Immunohistochemical study of matrix metalloproteinases 2 and 9 in the placenta of spontaneous miscarriage

Year 2025, Volume: 29 Issue: 5 , 2064 - 2076 , 01.09.2025

Russul Hassan , Mukhtar K. Haba

https://doi.org/10.12991/jrespharm.1766301

https://izlik.org/JA52DB39BU

Abstract

Miscarriage is the termination of pregnancy by spontaneous expulsion of embryo/fetus before 25 weeks of gestation or when the fetus weighs less than 500 grams. It is the most common pregnancy problem, impacting around 10% to 15% of all pregnancies. Matrix metalloproteinase 2 and 9 (MMP-2 and MMP-9) play a crucial role in embryo implantation and placentation; spontaneous abortion has been observed when there is abnormal expression of MMP-2 and MMP-9 levels. This research aims to evaluate the expression of MMP-2 and MMP-9 in the placenta of women with spontaneous miscarriage. The placental specimens were collected from 30 women who had delivered normally as a control group, 30 women who had miscarried in the first trimester, and 30 women who had miscarried in the second trimester. The specimens included the decidua basalis and trophoblast. The expression patterns of MMP-2 and MMP-9 were examined using the immunohistochemical study. Antibodies were used against MMP2 and MMP9. The results revealed a strong expression of MMP-2 and MMP-9 in the placenta of normally delivered women; 18 and 20 cases of first and second-trimester miscarriage showed weak expression of MMP-2, while 12 and 10 cases of both trimesters revealed moderate expression. MMP-9 showed weak expression in decidua and trophoblastic tissue of 26 and 24 cases of the first and second trimester, 4 and 6 specimens reported moderate expression in the first and second-trimester miscarriage, and significant differences were detected between the study groups (p-value <0.01). We conclude that MMP-2 and MMP-9 immunopositivity could indicate uncontrolled invasion of trophoblasts, leading to their destructive invasion and contributing to spontaneous miscarriage pathogenesis.

Keywords

Immunohistochemistry , Miscarriage , MMP2 , MMP9 , Placenta

References

• [1] Erra S, Merlo C. Early Miscarriage: Morphological Data Useful to Clinical and Nosographic Classification. Int J Appl Pharm Sci Res. 2019;4(2):13-16. https://doi.org/10.21477/ijapsr.4.2.1.
• [2] Linnakaari R, Helle N, Mentula M, Bloigu A, Gissler M, Heikinheimo O, Niinimäki M. Trends in the incidence, rate and treatment of miscarriage-nationwide register-study in Finland, 1998-2016. Hum Reprod. 2019;34(11):2120- 2128. https://doi.org/10.1093/humrep/dez211.
• [3] Woolner AM, Nagdeve P, Raja EA, Bhattacharya S, Bhattacharya S. Family history and risk of miscarriage: A systematic review and meta‐analysis of observational studies. Acta Obstet Gynecol Scand. 2020;99(12):1584-1594. https://doi.org/10.1111/aogs.13940.
• [4] Staun-Ram E, Goldman S, Gabarin D, Shalev E. Expression and importance of matrix metalloproteinase 2 and 9 (MMP-2 and-9) in human trophoblast invasion. Reprod Biol Endocrinol. 2004;2:59. https://doi.org/10.1186%2F1477- 7827-2-59.
• [5] Castruita-De la Rosa C, Garza-Veloz I, Delgado-Enciso I, Olivas-Chavez JC, Cardenas-Vargas E, Rodriguez-Sanchez IP, Francisco Citalan-Madrid A, Ortega-Cisneros V, Isaias Badillo-Almaraz J, Maria Trejo-Ortiz P, Araujo-Espino R, Araujo-Conejo A, de Jesus Jaime-Guzman J, Martinez-Fierro ML. Spontaneous abortion is preceded by an altered serum concentration of matrix metalloproteinases. J Matern Fetal Neonatal Med. 2020;33(24):4108-4116. https://doi.org/10.1080/14767058.2019.1597046.
• [6] Jing M, Chen X, Qiu H, He W, Zhou Y, Li D, Wang D, Jiao Y, Liu A. Insights into the immunomodulatory regulation of matrix metalloproteinase at the maternal-fetal interface during early pregnancy and pregnancy-related diseases. Front Immunol. 2023;13:1067661. https://doi.org/10.3389/fimmu.2022.1067661.
• [7] Zhang SM, Tian FJ, Zeng WH, Ma XL, Ren JB, Lin Y. XCL 1‐XCR 1 pathway promotes trophoblast invasion at maternal‐fetal interface by inducing MMP‐2/MMP‐9 activity. Am J Reprod Immunol. 2018;80(3):e12990. https://doi.org/10.1111/aji.12990.
• [8] Singh M, Kindelberger D, Nagymanyoki Z, Ng SW, Quick CM, Elias KM, Yamamoto H, Fichorova R, Fulop V, Berkowitz RS. Matrix metalloproteinases and their inhibitors and inducer in gestational trophoblastic diseases and normal placenta. Gynecol Oncol. 2011;122(1):178-182. https://doi.org/10.1016/j.ygyno.2011.03.025.
• [9] Nikolov A, Popovski N. Role of Gelatinases MMP-2 and MMP-9 in Healthy and Complicated Pregnancy and Their Future Potential as Preeclampsia Biomarkers. Diagnostics (Basel). 2021;11(3):480. https://doi.org/10.3390/diagnostics11030480.
• [10] Riley SC, Leask R, Chard T, Wathen NC, Calder AA, Howe DC. Secretion of matrix metalloproteinase-2, matrix metalloproteinase-9 and tissue inhibitor of metalloproteinases into the intrauterine compartments during early pregnancy. Mol Hum Reprod. 1999;5(4):376-381. https://doi.org/10.1093/molehr/5.4.376.
• [11] Zhang S, Mesalam A, Joo MD, Lee KL, Hwang JY, Xu L, Song SH, Koh PO, Yuan YG, Lv W, Kong IK. Matrix metalloproteinases improves trophoblast invasion and pregnancy potential in mice. Theriogenology. 2020;151:144- 150. https://doi.org/10.1016/j.theriogenology.2020.02.002.
• [12] Demir-Weusten AY, Seval Y, Kaufmann P, Demir R, Yucel G, Huppertz B. Matrix metalloproteinases-2,-3 and-9 in human term placenta. Acta histochem. 2007;109(5):403-412. https://doi.org/10.1016/j.acthis.2007.04.001
• [13] Staun-Ram E, Shalev E. Human trophoblast function during the implantation process. Reprod Biol Endocrinol. 2005;3:56. https://doi.org/10.1186/1477-7827-3-56.
• [14] Suryawan A, Mose JC, Handono B. MMP-9 level comparation between spontaneous abortion and normal pregnancy. Indones Biomed J. 2012;4(3):174-179. https://dx.doi.org/10.18585/inabj.v4i3.178
• [15] Nissi R, Talvensaari-Mattila A, Kotila V, Niinimäki M, Järvelä I, Turpeenniemi-Hujanen T. Circulating matrix metalloproteinase MMP-9 and MMP-2/TIMP-2 complex are associated with spontaneous early pregnancy failure. Reprod Bio Endocrinol. 2013;11:1-6. https://doi.org/10.1186/1477-7827-11-2.
• [16] Zhang Y, Li P, Guo Y, Liu X, Zhang Y. MMP-9 and TIMP-1 in placenta of hypertensive disorder complicating pregnancy. Exp Ther Med. 2019;18(1):637-641. https://doi.org/10.3892%2Fetm.2019.7591.
• [17] Timokhina E, Strizhakov A, Ibragimova S, Gitel E, Ignatko I, Belousova V, Zafiridi N. Matrix Metalloproteinases MMP-2 and MMP-9 Occupy a New Role in Severe Preeclampsia. J Pregnancy. 2020;2020:8369645. https://doi.org/10.1155/2020/8369645.
• [18] Behforouz A, Dastgheib SA, Abbasi H, Karimi-Zarchi M, Javaheri A, Hadadan A, Tabatabaei RS, Meibodi B, Neamatzadeh H. Association of MMP-2, MMP-3, and MMP-9 Polymorphisms with Susceptibility to Recurrent Pregnancy Loss. Fetal Pediatr Pathol. 2021;40(5):378-386. https://doi.org/10.1080/15513815.2019.1710879.
• [19] Karachrysafi S, Georgiou P, Kavvadas D, Papafotiou F, Isaakidou S, Grammatikakis IE, Papamitsou T. Immunohistochemical study of MMP-2, MMP-9, EGFR and IL-8 in decidual and trophoblastic specimens of recurrent pregnancy loss cases. J Matern Fetal Neonatal Med. 2023 ;36(1):2218523. https://doi.org/10.1080/14767058.2023.2218523.
• [20] Balci M, Özdemir G. Differential expression of EGFR-1, MMP-3, and MMP-9 in spontaneous abortions, induced abortions, and tubal pregnancies. Turk Patoloji Derg. 2019;35(1):1-8. https://doi.org/10.5146/tjpath.2018.01432.
• [21] Jiang G, Qi Y. Detection of MMP-9 and TIMP-3 mRNA expression in the villi of patients undergoing early spontaneous abortion: A report of 30 cases. Exp Ther Med. 2015;9(5):1939-1943. https://doi.org/10.3892/etm.2015.2336.
• [22] Staun-Ram E, Goldman S, Gabarin D, Shalev E. Expression and importance of matrix metalloproteinase 2 and 9 (MMP-2 and -9) in human trophoblast invasion. Reprod Biol Endocrinol. 2004 ;2:59. https://doi.org/10.1186%2F1477-7827-2-59.
• [23] Lin C, He H, Cui N, Ren Z, Zhu M, Khalil RA. Decreased uterine vascularization and uterine arterial expansive remodeling with reduced matrix metalloproteinase-2 and-9 in hypertensive pregnancy. Am J Physiol Heart Circ Physiol. 2020;318(1):H165-H180. https://doi.org/10.1152/ajpheart.00602.2019.
• [24] Shokry M, Omran OM, Hassan HI, Elsedfy GO, Hussein MRA. Expression of matrix metalloproteinases 2 and 9 in human trophoblasts of normal and preeclamptic placentas: preliminary findings. Exp Mol Pathol. 2009;87(3):219-225. https://doi.org/10.1016/j.yexmp.2009.08.001.
• [25] Aldafaay AAA, Abdulamir HA, Abdulhussain HA, Badry AS, Abdulsada AK. The use of Urinary α-amylase level in a diagnosis of Chronic renal failure. Res J Pharm Technol. 2021; 14(3):1597-1600. http://dx.doi.org/10.5958/0974- 360X.2021.00283.3
• [26] Abdulhussein HA, Alwasiti EA, Khiro NK, NILE AK. The potential impact of vascular endothelial growth factor rs699947 polymorphisms on breast tumors susceptibility in a sample of Iraqi females. Acta Pharm Sci. 2024;62(2):268- 277. http://dx.doi.org/10.23893/1307.APS6217.