Sıçanlarda oluşturulan cerrahi müdahalesiz yeni bir deneysel Asherman sendromu modeli

Öz

Amaç: Asherman sendromu(AS) uterin kaviteye travma sonucu kavitenin adezyonlarla kısmi ya da tam tıkanması durumudur. Klinik öncesi çalışmalarla yeni tedavi yöntemlerinin etkinliğini gösterebilmek için öncelikle AS modelinin hayvanda oluşturulmasına ihtiyaç vardır. Bu çalışmanın amacı, AS ve intrauterin adezyon modellemesinde kullanılmak üzere cerrahi uygulanmaksızın yeni ve etkili bir yöntem geliştirmek, bu şekilde araştırmacılara güvenlik ve uygulanabilirlik açısından daha etkin bir metot önerebilmektir.

Yöntem: On iki dişi Wistar Albino sıçan rastgele 2 gruba ayrıldı. Daha önceden hazırlanmış olan pembe renk (20 gauge) kateterle transvajinal yoldan hayvanların sol uterin hornularına ulaşıldı. Grup I’deki hayvanlara 0,2 ml serum fizyolojik uygulandı, Grup II’dekilere ise 0,2 ml Triklorasetik asit (TCA) uygulandı. Üç menstruel siklus beklendikten sonra hayvanların uterusları çıkarılıp Hematoksilen-Eozin ve Masson Trikrom boyamalar yapılarak histopatolojik olarak değerlendirildi. Yapılan Hematoksilen-Eozin boyalı kesitlerde inflamasyon dereceleri değerlendirilirken, Masson Trikrom boyalı kesitlerde fibrozis dereceleri değerlendirildi.

Bulgular: Grup 1’e ait uterus kesitleri normal histolojik görünüme sahipken, Grup 2’ye ait sol uterus kesitlerinde inflamasyon ve fibrozis oluştuğu gözlendi. Bu sonuçlar istatistiksel olarak anlamlıydı (p=0,002).

Sonuç: Sonuç olarak, önerdiğimiz cerrahi müdahalesiz AS modelleme yöntemi, literatürde önerilen yöntemler ile benzer şekilde hastalık oluşturmuş ve bu etki, yapılan histopatolojik incelemelerle de ortaya konmuştur. Bu şekilde, AS için doğru bir hastalık modellemesi gerçekleştiren cerrahisiz yeni bir yöntem önerilmiştir.

Anahtar Kelimeler

• Asherman sendromu,sıçan,jinatrezi,patoloji,hayvan modeli

A new nonsurgical experimental model for Asherman syndrome created in rats

Abstract

Aim: Asherman Syndrome (AS) is a partial or complete obstruction of the uterine cavity with adhesions as a result of trauma. In pre-clinical studies, to be able to show the effectiveness of new treatment methods, first of all, the AS model needs to be created in the animals. The aim of this study is to develop a new and effective nonsurgical method for using in AS and intrauterine adhesions modeling, and through this way, to propose a more effective method for researchers in terms of safety and feasibility.

Methods: Twelve female Wistar Albino rats were divided into two groups. It was reached to the left uterine horn transvaginally by using pre-prepared pink color (20 gauge) cannula. While 0.2 ml normal saline was applied to the animals in Group I (control group), 0.2 ml (Trichloroacetic acid) TCA was applied to the animals in Group II (experiment group).The right uterine horns of the animals were left without treatment. After three menstrual cycles, the animals were sacrificed and Hematoxylin-Eosin and Masson’s Trichrom staining were performed and evaluated histopathologically. Inflammation was evaluated in Hematoxylin-Eosin staining and fibrosis was evaluated in Masson’s Trichrom staining.

Results: Whereas the uterine sections of the Group I have normal histologic appearance, inflammation and fibrosis were found in the left uterine sections of the Group 2 by histopathological evaluation. Results were statistically significant (p=0.002).

Conclusion: The proposed nonsurgical AS modeling method created disease, and this was also revealed by histopathological examinations. Through this way, a new AS model is suggested without surgery, in which the disease is correctly created.

Keywords

• Asherman Syndrome,rat,gynatresia,pathology,animal model

References

1. 1. Lin X, Wei M, Li TC, Huang Q, Huang D, Zhou F, et al. A comparison of intrauterine balloon, intrauterine contraceptive device and hyaluronic acid gel in the prevention of adhesion reformation following hysteroscopic surgery for Asherman syndrome: a cohort study. Eur J Obstet Gynecol Reprod Biol. 2013;170:512-6.
2. 2. Yu D, Wong YM, Cheong Y, Xia E, Li TC. Asherman syndrome—one century later. Fertil Steril. 2008;89:759-79.
3. 3. March CM. Management of Asherman’s syndrome. Reprod Biomed Online. 2011;23:63-76.
4. 4. Al-Inany H. Intrauterine adhesions. An update. Acta Obstet Gynecol Scand. 2001;80:986-93.
5. 5. Myers EM, Hurst BS. Comprehensive management of severe Asherman syndrome and amenorrhea. Fertil Steril. 2012; 97:160-4.
6. 6. Yu D, Li TC, Xia E, Huang X, Liu Y, Peng X. Factors affecting reproductive outcome of hysteroscopic adhesiolysis for Asherman's syndrome. Fertil Steril. 2008;89:715-22.
7. 7. Broome JD, Vancaillie TG. Fluoroscopically guided hysteroscopic division of adhesions in severe Asherman syndrome. Obstet Gynecol. 1999;93:1041-3.
8. 8. Tsui KH, Lin LT, Cheng JT, Teng SW, Wang PH. Comprehensive treatment for infertile women with severe Asherman syndrome. Taiwan J Obstet Gynecol. 2014;53:372-5.

9. 9. Tu CH, Yang XL, Qin XY, Cai LP, Zhang P. Management of intrauterine adhesions: a novel intrauterine device. Med Hypotheses. 2013;81:394-6.
10. 10. Kuramoto G, Takagi S, Ishitani K, Shimizu T, Okano T, Matsui H. Preventive effect of oral mucosal epithelial cell sheets on intrauterine adhesions. Hum Reprod. 2014;30:406-16.
11. 11. Wang J, Ju B, Pan C, Gu Y, Zhang Y, Sun L, et al. Application of bone marrow-derived mesenchymal stem cells in the treatment of intrauterine adhesions in rats. Cell Physiol Biochem. 2016;39:1553-60.
12. 12. Tan J, Li P, Wang Q, Li Y, Li X, Zhao D, et al. Autologous menstrual blood-derived stromal cells transplantation for severe Asherman's syndrome. Hum Reprod. 2016;31:2723-9.
13. 13. Jing Z, Qiong Z, Yonggang W, Yanping L. Rat bone marrow mesenchymal stem cells improve regeneration of thin endometrium in rat. Fertil Steril. 2014;101:587-94.
14. 14. Kilic S, Yuksel B, Pinarli F, Albayrak A, Boztok B, Delibasi T. Effect of stem cell application on Asherman syndrome, an experimental rat model. J Assist Reprod Genet. 2014;31:975-82.
15. 15. Zhao J, Zhang Q, Wang Y, Li Y. Uterine infusion with bone marrow mesenchymal stem cells improves endometrium thickness in a rat model of thin endometrium. Reprod Sci. 2015;22:181-8.
16. 16. Alawadhi F, Du H, Cakmak H, Taylor HS. Bone Marrow-Derived Stem Cell (BMDSC) transplantation improves fertility in a murine model of Asherman's syndrome. PloS One. 2014;9:e96662.
17. 17. Yüksel B, Kılıç S, Boztok B, Albayrak A. Deneysel Asherman Sendromu, Rat Modeli. Turkiye Klinikleri J Gynecol Obst. 2014;24:195-7.
18. 18. Cervelló I, Gil-Sanchis C, Santamaría X, Cabanillas S, Diaz A, Faus A, et al. Human CD133+ bone marrow-derived stem cells promote endometrial proliferation in a murine model of Asherman syndrome. Fertil Steril. 2015;104:1552-60.
19. 19. Liu Y, Tal R, Pluchino N, Mamillapalli R, Taylor HS. Systemic administration of bone marrow-derived cells leads to better uterine engraftment than use of uterine-derived cells or local injection. J Cell Mol Med. 2018;22:67-76.
20. 20. Xu Q, Duan H, Gan L, Liu X, Chen F, Shen X, et al. MicroRNA-1291 promotes endometrial fibrosis by regulating the ArhGAP29-RhoA/ROCK1 signaling pathway in a murine model. Mol Med Rep. 2017;16:4501-10.
21. 21. Bazoobandi S, Tanideh N, Rahmanifar F, Tamadon A, Keshtkar M, Mehrabani D, et al. Induction of Asherman's syndrome in rabbit. J Reprod Infertile. 2016;17:10.
22. 22. Jing Z, Hong G, Yanping L. Development of an animal model for thin endometrium using 95% ethanol. J Fert In Vitro. 2012;2:4.
23. 23. Tang YQ, Gan L, Xu Q, Wang S, Li JJ, Duan H. Effects of human umbilical cord mesenchymal stem cells on intrauterine adhesions in a rat model. Int J Clin Exp Pathol. 2016;9:12119-29.
24. 24. Wang X, Ma N, Sun Q, Huang C, Liu Y, Luo X. Elevated NF-κB signaling in Asherman syndrome patients and animal models. Oncotarget. 2017;8:15399.
25. 25. Liu F, Zhu ZJ, Li P, He YL.Creation of a female rabbit model for intrauterine adhesions using mechanical and infectious injury. J Surg Res. 2013;183:296-303.
26. 26. Khrouf M, Morel O, Hafiz A, Chavatte-Palmer P, Fernandez H. Evaluation of the rabbit as an experimental model for human uterine synechia. J Hum Reprod Sci. 2012;5:175.
27. 27. Kaya C, Sever N, Cengiz H, Yildiz S, Ekin M, Yasar L. A randomized controlled study of the efficacy of misoprostol and hyaluronic acid in preventing adhesion formation after gynecological surgery: a rat uterine horn model. Eur J Obstet Gynecol Reprod Biol. 2014;176:44-9.
28. 28. Judd MD, Hill PD, Potter LA, Bown SG, McColl I. Destruction of the rabbit endometrium using a low-powered Neodymium-YAG laser. Lasers Med Sci. 1991;6:133-40.
29. 29. Jang HY, Myoung SM, Choe JM, Kim T, Cheon Y, Kim YM, et al. Effects of Autologous Platelet-Rich Plasma on Regeneration of Damaged Endometrium in Female Rats. Yonsei Med J. 2017;58:1195-203.
30. 30. Mescher AL. Junqueira’s Basic Histology. 13th Ed. New York: Mc Graw Hill Education; 2013. p.463.
31. 31. Schenker JG. Etiology of and therapeutic approach to synechia uteri. Eur J Obstet Gynecol Reprod Biol. 1996;65:109–13.
32. 32. Roy KK, Baruah J, Sharma JB, Kumar S, Kachawa G, Singh N. Reproductive outcome following hysteroscopic adhesiolysis in patients with infertility due to Asherman’s syndrome. Arch Gynecol Obstet. 2010;281:355–61.
33. 33. Fernandez H, Peyrelevade S, Legendre G, Faivre E, Deffieux X, Nazac A. Total adhesions treated by hysteroscopy: must we stop at two procedures. Fertil Steril. 2012;98:980–5.