Impact of Prematurity on the Buccal Epithelial Cells of the Neonates via Wnt/Beta-Catenin Signaling Pathway and Apoptosis

Abstract

Objective Understanding the reflections of prematurity is necessary for the management of neonatal complications. We focused on the impact of prematurity and related “maternal risk factors/obstetric complications” on buccal cells of the neonates via evaluation of the Wnt/β-catenin signaling pathway and apoptosis.

Study Design This study consisted of “early preterm neonates (EPN) (≤34th gestational week [gw]) (n = 36),” “late preterm neonates (LPN) (34th– < 37th gw) (n = 46),” and “term neonates (control) (≥37th gw) (n = 56).” Cohort was also subclassified according to the presence of maternal risk factors, obstetric complications, and neonatal complications. Wnt/β-catenin signaling and caspase-3 activation pathways were studied immunocytochemically.

Results Wnt/β-catenin signaling positivity was statistically more frequent at buccal smears of the EPN and LPN groups compared with controls (p < 0.001). The cutoff for gestational age at delivery in receiver operating characteristic curve with the best balance of sensitivity (67.4%) and specificity (67.3%) was 35.8th gw for determining the reduction of Wnt/β-catenin signaling positivity (p < 0.001). The study demonstrated that obstetric complications significantly affected the activity of signaling, while maternal risk factors do not have any effect on Wnt/β-catenin signaling pathway (p = 0.003 and p = 0.828, respectively). This study also demonstrated a significant relationship between Wnt/β-catenin signaling pathway and the presence of neonatal complications (p = 0.015).

Conclusion Dynamic characteristics of buccal cells are influenced by prematurity and related obstetric and neonatal problems. Buccal smear is a good tool to investigate the impact of prematurity and obstetric problems on perinatal outcome.

Key Points

• Neonatal buccal cells are affected by prematurity and related obstetric/neonatal problems.

• 35.8th gw is critical for determining the reduction of Wnt/β-catenin signaling positivity.

• Obstetric and neonatal complications significantly related to Wnt/β-catenin signaling activity.

Keywords

pregnancy - preterm birth - buccal smear - Wnt/β-catenin signaling pathway - apoptosis - caspase-3 - perinatal outcome

Authors' Contribution

H.G.D. contributed in study design, experiments, data analysis, statistical analysis, and writing; H.T.C. and S.Y. in study design and sample collection; G.K. in sample collection; A.T. in study design, statistical analysis, and literature research; M.C. in statistical analysis and literature research; and M.Y., A.N.C., and M.S.B. in study design and critical reading.

Ethical Approval

The study was approved by the Hacettepe University Ethics Committee with reference number GO18/495-43. The study was based in accordance with the Declaration of Helsinki.

Data Availability Statement

Data sets are available from the corresponding author on reasonable request.

Publikationsverlauf

Eingereicht: 10. September 2021

Angenommen: 01. November 2021

Artikel online veröffentlicht:
10. Dezember 2021

• References

• 1 Turgal M, Gumruk F, Karaagaoglu E, Beksac MS. Methylenetetrahydrofolate Reductase Polymorphisms and Pregnancy Outcome. Geburtshilfe Frauenheilkd 2018; 78 (09) 871-878
• 2 Tanacan A, Fadiloglu E, Beksac MS. The importance of proteinuria in preeclampsia and its predictive role in maternal and neonatal outcomes. Hypertens Pregnancy 2019; 38 (02) 111-118
• 3 Parikh LI, Reddy UM, Männistö T. et al. Neonatal outcomes in early term birth. Am J Obstet Gynecol 2014; 211 (03) 265.e1-265.e11
• 4 Theda C, Hwang SH, Czajko A, Loke YJ, Leong P, Craig JM. Quantitation of the cellular content of saliva and buccal swab samples. Sci Rep 2018; 8 (01) 6944
• 5 Thomas P, Holland N, Bolognesi C. et al. Buccal micronucleus cytome assay. Nat Protoc 2009; 4 (06) 825-837
• 6 Odintsova VV, Hagenbeek FA, Suderman M. et al. DNA methylation signatures of breastfeeding in buccal cells collected in mid-childhood. Nutrients 2019; 11 (11) 2804
• 7 de Vries TW, Ajubi N, Slomp J, Storm H. Analyzing DNA from buccal cells is a reliable method for the exclusion of cystic fibrosis. Results of a pilot study. Genet Med 2006; 8 (03) 175-177
• 8 van Dongen J, Ehli EA, Jansen R. et al. Genome-wide analysis of DNA methylation in buccal cells: a study of monozygotic twins and mQTLs. Epigenetics Chromatin 2018; 11 (01) 54
• 9 Donmez HG, Celik HT, Kayki G. et al. Impact of preterm birth on the cellular characteristics of neonatal buccal cells. Cytopathology 2021; 32 (05) 660-670
• 10 Donmez HG, Demirezen Ş, Beksaç MS. Detection of the Wnt/beta-catenin signaling activity by using immunocytochemical technique in cervical smears. Gynecol Obstet Reprod Med 2013; 19: 91-95
• 11 Zhao C, Yu Y, Zhang Y. et al. β-Catenin controls the electrophysiologic properties of skeletal muscle cells by regulating the α2 isoform of Na+/K+-ATPase. Front Neurosci 2019; 13: 831
• 12 Shang S, Hua F, Hu ZW. The regulation of β-catenin activity and function in cancer: therapeutic opportunities. Oncotarget 2017; 8 (20) 33972-33989
• 13 Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35 (04) 495-516
• 14 Donmez HG, Demirezen S, Beksac MS. The relationship between beta-catenin and apoptosis: a cytological and immunocytochemical examination. Tissue Cell 2016; 48 (03) 160-167
• 15 Beksac MS, Tanacan A, Ozten G, Cakar AN. Low-dose low-molecular-weight heparin prophylaxis against obstetrical complications in pregnancies with metabolic and immunological disorder-associated placental inflammation. J Matern Neonatal Med 2020; 1-8
• 16 Cox RT, Kirkpatrick C, Peifer M. Armadillo is required for adherens junction assembly, cell polarity, and morphogenesis during Drosophila embryogenesis. J Cell Biol 1996; 134 (01) 133-148
• 17 Steinhart Z, Angers S. Wnt signaling in development and tissue homeostasis. Development 2018; 145 (11) dev146589
• 18 Miller LAD, Smith AN, Taketo MM, Lang RA. Optic cup and facial patterning defects in ocular ectoderm β-catenin gain-of-function mice. BMC Dev Biol 2006; 6: 14
• 19 De Langhe SP, Reynolds SD. Wnt signaling in lung organogenesis. Organogenesis 2008; 4 (02) 100-108
• 20 Abrahams VM, Chamley LW, Salmon JE. Emerging treatment models in rheumatology: antiphospholipid syndrome and pregnancy: pathogenesis to translation. Arthritis Rheumatol 2017; 69 (09) 1710-1721
• 21 Swadzba J, Iwaniec T, Musial J. Increased level of tumor necrosis factor-α in patients with antiphospholipid syndrome: marker not only of inflammation but also of the prothrombotic state. Rheumatol Int 2011; 31 (03) 307-313
• 22 Roubert A, Gregory K, Li Y. et al. The influence of tumor necrosis factor-α on the tumorigenic Wnt-signaling pathway in human mammary tissue from obese women. Oncotarget 2017; 8 (22) 36127-36136
• 23 Sulistyowati S, Zakia Y, Khasan S. High MMP-9 and TNF-α expression increase in preterm premature rupture of membranes. Universa Med 2016; 35: 33
• 24 Woods Jr. JR. Reactive oxygen species and preterm premature rupture of membranes-a review. Placenta 2001; 22 suppl A: S38-S44
• 25 Ma B, Hottiger MO. Crosstalk between Wnt/β-catenin and NF-κB signaling pathway during inflammation. Front Immunol 2016; 7: 378
• 26 Korswagen HC. Regulation of the Wnt/β-catenin pathway by redox signaling. Dev Cell 2006; 10 (06) 687-688
• 27 Unal C, Karatas E, Fadıloglu E, Portakal O, Beksac MS. Comparison of term and preterm labor procalcitonin and leukocyte cell volume, conductivity and light scatter (VCS) parameters in order to demonstrate the impact of inflammation on the triggering mechanisms of preterm uterin contractions. J Obstet Gynaecol Res 2020; 46 (05) 694-698
• 28 Huang P, Zhou Q, Lin Q. et al. Complement C3a induces axonal hypomyelination in the periventricular white matter through activation of WNT/β-catenin signal pathway in septic neonatal rats experimentally induced by lipopolysaccharide. Brain Pathol 2020; 30 (03) 495-514
• 29 Smith AK, Kilaru V, Klengel T. et al. DNA extracted from saliva for methylation studies of psychiatric traits: evidence tissue specificity and relatedness to brain. Am J Med Genet B Neuropsychiatr Genet 2015; 168B (01) 36-44
• 30 Braun PR, Han S, Hing B. et al. Genome-wide DNA methylation comparison between live human brain and peripheral tissues within individuals. Transl Psychiatry 2019; 9 (01) 47
• 31 Donmez HG. β-Catenin immunocytochemical reactivity in cervicovaginal smears during regular menstrual cycles. Asian Biomed 2020; 14: 187-194
• 32 Huppertz B, Frank HG, Reister F, Kingdom J, Korr H, Kaufmann P. Apoptosis cascade progresses during turnover of human trophoblast: analysis of villous cytotrophoblast and syncytial fragments in vitro. Lab Invest 1999; 79 (12) 1687-1702