Morphometric and Vascular Analyses and MMP-2 Expression in Bladders of Animals with Bovine Enzootic Haematuria

Anderson Barros Archanjo, Natalia Viana Tamiasso, Juliana de Castro Cosme, Maria Aparecida da Silva, Leonardo Oliveira Trivilin, Julio Lopes Sequeira, Louisiane de Carvalho Nunes

Abstract


Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridium aquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovine papillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognostic markers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a variety of cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantify the expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.

Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigert's resorcin-fuchsin staining methods. The morphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesions were differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation, ectasia, dilation and thickening and the material stained using the Weigert's resorcin-fuchsin method was used to evaluate sclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2 antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia, haemorrhage, cystitis cystica and Brunn's nests and the neoplastic lesions were haemangioma, myxoma, transitional cell carcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differences between the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed the frequent occurrence of different pathological aspects, such as vascular proliferation, dilation, expansion, ectasia and thickening; more severe vascular proliferation, thickening and fibroelastosis were observed in bladders with neoplasms. Metalloproteinase-2 expression was observed in the epithelial cells, fibroblasts, endothelial cells and smooth muscle cells present in the tumour tissue and was a significant increase in the expression of MMP-2 in the neoplasms of mesenchymal origin compared to the neoplasms of epithelial origin.

Discussion: The presence of a large variety of non-neoplastic lesions indicates the possibility that there are other factors concomitant to BEH involved. Epithelial hyperplasia and metaplasia, for example, could be the result of damage caused by bovine papillomavirus type 2 or even an altered urine pH. Furthermore, it is believed that the bladder has the capacity to generate various types of neoplasms due to the different histological components of the bladder, thus enabling it to generate both mesenchymal and epithelial lesions. The thickness of the urothelium was greater in all the quadrants with neoplasms compared to the quadrants without neoplasms. These data revealed that it is possible to use less invasive diagnostic imaging methods to characterise bovine enzootic haematuria and to measure tumour progression based on the severity of vascular changes. The vascular proliferation and thickening were significant in bladders with neoplasms, which may be explained by the fact that tumours require an extensive vascular supply. In addition, many tumours release vascular growth factors and promote neovascularisation. The intensity of the immunostaining of metalloproteinase-2 differed among the different types of neoplasms. However, due to the diversity of the tumours, it was not possible to establish a relationship between metalloproteinase-2 expression and tumour progression.


Full Text:

PDF

References


Aranha P.C., Hansen H.C., Rasmussen L.H., Strobel B.W. & Friis C. 2014. Determination of ptaquiloside and pterosin B derived from bracken (Pteridium aquilinum) in cattle plasma, urine and milk. Journal of Chromatography B. 1(951-952): 44-51.

Barros S.S.L.V. 2006. Expressão Imuno-histoquímica de metaloproteinases em carcinoma epidermóide de lábio inferior e língua. 128f. Natal, RN. Tese (Doutorado em Patologia Oral) - Programa de Pós-graduação em Patologia Oral, Universidade Federal do Rio Grande do Norte.

Benoit T., Keller E.X., Wolfsgruber P., Hermanns T., Günthart M., Banzola I., Sulser T., Provenzano M. & Poyet C. 2015. High VEGF-D and Low MMP-2 Serum Levels Predict Nodal-Positive Disease in Invasive Bladder Cancer. Medical Science Monitor. 21: 2266-2274.

Bergers G., Brekken R., McMahon G., Vu T.H., Itoh T., Tamaki K., Tanzawa K., Thorpe P., Itohara S., Werb Z. & Hanahan D. 2000. Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nature Cell Biology. 2(10): 737-744.

Birkedal-Hansen H., Moore W.G., Bodden M.K., Windsor L.J., Birkedal-Hansen B., DeCarlo A. & Engler J.A. 1993. Matrix Metalloproteinases: a Review. Critical Reviews in Oral Biology & Medicine. 4(2): 197-250.

Campo E., Merino M.J., Liotta L., Neumann R. & Stetler-Stevenson W. 1992. Distribution of the 72-kd type IV collagenase in nonneoplastic and neoplastic thyroid tissue. Human pathology. 23(12): 1395-1401.

Cavalheiro B.G. 2006. Expressão das metaloproteinases MMP-2, MT1-MMP e TIMP-2 e aspectos clinicopatológicos no carcinoma medular da glândula tireóide: implicações prognósticas. 248f. São Paulo, SP. Tese (Doutorado em Ciências) - Faculdade de Medicina, Universidade de São Paulo.

Chan C.C., Menges M., Orzechowski H.D., Orendain N., Pistorius G., Feifel G., Zeitz M. & Stallmach A. 2001. Increased matrix metalloproteinase 2 concentration and transcript expression in advanced colorectal carcinomas. International Journal of Colorectal Disease. 16(3): 133-140.

Clark I.M., Swingler T.E., Sampieri C.L. & Edwards D.R. 2008. The regulation of matrix metalloproteinases and their inhibitors. International Journal of Biochemistry & Cell Biology. 40(6-7): 1362-1378.

Clutterbuck A.L., Harris P., Allaway D. & Mobasheri A. 2001. Matrix metalloproteinases in inflammatory pathologies of the horse. Veterinary Journal. 183(1): 27-38.

Costa R.M.G., Bastos M.M., Oliveira P.A. & Lopes C. 2012. Bracken-associated human and animal health hazards: chemical, biological and pathological evidence. Journal of Hazardous Materials. 15: 203-204.

Davidson B., Goldberg I., Kopolovic J., Lerner-Geva L., Gotlieb W.H., Ben-Baruch G. & Reich R. 1999. MMP-2 and TIMP-2 expression correlates with poor prognosis in cervical carcinoma--a clinicopathologic study using immunohistochemistry and mRNA in situ hybridization. Gynecologic Oncology. 73(3): 372-382.

Drury R.A.B., Wallington E.A. & Cameron R. 1967. Connective Tissue Fibers. In: Carleton H.M., Drury R.A.B. & Wallington E.A. (Eds). Carleton’s Histological Techniques. 4th edn. New York: Oxford University Press, pp.166-181.

Ellis L.M. & Fidler I.J. 1996. Angiogenesis and metastasis. European Journal of Cancer. 32(14): 2451-2460.

Falbo M.K., Kommers G.D., Masuda E.K., Fighera R.A., Piazer J.V.M., Barros C.S.L., Martins T.B. & Rosa F.B. 2005. Alterações hematológicas, bioquímicas, urinárias e histopatológicas na intoxicação natural em bovinos pela samambaia Pteridium aquilnum (L.) Kühn. Semina. 26(7): 547-558.

Franchi A., Santucci M., Masini E., Sardi I., Paglierani M. & Gallo O. 2002. Expression of matrix metalloproteinase 1, matrix metalloproteinase 2, and matrix metalloproteinase 9 in carcinoma of the head and neck. Cancer. 95(9): 1902-1910.

Furlan F.H., Mendes E.R.S., Ducatti K.R., Marcon G.C., Dombrosky T., Amorim T.M. & Riet-Correa F. 2014. Intoxicação aguda por Pteridium aracnoideum e Pteridium caudatum em bovinos e distribuição das plantas em Mato Grosso. Pesquisa Veterinária Brasileira. 34(4): 343-348.

Galvão A., Brito M.F., Aragão A.P., Yamasaki E.M., Peixoto P.V. & Tokarnia C.H. 2012. Sobrevivência/viabilidade de bovinos com hematúria enzoótica após transferência para região livre de Pteridium. Pesquisa Veterinária Brasileira. 32(9): 887-902.

Hoque M., Somvanshi R., Singh G.R. & Mogha I.V. 2002. Ultrasonographic evaluation of urinary bladder in normal, fern fed and enzootic bovine haematuria-affected cattle. Journal of Veterinary Medicine. 49(8): 403-407.

Inoue Y., Ito K., Tereda T., Nishimura N., Hatazoe T. & Sato K. 2000. Degenerative changes in the endometrial vasculature of the mare detected by videoendoscopic examination. Proceedings of the AAEP Convention. 46: 325-329.

Kumar B., Koul S., Petersen J., Khandrika L., Hwa J.S., Meacham R.B., Wilson S. & Koul H.K. 2010. p38 mitogen-activated protein kinase–driven MAPKAPK2 regulates invasion of bladder cancer by modulation of MMP-2 and MMP-9 activity. Cancer Research. 70(2): 832-841.

Maiolino P., Ozkul A., Sepici-Dincel A., Roperto F., Yücel G., Russo V., Urraro C., Lucà R., Riccardi M.G., Martano M., Borzacchiello G., Esposito I. & Roperto S. 2013. Bovine papillomavirus type 2 infection and microscopic patterns of urothelial tumors of the urinary bladder in water buffaloes. BioMed Research International. 2013: 937918. DOI: 10.1155/2013/937918

Mccawley L. & Matrisian L.M. 2000. Matrix metalloproteinases: multifunctional contributors to tumor progression. Molecular Medicine Today. 6(4): 149-156.

Meuten D.J & Meuten T.L.K. 2020. Tumors of the Urinary system. In: Meuten D.J. (Ed). Tumors in domestic animals. 5th edn. Ames: Wiley-Blackwell, pp.632-688.

Morris J. & Dobson J. 2001. Urinary tract. In: Small Animal Oncology. Oxford: Blackwell Science, pp. 154-165.

Newman S.J., Confer A.W. & Panciera R.J. 2009. Sistema urinário. In: McGavin M.D. & Zachary J.F. (Eds). Bases de Patologia em Veterinária. 4.ed. Rio de Janeiro: Elsevier, pp.613-691.

Noronha L., Hecke F.A.F., Magalhães T.A., Pinheiro D.L. & Cat I. 2004. Estudo de 11 casos de fibroelastose endocárdica primária. Jornal Brasileiro de Patologia e Medicina Laboratorial. 40(4): 261-264.

Oliveira L.G.P. 2009. Novos aspectos patológicos e patogenéticos da hematúria enzoótica bovina. 141f. Seropédica, RJ. Dissertação (Mestrado em Clínica e Cirurgia Veterinária) - Programa de Pós-graduação em Medicina Veterinária, Universidade Federal Rural do Rio de Janeiro.

Papparella S., Restucci B., Paciello O. & Maiolino P. 2002. Expression of matrix metalloprotease-2 (MMP-2) and the activator membrane type 1 (MT1-MMP) in canine mammary carcinomas. Journal of Comparative Pathology. 126(4): 271-276.

Peixoto P.V., França T.N., Barros C.S.L. & Tokarnia C.H. 2003. Histopathological aspects of bovine enzootic hematuria in Brazil. Pesquisa Veterinária Brasileira. 23(2): 65-81.

Constable P.D., Hinchcliff K.W., Done S.H. & Grünberg W. 2017. Diseases of the Urinary System. In: Veterinary medicine: A textbook of the diseases of cattle, horses, sheep, pigs and goats, tenth. London: Saunders, pp.1095-1154.

Reis S.D.S., Macêdo J.T.S.A., Oliveira R.S., Peixoto T.C., Ferreira M.M., Gasper A.L. & Pedroso P.M.O. 2016. Enzootic hematuria in cattle from Northeastern Brazil. Brazilian Journal of Veterinary Pathology. 9(2): 78-82.

Ribeiro R.I.M.A, Borges Jr. P.C., Cardoso S.V., Candelori I., Espíndola F.S., Cassali G.D. & Loyola A.M. 2008. Expressão de metaloproteinases de matriz e de seus inibidores teciduais em carcinomas basocelulares. Jornal Brasileiro de Patologia e Medicina Laboratorial. 44(2): 115-121.

Roperto S., Brun R., Paolini F., Urraro C., Russo V., Borzacchiello G., Pagnini U., Raso C., Rizzo C., Roperto F. & Venuti A. 2008. Detection of bovine papillomavirus type 2 in the peripheral blood of cattle with urinary bladder tumours: possible biological role. Journal of General Virology. 89(12): 3027-3033.

Schmalfeldt B., Prechtel D., Härting K., Späthe K., Rutke S., Konik E., Fridman R., Berger U., Schmitt M., Kuhn W. & Lengyel E. 2001. Increased expression of matrix metalloproteinases (MMP)-2, (MMP)-9, and the urokinase-type plasminogen activator is associated with progression from benign to advanced ovarian cancer. Clinical Cancer Research. 7(8): 2394-2404.

Shigeru K., Miyata Y. & Kanetake H. 2006. Current status and perspective of antiangiogenic therapy for cancer: urinary cancer. International Journal of Clinical Oncology. 11(2): 90-107.

Silva D.A., Watanabe A.H.Q. & Marçal W.S. 2016. Avaliação epidemiológica da prevalência da hematuria enzoótica bovina associada a samambaia no Paraná. Publicações em Medicina Veterinária e Zootecnia. 10(2): 125-131.

Silva M.A. 2012. Caracterização histopatológica e imunoistoquímica de bexigas de bovinos com hematúria enzoótica. 110f. Alegre, ES. Dissertação (Mestrado em Ciências Veterinárias) - Programa de Pós-Graduação em Ciências Veterinária, Universidade Federal do Espírito Santo.

Silva M.A., Sequeira J.L., Madureira A.P. & Nunes L.C. 2013. Caracterização imunoistoquímica de neoplasias de bexigas associadas à hematúria enzoótica bovina. Semina. 34(1): 281-291.

Somvanshi R., Pathania S., Nagarajan N., Pangty K. & Kumar P. 2012. Pathological study of non-neoplastic urinary bladder lesions in cattle and buffaloes: a preliminary report. Tropical Animal Health and Production. 44(4): 855-861.

Souto M.A.M., Kommers G.D., Barros C.S.L., Rech R.R. & Piazer J.V.M. 2006. Neoplasmas da bexiga associados à hematúria enzoótica bovina. Ciência Rural. 36(5): 1647-1650.

Turpeenniemi-Hujanen T. 2005. Gelatinases (MMP-2 and-9) and their natural inhibitors as prognostic indicators in solid cancers. Biochimie. 87(3-4): 287-297.

Verhöeff F.H. 1908. Some new staining methods of wide applicability. Including a rapid differential stain for elastic tissue. Journal of American Medical Association. 11: 876-877.

Weidner N. & Folkman J. 1996. Tumoral vascularity as a prognostic factor in cancer. Important Advanced in Oncology. 1996: 167-190.

Westermarck J. & Kähäri V.M. 1999. Regulation of matrix metalloproteinase expression in tumor invasion. FASEB Journal. 13(8): 781-792.

Wosiacki S.R., Claus M.P., Alfieri A.F. & Alfieri A.A. 2006. Bovine papillomavius type 2 detection in the urinary bladder of cattle with chronic enzootic haematuria. Memórias do Instituto Oswaldo Cruz. 101(6): 635-638.

Yoshizaki T., Sato H. & Furukawa M. 2002. Recent advances in the regulation of matrix metalloproteinase 2 activation: from basic research to clinical implication. Oncology Reports. 9(3): 607-611.

Yoshizaki T., Maruyama Y., Sato H. & Furukawa M. 2001. Expression of tissue inhibitor of matrix metalloproteinase-2 correlates with activation of matrix metalloproteinase-2 and predicts poor prognosis in tongue squamous cell carcinoma. International Journal of Cancer. 95(1): 44-50.




DOI: https://doi.org/10.22456/1679-9216.100451

Copyright (c) 2020 Anderson Barros Archanjo, Natalia Viana Tamiasso, Juliana de Castro Cosme, Maria Aparecida da Silva, Leonardo Oliveira Trivilin, Julio Lopes Sequeira, Louisiane de Carvalho Nunes

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.