Evaluating the effects of Atorvastatin on cultured human endometrium in a three-dimensional fibrin matrix

Montaseri, Azadeh; Khazaei, Mozaffar; Ghorbani, Rostam; Rezaei, Mansour

Evaluating the effects of Atorvastatin on cultured human endometrium in a three-dimensional fibrin matrix

Vol. 7, Issue 4, / January-March 2007
(pages 358-366)

Azadeh Montaseri: - Reproduction Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Mozaffar Khazaei: - Reproduction Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Rostam Ghorbani: - Reproduction Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Mansour Rezaei: - Reproduction Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran

Received: 1/1/2007 Accepted: 1/1/2007 - Publisher : Avicenna Research Institute

Other Format

Abstract

Introduction: Endometriosis, as one of the main causes of infertility in women, has devoted a lot of research to itself. Three-dimensional culture of human endometrial tissue has been known as a functional model for endometriosis. Statins have a variety of different effects on angiogenesis and they have been suggested as a candidate for the inhibition of this disease. Therefore, the aim of the present study is to determine inhibitory or promotory effects of different concentrations of Atorvastatin on the growth of cultured human endometrium. Materials & Methods: 8 endometrial samples were cut into 1x1 mm pieces and were cultured in 24-well culture plates. To establish the desired three-dimensional model, 0.5 ml of fibrinogen solution in M199 medium, with a concentration of 3mg/ml and 10µL of thrombin were added to each well. After formation of the jelly, the endometrial fragments were placed on it and a second jelly layer was added. The endometrial tissue of each specimen was cultured in four 5-piece layers. The first row was regarded as control and to the second, third and forth rows, 0.1, 1.0 and 10µM concentrations of the medication were added respectively. Every 3 days, endometrial pieces and their probable changes were observed by an invert microscope (Motic-AE31) and the culture media were changed. The tissue fragments were also photographed on days 1, 7, 14 and 21. At the end of the study, the cultures were graded according to their cellular outgrowths, invasion of deposited cells into the jelly and growth of endometrial glands and images related to the 3-D cultures were analyzed. To confirm the growth of cellular structures into the jelly, paraffin embedded cuts of the cultured specimen were immunohistochemically stained by anti-vimentin, anticytokeratin and anti-CD31. The data were analyzed by SPSS statistical software (Version 11.5). The statistical analysis related to the grading of wells was done by Freedman test and the statistical analysis of the images was performed by one-way ANOVA. Results: Growth changes were 82.5% in the control group and 87.5% in the 0.1µM concentration group, depicting significantly higher values compared to the control group (p<0.05 ). the changes in 1µm and in 10µm groups were 50% and 5% respectively which had significant differences with that of the control group (p<0.001). conclusion: different concentrations of atorvastatin have different effects on the growth of cultured endometrial tissue, 0.1µm concentration increases growth of cultured human endomet-rium, while 1µm and 10µm can decrease growth of the tissue. further animal studies on this subject are suggested.< pan>

Keywords: Atorvastatin, Endometrium, Endometriosis, 3D-culture, Tissue growth, Fibrin, Thrombin

To cite this article:

Montaseri A, Khazaei M, Ghorbani R, Rezaei M. Evaluating the effects of Atorvastatin on cultured human endometrium in a three-dimensional fibrin matrix. J Reprod Infertil. 2007;7(4):358-366.

References

Lebovic DI, Bentzien F, Chao VA, Garrett ET, Taylor RN. Induction of an angiogenic phenotype in endomet-riotic stromal cell culture by interlukine-1 beta. Mol Hum Reprod. 2000;6:269-75. [PubMed]
Strathy JH, Mdgaard CA, Coulam CB, Melton LJ. Endometriosis and infertility: A laparoscopic study of endometriosis among fertile and infertile women. Fertil Steril. 1982;38:667-72. [PubMed]
Kettle LM, Hummel WP. Modern medical management of endometriosis. Obset Gynecol Clin North Am. 1997; 24:361-73. [PubMed]
Adamson GD, Nelson HP. Surgical treatment of endo-metriosis. Obset Gynecol Clin North Am. 1997;24: 375-409. [PubMed]
Sampson JA. Peritoneal endometriosis due to menstrual dissemination of endometrial tissue into peritoneal cavity. Am J Obset Gynecol. 1927;14:422-9.
Healy DL, Rogers PAW, Hill L, Wingfield M. Angio-genesis: a new theory for endometriosis. Hum Reprod Update. 1998;4(5):736-40. [PubMed]
Fasciani A, Bocci G, Xu J, Bielecki R, Greenblatt E, Leyland N, Casper R C. Three-dimensional in vitro culture of endometrial explants mimics the early stage of endometriosis. Fertil Steril. 2003;80(5):1137-43. [PubMed]
Khazaei M, Esfandiari N, Javed M, Gotlieb L, Casper RC. Successful of human secretory phase endometrium culture as an In vitro model for endometriosis. Fertil Steril. 2004;82(2):S164.
Khazaei M, Esfandiari N, Gotlieb L, Casper RC. Angiogenesis following three-dimensional culture of isolated human endometrial stroma cells. Fertil Steril. 2004; 82 (2): S61.
Lea AP, Mc Tavish D. Atorvastatin. A review of its pharmacology and therapeutic potential in the manage-ment of hyperlipidemias. Drugs. 1997;35(5):828-47. [PubMed]
Graaf MR. The risk of cancer in users of statins. J Clin Oncol. 2004;22(12):2388-94. [PubMed]
Ruiz-Velasco. Statins upregulate CD 36 expressin in human monocytes an affect strengthened when com-bined with PPAR-gamma ligand putative contribution of Rho GTPase in statin-induced CD36 expression. Biochem Pharmacol. 2004;67(2):303-13.
Laufs U. Beyond lipid-lowering: effects of statins on endothelial nitric oxide. Eur J Clin Pharmacol. 2003; 58(11): L719-31. [PubMed]
Esfandiari N, Khazaei M, Ai j, Bielecki R, Gotlieb L, Ryan E, Casper R. Effect of statin on an in vitro model of endometriosis. Fertil Steril. 2006;87(2):257-62. [PubMed]
Wierzbicki AS. Atorvastatin. Expert Opin Pharmaco-ther. 2001;2(5):819-30. [PubMed]
Malinowski JM.: Atorvastatin: a hydroxylmethylgl-utaryl-coenzyme A reductase inhibitpor. Am J Health Syst Pharm. 1998;55(21):2253-67. [PubMed]
Vincent L, Chen W, Hong L. Inhibition of endothelial cell migration by cerivastatin, an HMG-CoA reductase inhibitor: contribution to its anti-angiogenic effect. FEBS Lett. 2001;495:159-66. [PubMed]
Kureishi Y, Luo Z, Shiojima I. The HMG-CoA reduc-tase inhibitor simvastatin activates the protein kinase akt and promotes angiogenesis in normocholeste-rolemic animals. Nat Med. 2000;6:1004-10. [PubMed]
Weis M, Heeschen C, Glassford A, Cooke J. Statins have Biphasic effects on angiogenesis. Circulation. 2002;105:739-46. [PubMed]
Rorowski A, Walsh K. Statin therapy and angiogene-sis. Curr Opin Lipidol. 2003;14(6):599-603. [PubMed]
Walter DH, Zeiher AM, Dimmeler S. Effects of statins on endothelium and their contribution to neovasculariza-tion by mobilization of endothelial progenitor cells. Coron Artery Dis. 2004;15(5):235-242. [PubMed]
Littman BA, Smotrich DB, Stillman RJ. Endometrio-sis; Principal and practice of endocrinology and metabolism, 2nd Edn. Philadelphia JB. Lippincott Com-pany. 1995;906-9.