Reliability of histopathological examination and immunohistochemistry of a single biopsy for evaluation of endometrial health in Icelandic mares
Abstract
Introduction. Endometritis and endometrosis have been and still are the major reasons for infertility in mares. The diagnosis of endometritis can be based on cytology and microbiology, but endometrial biopsy is still the only way to diagnose endometrosis in the mare. Our study attempted to determine if a single biopsy using histopathology and immunohistochemistry is sufficient to ascertain reasons for infertility in Icelandic mares. The objectives of this study were to examine the relationship between deviations in endometrial biopsies in terms of prostaglandin-endoperoxide synthase-2 (PTGS-2) and fibronectin expression and polymorphonuclear cells (PMNs) infiltration, as well as scoring degeneration in two endometrial biopsies.
Material and methods. Materials were collected from 53 Icelandic breed mares, from whom two endometrial biopsies were collected and they were used for histopathology and for immunohistochemistry for PTGS-2 and fibronectin.
Results. In our study, twenty-six of 53 mares (49%) showed differences in the biopsy score between the left and the right uterine horns (p = 0.002). There were statistically significant differences in fibronectin expression (p = 0.001), as well as in PTGS-2 expression in the superficial epithelium (p = 0.017).
Conclusions. Significant differences in the biopsy score, and fibronectin and PTGS-2 expression, between two endometrial biopsies obtained from individual mares demonstrated that a single biopsy could be insufficient for diagnosing uterine health status in Icelandic mares.
Keywords: mareinfertilityendometritishistopathologyfibronectinprostaglandin-endoperoxide synthase- 2IHC
References
- Walter I, Handler J, Reifinger M, et al. Association of endometriosis in horses with differentiation of periglandular myofibroblasts and changes of extracellular matrix proteins. Reproduction. 2001; 121(4): 581–586.
- Liu IKM, Troedsson MHT. The diagnosis and treatment of endometritis in the mare: yesterday and today. Theriogenology. 2008; 70(3): 415–420.
- Traub-Dargatz JL, Salman MD, Voss JL. Medical problems of adult horses, as ranked by equine practitioners. J Am Vet Med Assoc. 1991; 198(10): 1745–1747.
- Schlafer DH. Equine endometrial biopsy: enhancement of clinical value by more extensive histopathology and application of new diagnostic techniques? Theriogenology. 2007; 68(3): 413–422.
- Schoon HA, Schoon D, Klug E. The endometrial biopsy in the mare with regard to clinical correlations. Pferdeheilkunde Equine Medicine. 1997; 13(5): 453–464.
- LeBlanc MM, Causey RC. Clinical and subclinical endometritis in the mare: both threats to fertility. Reprod Domest Anim. 2009; 44(Suppl 3): 10–22.
- Ricketts SW, Barrelet A. A retrospective review of the histopathological features seen in a series of 4241 endometrial biopsy samples collected from UK Thoroughbred mares over a 25 yearperiod. Pferdeheilkunde Equine Medicine. 1997; 13(5): 525–530.
- Snider TA, Sepoy C, Holyoak GR. Equine endometrial biopsy reviewed: Observation, interpretation, and application of histopathologic data. Theriogenology. 2011; 75(9): 1567–1581.
- Buczkowska J, Kozdrowski R, Nowak M, et al. Comparison of the biopsy and cytobrush techniques for diagnosis of subclinical endometritis in mares. Reprod Biol Endocrinol. 2014; 12: 27.
- Kozdrowski R, Sikora M, Buczkowska J, et al. Effects of cycle stage and sampling procedure on interpretation of endometrial cytology in mares. Anim Reprod Sci. 2015; 154: 56–62.
- Nielsen JM. Endometritis in the mare: a diagnostic study comparing cultures from swab and biopsy. Theriogenology. 2005; 64(3): 510–518.
- Kenney RM, Doig PA. Equine endometrial biopsy. In: Marrow D. ed. Current therapy in theriogenology. WB Sounders, Philadelphia 1986: 723–729.
- Walter I, Handler J, Miller I, et al. Matrix metalloproteinase 2 (MMP-2) and tissue transglutaminase (TG 2) are expressed in periglandular fibrosis in horse mares with endometrosis. Histol Histopathol. 2005; 20(4): 1105–1113.
- Hoffmann C, Ellenberger C, Mattos R, et al. The equine endometrosis: New insights into the pathogenesis. Animal Reproduction Science. 2009; 111(2-4): 261–278.
- Aresu L, Benali S, Giannuzzi D, et al. The role of inflammation and matrix metalloproteinases in equine endometriosis. J Vet Sci. 2012; 13(2): 171.
- Krzyzanowska- Golab D, Lemańska-Perek A and Kątnik-Prastowska I. Fibronektyna jako aktywny składnik macierzy pozakomórkowej. Postepy Hig Med Dosw. 2007; 61: 655–663.
- Iwahashi M, Muragaki Y, Ooshima A, et al. Alterations in distribution and composition of the extracellular matrix during decidualization of the human endometrium. J Reprod Fertil. 1996; 108(1): 147–155.
- Kaidi R, Brown P, David J, et al. Uterine collagen during involution in cattle. Matrix. 1991; 11(2): 101–107.
- Boos A. Immunohistochemical demonstration of collagen types I, III, IV, V and VI in the bovine endometrium during estrus cycle and after superovulation. Anatomia Histologia Embryologia. 1994; 23: 64.
- Boerboom D, Brown KA, Vaillancourt D, et al. Expression of key prostaglandin synthases in equine endometrium during late diestrus and early pregnancy. Biol Reprod. 2004; 70(2): 391–399.
- Ma W, Quirion R. Up-regulation of interleukin-6 induced by prostaglandin E from invading macrophages following nerve injury: an in vivo and in vitro study. J Neurochem. 2005; 93(3): 664–673.
- Fiala SM, Esmeraldino A, Jobim MIM, et al. Endometrial fibrotic changes. Is one biopsy enough to diagnose degenerative changes? Anim Reprod Sci. 2010; 121(Suppl): 89–90.
- Overbeck W, Jäger K, Schoon HA, et al. Comparison of cytological and histological examinations in different locations of the equine uterus-an in vitro study. Theriogenology. 2013; 79(9): 1262–1268.
- Sikora M, Król J, Nowak M, et al. The usefulness of uterine lavage and acute phase protein levels as a diagnostic tool for subclinical endometritis in Icelandic mares. Acta Vet Scand. 2016; 58(1): 50.
- Ricketts SW, Alonso S. Assessment of the breeding prognosis of mares using paired endometrial biopsy techniques. Equine Vet J. 1991; 23(3): 185–188.
- LeBlanc MM, Magsig J, Stromberg AJ. Use of a low-volume uterine flush for diagnosing endometritis in chronically infertile mares. Theriogenology. 2007; 68(3): 403–412.
- Christoffersen M, Brandis L, Samuelsson J, et al. Diagnostic double-guarded low-volume uterine lavage in mares. Theriogenology. 2015; 83(2): 222–227.
- Meuten DJ, Moore FM, George JW. Mitotic count and the field of view area: time to standardize. Vet Pathol. 2016; 53(1): 7–9.
- Palm F, Walter I, Budik S, et al. Influence of different semen extenders and seminal plasma on PMN migration and on expression of IL-1beta, IL-6, TNF-alpha and COX-2 mRNA in the equine endometrium. Theriogenology. 2008; 70(5): 843–851.
- Schoon HA, Schoon D. The Category I mare (Kenney and Doig 1986): Expected foaling rate 80-90% — fact or fiction? Pferdeheilkunde. 2003; 19: 698–701.
- Sappino AP, Schürch W, Gabbiani G. Differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab Invest. 1990; 63(2): 144–161.
- Schmitt-Gräff A, Desmoulière A, Gabbiani G. Heterogeneity of myofibroblast phenotypic features: an example of fibroblastic cell plasticity. Virchows Arch. 1994; 425(1): 3–24.
- Mansour GD, Henry M, Ferreira A. Immunohistochemical study of equine endometrial extracellular matrix during the oestrous cycle. J Comp Path. 2003; 129(4): 316–319.
- Arosh JA, Parent J, Chapdelaine P, et al. Expression of cyclooxygenases 1 and 2 and prostaglandin E synthase in bovine endometrial tissue during the estrous cycle. Biol Reprod. 2002; 67(1): 161–169.
- Atli M, Kurar E, Kayis S, et al. Evaluation of genes involved in prostaglandin action in equine endometrium during estrous cycle and early pregnancy. Animal Reprod Sci. 2010; 122(1-2): 124–132.
- Keith L, Ball BA, Scoggin K, et al. Diestrus administration of oxytocin prolongs luteal maintenance and reduces plasma PGFM concentrations and endometrial COX-2 expression in mares. Theriogenology. 2013; 79(4): 616–624.
- Troedsson M, Liu I, Crabo BG. Sperm transport and survival in the mare. Theriogenology. 1998; 49(5): 905–915.
- Madill S, Troedsson MH, Alexander SL, et al. Simultaneous recording of pituitary oxytocin secretion and myometrial activity in oestrous mares exposed to various breeding stimuli. J Reprod Fertil Suppl. 2000(56): 351–361.
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