Assessment of apoptosis and appearance of hepatocyte growth factor in placenta at different gestational ages: A cross-sectional study


Background: Fetal growth is determined by the interaction between mother and fetus using the placental interface throughout the pregnancy.

Objective: To research apoptosis and appearance of hepatocyte growth factor (HGF) in placentas of different gestational ages and to describe the anthropometrical and clinical indices of mothers and newborns.

Materials and Methods: The study material was obtained from 53 human immunodeficiency virus negative pregnant women of legal age without systemic diseases. The staining of placental apoptotic cells was processed by a standard in situ cell death detection kit. The detection of HGF was provided by the ImmunoCruz goat ABC Staining System protocol sc-2023. Relative distribution of positive structures was evaluated using the semiquantitative counting method.

Results: The mean rank value of the amount of HGF-containing cells (cytotrophoblasts, syncytiotrophoblasts, extravillous trophoblasts, Höfbauer cells, and cells of extraembryonic mesoderm) was 1.61 ± 0.94. Apoptotic cells (cytotrophoblasts, syncytiotrophoblasts, extravillous trophoblasts, and cells of extraembryonic mesoderm) were found in all placental samples of various gestational ages (term 13.00 ± 13.05 and preterm 27.00 ± 18.25); in general, their amount decreased with advancing gestational age of the placenta (p < 0.01).

Conclusion: Weight of a placenta directly depends on the gestational age and correlates with the main fetal anthropometrical parameters (weight, length, and head and chest circumferences). The decrease in HGF-containing and apoptotic cells with advancing gestation depends on the adaptation potential of the placenta, proving the other ways of cellular disposition.

Key words: Pregnancy, Placenta, Gestational age, Apoptosis, Immunohistochemistry.

[1] Pritchard NL, Hiscock RJ, Lockie E, Permezel M, McGauren MFG, Kennedy AL, et al. Identification of the optimal growth charts for use in a preterm population: An Australian state-wide retrospective cohort study. PLoS Med 2019; 16: e1002923.

[2] Sharp AN, Heazell AEP, Crocker IP, Mor G. Placental apoptosis in health and disease. Am J Reprod Immunol 2010; 64: 159–169.

[3] Zhao W, Shen WW, Cao XM, Ding WY, Yan LP, Gao LJ, et al. Novel mechanism of miRNA-365-regulated trophoblast apoptosis in recurrent miscarriage. J Cell Mol Med 2017; 21: 2412–2425.

[4] Jerzak M, Bischof P. Apoptosis in the first trimester human placenta: The role in maintaining immune privilege at the maternal–foetal interface and in the trophoblast remodeling. Eur J Obstet Gynecol Reprod Biol 2002; 100: 138–142.

[5] Smith SC, Baker PN, Symonds EM. Placental apoptosis in normal human pregnancy. Am J Obstet Gynecol 1997; 177: 57–65.

[6] Huppertz B, Kingdom JC. Apoptosis in the trophoblast-role of apoptosis in placental morphogenesis. J Soc Gynecol Investig 2004; 11: 353–362.

[7] Stenhause C, Hogg CO, Achworth ChJ. Associations between fetal size, sex and both proliferation and apoptosis at the porcine feto-maternal interface. Placenta 2018; 70: 15–24.

[8] Straszewski-Chavez SL, Abrahams VM, Mor G. The role of apoptosis in the regulation of trophoblast survival and differentiation during pregnancy. Endocr Rev 2005; 26: 877–897.

[9] Knöfler M, Haider S, Saleh L, Pollheimer J, Gamage TKJB, James J. Human placenta and trophoblast development: Key molecular mechanisms and model systems. Cell Mol Life Sci 2019; 76: 3479– 3496.

[10] Bolnick AD, Bolnick JM, Kohan-Ghard HR, Kilburn BA, Hertz M, Dai J, et al. Nifedipine prevents apoptosis of alvohol-exposed first trimester trophoblast cells. Alcohol Clin Exp Res 2018; 42: 53–60.

[11] Lumicisi BA, Cartwright JE, Leslie K, Wallace AE, Whitley GS. Inhibition of DDAH1, but not DDAH2, results in apoptosis of a human trophoblast cell line in response to TRAIL. Hum Reprod 2015; 30: 1813–1819.

[12] Chaudhary P, Babu GS, Sobti RC, Gupta SK. HGF regulate HTR-8/SVneo trophoblastic cells migration/invasion under hypoxic conditions through increased HIF-1α expression via MAPK and PI3K pathways. J Cell Commun Signal 2019; 13: 503–521.

[13] Travaglino A, Raffone A, Saccone G, Migliorini S, Maruotti GM, Esposito G, et al. Placental morphology, apoptosis, angiogenesis and epithelial mechanisms in early-onset preeclampsia. Eur J Obstet Gynecol Reprod Biol 2019; 234: 200–206.

[14] Stefanini M, De Martino C, Zamboni L. Fixation of ejaculated spermatozoa for electron microscopy. Nature 1967; 216: 173–174.

[15] Negoescu A, Guillermet C, Lorimier P, Brambilla E, Labat-Moleur F. Importance of DNA fragmentation in apoptosis with regard to TUNEL specificity. Biomed Pharmacother 1998; 13: 81–86.

[16] Pilmane M, Rumba I, Sundler F, Luts A. [Patterns of distribution and occurrence of neuroendocrine elements in lungs of humans with chronic lung diseases.] Proc Latv Acad Sci Sect B 1998; 52: 144– 152. (in Latvian)

[17] Riffenburgh RH. Tests on ranked data. In: Riffenburgh RH, editor. Statistics in medicine. 3th Ed. California: Elsevier Inc.; 2012: 221–248.

[18] Riffenburgh RH. Regression and correlation. In: Riffenburgh RH. Statistics in medicine. 3th Ed. California: Elsevier Inc.; 2012: 443–472.

[19] Hutcheon JA, McNamara H, Platt RW, Benjamin A, Kramer MS. Placental weight for gestational age and adverse perinatal outcomes. Obstet Gynecol 2012; 119: 1251–1258.

[20] Salavati N, Smies M, Ganzevoort W, Charles AK, Erwich JJ, Plösch T, et al. The possible role of placental morphometry in the detection of fetal growth restriction. Front Physiol 2018; 9: 1884. 1–12.

[21] Ouyang F, Parker M, Cerda S, Pearson C, Fu L, Gillman MW, et al. Placental weight mediates the effects of prenatal factors on fetal growth: The extent differs by preterm status. Obesity 2013; 21: 609–620.

[22] Panti AA, Ekele BA, Nwobodo EI, Yakubu A. The relationship between the weight of the placenta and birth weight of the neonate in a Nigerian Hospital. Niger Med J 2012; 53: 980–984.

[23] Hayward CE, Lean S, Sibley CP, Jones RL, Wareing M, Greenwood SL, et al. Placental adaptation: What can we learn from birthweight: Placental weight ratio? Front Physiol 2016; 7: 28. 1–13.

[24] Souza MA, Brizot ML, Biancolin SE, Schultz R, Carvalho MHB, Francisco RPV, et al. Placental weight and birth weight to placental weight ratio in monochorionic and dichorionic growth-restricted and non-growth-restricted twins. Clinics 2017; 72: 265–271.

[25] Lemas DJ, Briton JT, Shapiro ALB, Glueck DH, Friedman JE, Dabelea D. Associations of maternal weight status prior and during pregnancy with neonatal cardiometabolic markers at birth: The Healthy Start study. Int J Obes 2015; 39: 1437–1442.

[26] Matsuda Y, Itoh T, Itoh H, Ogawa M, Sasaki K, Kanayama N, et al. Impact of placental weight and fetal/placental weight ratio Z score on fetal growth and the perinatal outcome. Int J Med Sci 2018; 15: 484–491.

[27] Somerset DA, Jauniaux E, Strain AJ, Afford S, Kilby MD. Hepatocyte growth factor concentration in maternal and umbilical cord blood samples and expression in fetal liver. J Soc Gynecol Investig 2000; 7: 333–337.

[28] López-Valladares MJ, Teresa Rodríguez-Ares MT, Touriño R, Gude F, Silva MT, Couceiro J. Donor age and gestational age influence on growth factor levels in human amniotic membrane. Acta Ophthalmol 2010; 88: 211–216.

[29] Ueno M, Lee LK, Chhabra A, Kim YJ, Sasidharan R, Handel B, et al. c-Met-dependent multipotent labyrinth trophoblast progenitors establish placental exchange interface. Dev Cell 2013; 27: 373–386.

[30] Dai Ch, Saleem MA, Holzman LB, Mathieson P, Liu Y. Hepatocyte growth factor signaling ameliorates podocyte injury and proteinuria. Kidney Int 2010; 77: 962–973.

[31] Mungunsukh O, McCart EA, Day RM. Hepatocyte growth factor isoforms in tissue repair, cancer, and fibrotic remodeling. Biomedicines 2014; 2: 301–326.

[32] Duszyc K, Gomez GA, Schroder K, Sweet MJ, Yapa AS. In life there is death: How epithelial tissue barriers are preserved despite the challenge of apoptosis. Tissue Barriers 2017; 5: e1345353. 1–12.

[33] Zhang S, Regnault TRH, Barker PL, Botting KJ, McMillen IC, McMillan CM, et al. Placental adaptations in growth restriction. Nutrients 2015; 7: 360–389.

[34] Athapathu H, Jayawardana MAJ, Senanayaka L. A study of the incidence of apoptosis in the human placental cells in the last weeks of pregnancy. J Obstet Gynecol 2003; 23: 515–517.

[35] Longtine MS, Chen B, Odibo AO, Zhong Y, Nelson DM. Villous trophoblast apoptosis is elevated and restricted to cytotrophoblasts in pregnancies complicated by preeclampsia, IUGR, or preeclampsia with IUGR. Placenta 2012; 33: 352–359.

[36] Longtine MS, Chen B, Odibo AO, Zhong Y, Nelson DM. Caspase-mediated apoptosis of trophoblasts in term human placental villi is restricted to cytotrophoblasts and absent from the multinucleated syncytiotrophoblast. Reproduction 2012; 143: 107–121.

[37] Sharma M, Kumar R, Bhatla N, Dhingra R. A comparative study of apoptosis in placentas of normal and preeclamptic Indian pregnant women by TUNEL assay and M30 immunostaining. J Clin Lab Anal 2012; 26: 459–466.

[38] Szpilbarg N, Castro-Parodi M, Reppetti J, Repetto M, Maskin B, Martinez N, et al. Placental programmed cell death: Insights into the role of aquaporins. Mol Hum Reprod 2016; 22: 46–56.

[39] Vogt Isaksen C. Maternal smoking, intrauterine growth restriction, and placental apoptosis. Pediatr Dev Pathol 2004; 7: 433–442.

[40] Kadyrov M, Schmitz C, Black S, Kaufmann P, Huppertz B. Pre-eclampsia and maternal anaemia display reduced apoptosis and opposite invasive phenotypes of extravillous trophoblast. Placenta 2003; 24: 540–548.

[41] Ding J, Yin T, Yan N, Cheng Y, Yang J. FasL on decidual macrophages mediates trophoblast apoptosis: A potential cause of recurrent miscarriage. Int J Mol Med 2019; 43: 2376–2386.

[42] Gong JS, Kim GJ. The role of autophagy in the placenta as a regulator of cell death. Clin Exp Reprod Med 2014; 41: 97–107.