Porphyromonas gingivalis-mediated disruption in spiral artery remodeling is associated with altered uterine NK cell populations and dysregulated IL-18 and Htra1

Abstract

Impaired spiral artery remodeling (IRSA) underpins the great obstetrical syndromes. We previously demonstrated that intrauterine infection with the periodontal pathogen, Porphyromonas gingivalis, induces IRSA in rats. Since our previous studies only examined the end stage of arterial remodeling, the aim of this study was to identify the impact of P. gingivalis infection on the earlier stages of remodeling. Gestation day (GD) 11 specimens, a transition point between trophoblast-independent remodeling and the start of extravillous trophoblast invasion, were compared to late stage GD18 tissues. P. gingivalis was found in decidual stroma of GD11 specimens that already had reduced spiral artery remodeling defined as smaller arterial lumen size, increased retention of vascular smooth muscle, and decreased invasion by extravillous trophoblasts. At GD11, P. gingivalis-induced IRSA coincided with altered uterine natural killer (uNK) cell populations, decreased placental bed expression of interleukin-18 (IL-18) with increased production of temperature requirement A1 (Htra1), a marker of oxidative stress. By GD18, placental bed IL-18 and Htra1 levels, and uNK cell numbers were equivalent in control and infected groups. However, infected GD18 placental bed specimens had decreased TNF + T cells. These results suggest disturbances in placental bed decidual stroma and uNK cells are involved in P. gingivalis-mediated IRSA.

Differential affinity chromatography reveals a link between Porphyromonas gingivalis–induced changes in vascular smooth muscle cell differentiation and the type 9 secretion system

“…Therefore, we sought to determine if AoSMCs could serve as a biologically relevant surrogate for spiral arterial VSMCs since women diagnosed with IRSA exhibit systemic arterial dysfunction during pregnancy that persists after giving birth ( Hermes et al., 2013 ; Veerbeek et al., 2015 ; Harris et al., 2019 ). Moreover, our rat model of P. gingivalis –induced IRSA is created by a chronic oral infection of 3 months duration before breeding ( Tavarna et al., 2020 ; Tavarna et al., 2022 ), which could promote systemic maternal vascular injury. The aorta from sham-inoculated controls and P. gingivalis A7UF-induced IRSA-affected dams were evaluated for vascular injury.…”

“…Another limitation of our study is that we used one set of bacterial culture conditions that would not reflect the full repertoire of P. gingivalis proteomes that change with the composition of the growth medium ( Veith et al., 2018 ). However, an important caveat of our approach was that it was designed to mimic conditions that we have observed in our in vivo model of infection ( Phillips et al., 2018 ; Tavarna et al., 2020 ; Tavarna et al., 2022 ). Despite the limitations of our proteome study, we identified several P. gingivalis proteins in IRSA-inducing A7UF that had strong, stable interactions with AoSMC proteins that did not occur with non-IRSA-inducing W83.…”

“…Bacteria for in vitro infection and for proteomics were prepared from cultures that were 48 h old. Bacterial colonies from agar were chosen to approximate bacterial conditions in tissue biofilms that we have observed in placental bed specimens from infected animals ( Tavarna et al., 2020 ; Tavarna et al., 2022 ).…”

“…A select panel of genes was evaluated using the following Quantitect primers from Qiagen Inc. (Germantown, MD, USA): b-actin, Actb as the reference gene (catalog# QT00193473), interleukin-1b, Il1b (Qiagen catalog# QT00181657), the tumor necrosis factor, Tnf (Qiagen catalog# QT00178717), chemokine monocyte chemotactic protein (Mcp1/ Ccl2) [forward:5'-ATGAGTCGGCTGGAGAACTA-3', reverse: 5'ACTTCTGGACCCATTCCTTATTG-3'], Tgfb1 (Qiagen catalog# QT00187796), endoglin, Eng [forward: 5'-GCGTCCTT GTTCCAAACATTC-3', reverse 5'-ACAGCAAGAAGAGG CAAGAG-3'], e-Selectin (Sele) [forward:5'-GTATCCAT CCATCCCACAGAAG-3', reverse: 5'-CAGTTGTGTCCAC TCGATAGTT-3'], Icam-1, Icam1 [forward: 5'-GTATCCATCCA TCCCACAGAA-3', reverse: 5'-CAGTTGTGTCCACTCGAT AGTT-3', matrix metalloproteinase 2, Mmp2 [forward: 5'-TGGACTGGAGAAGGACAA-3', reverse: 5'-CTGCTGTATT CCCGACCATTA-3'], matrix metalloproteinase 9, Mmp9 [forward: 5'-CCCAACCTTTACCAGCTACTC-3', reverse: 5'-GTCAGAACCGACCCTACAAAG-3"], and the tissue inhibitor of metalloproteinase-1, Timp1[forward:5'-TGGCATCC TCTTGTTGCTATC-3', reverse: 5'-CCTTATAACCAGGT CCGAGTTG-3']. Real-time qPCR was performed with the Light Cycler ® 96 real-time PCR System (Roche Applied Science, Indianapolis, IN, USA) as previously described (Tavarna et al, 2022).…”

“…Porphyromonas gingivalis is implicated in a variety of obstetric complications including early pregnancy loss, early-onset preeclampsia with and without fetal growth restriction, preterm labor, and the preterm premature rupture of membranes ( Barak et al., 2007 ; León et al., 2007 ; Swati et al., 2012 ; Chaparro et al., 2013 ; Ibrahim et al., 2015 ; Vanterpool et al., 2016 ). Using a rat model of chronic periodontitis, we previously demonstrated that intrauterine colonization by various strains of P. gingivalis impairs the physiologic remodeling of the uterine spiral arteries (IRSA) during pregnancy ( Phillips et al., 2018 ; Tavarna et al., 2020 ; Tavarna et al., 2022 ). Since IRSA underlies the major obstetrical syndromes, this evidence provided a common mechanistic link whereby P. gingivalis could promote such a diverse array of pregnancy complications.…”