar 

  • Levi PE, Cooper EH, Anderson CK, Williams RE (1969) Analyses of DNA content, nuclear size and cell proliferation of transitional cell carcinoma in man. Cancer 23:1074–1085

    Article  PubMed  CAS  Google Scholar 

  • Locher GW, Cooper EH (1970) Repair of rat urinary bladder epithelium following injury by cyclophosphamide. Invest Urol 8:116–123

    PubMed  CAS  Google Scholar 

  • Luna EJ, Hitt AL (1992) Cytoskeleton-plasma membrane interactions. Science 258:955–964 Review

    Article  PubMed  CAS  Google Scholar 

  • Messier B, Leblond CP (1960) Cell proliferation and migration as revealed by adioautography after injection of thymidine-H3 into male rats and mice. Am J Anat 106:247–285

    Article  PubMed  CAS  Google Scholar 

  • Miura T, Usami M, Tsuura Y, Ishida H, Seino Y (1995) Hypoglycemic and hypolipidemic effect of chitosan in normal and neonatal streptozotocin-induced diabetic mice. Biol Pharm Bull 18:1623–1625

    PubMed  CAS  Google Scholar 

  • Moll R, Wu XR, Lin JH, Sun TT (1995) Uroplakins, specific membrane proteins of urothelial umbrella cells, as histological markers of metastatic transitional cell carcinomas. Am J Pathol 147:1383–1397

    PubMed  CAS  Google Scholar 

  • Mulvey MA, Lopez-Boado YS, Wilson CL, Roth R, Parks WC, Heuser J, Hultgren SJ (1998) Induction and evasion of host defenses by type 1-piliated uropathogenic Escherichia coli. Science 282:1494–1497

    Article  PubMed  CAS  Google Scholar 

  • Mulvey MA, Schilling JD, Hultgren SJ (2001) Establishment of a persistent Escherichia coli reservoir during the acute phase of a bladder infection. Infect Immun 69:4572–4579

    Article  PubMed  CAS  Google Scholar 

  • Nativ O, Dalal E, Hidas G, Aronson M (2006) EDTA-induced urothelial cell shedding for the treatment of superficial bladder cancer in the mouse. Int J Urol 13:1344–1346

    Article  PubMed  Google Scholar 

  • Philips FS, Sternberg SS, Cronin AP, Vidal PM (1961) Cyclophosphamide and urinary bladder toxicity. Cancer Res 21:1577–1589

    PubMed  CAS  Google Scholar 

  • Ranaldi G, Marigliano I, Vespignani I, Perozzi G, Sambuy Y (2002) The effect of chitosan and other polycations on tight junction permeability in the human intestinal Caco-2 cell line. J Nutr Biochem 13:157–167

    Article  PubMed  CAS  Google Scholar 

  • Romih R, Jezernik K, Masera A (1998) Uroplakins and cytokeratins in the regenerating rat urothelium after sodium saccharin treatment. Histochem Cell Biol 109:263–269

    Article  PubMed  CAS  Google Scholar 

  • Romih R, Veranic P, Jezernik K (1999) Actin filaments during terminal differentiation of urothelial cells in the rat urinary bladder. Histochem Cell Biol 112:375–380

    Article  PubMed  CAS  Google Scholar 

  • Romih R, Koprivec D, Martincic DS, Jezernik K (2001) Restoration of the rat urothelium after cyclophosphamide treatment. Cell Biol Int 25:531–537

    Article  PubMed  CAS  Google Scholar 

  • Şenel S, Hincal AA (2001) Drug permeation enhancement via buccal route: possibilities and limitations. J Control Release 72:133–144

    Article  PubMed  Google Scholar 

  • Stewart FA (1986) Mechanism of bladder damage and repair after treatment with radiation and cytostatic drugs. Br J Cancer Suppl 7:280–291

    PubMed  CAS  Google Scholar 

  • Truschel ST, Wang E, Ruiz WG, Leung SM, Rojas R, Lavelle J, Zeidel M, Stoffer D, Apodaca G (2002) Stretch-regulated exocytosis/endocytosis in bladder umbrella cells. Mol Biol Cell 13:830–846

    Article  PubMed  CAS  Google Scholar 

  • Veranic P, Jezernik K (2000) The response of junctional complexes to induced desquamation in mouse bladder urothelium. Biol Cell 92:105–113

    Article  PubMed  CAS  Google Scholar 

  • Veranic P, Jezernik K (2002) Trajectorial organisation of cytokeratins within the subapical region of umbrella cells. Cell Motil Cytoskeleton 53:317–325

    Article  PubMed  Google Scholar 

  • Veranic P, Romih R, Jezernik K (2004) What determines differentiation of urothelial umbrella cells? Eur J Cell Biol 83:27–34

    Article  PubMed  Google Scholar 

  • Wang Y, Chen M, Wolosin JM (1993) ZO-1 in corneal epithelium; stratal distribution and synthesis induction by outer cell removal. Exp Eye Res 57:283–292

    Article  PubMed  CAS  Google Scholar 

  • Watanabe S, Sasaki J (1992) 12-O-Tetradecanoylphorbol-13-acetate induces selective desquamation of superficial cells in rat urinary bladder epithelium. Cell Tissue Res 268:239–245

    Article  PubMed  CAS  Google Scholar 

  • Wolosin JM, Chen M (1993) Ontogeny of corneal epithelial tight junctions: stratal locale of biosynthetic activities. Invest Ophthalmol Vis Sci 34:2655–2664

    PubMed  CAS  Google Scholar 

  • Yu S, Zhao Y, Wu F, Zhang X, Lu W, Zhang H, Zhang Q (2004) Nasal insulin delivery in the chitosan solution: in vitro and in vivo studies. Int J Pharm 281:11–23

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

The authors thank Mr Martin Cregeen for checking the English of the text.

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Authors and Affiliations

  1. Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Lipičeva 2, Ljubljana, Slovenia

    Peter Veranič, Andreja Erman & Kristijan Jezernik

  2. Faculty of Pharmacy, University of Ljubljana, Aškerčeva 7, 1000, Ljubljana, Slovenia

    Mojca Kerec-Kos, Marija Bogataj & Aleš Mrhar

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  1. Peter Veranič
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  2. Andreja Erman
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  3. Mojca Kerec-Kos
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  4. Marija Bogataj
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  6. Kristijan Jezernik
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Correspondence to Peter Veranič.

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Veranič, P., Erman, A., Kerec-Kos, M. et al. Rapid differentiation of superficial urothelial cells after chitosan-induced desquamation. Histochem Cell Biol 131, 129–139 (2009). https://doi.org/10.1007/s00418-008-0492-x

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