Extracellular Vesicles of Porphyromonas gingivalis Disrupt Trophoblast Cell Interaction with Vascular and Immune Cells in an In Vitro Model of Early Placentation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Porphyromonas gingivalis Outer Membrane Vesicle Isolation
2.2. Trophoblast, THP-1 Monocyte, and Endothelial Cell Cultures
2.3. Confocal Microscopy
2.4. Trophoblast-Conditioned Media Preparation
2.5. Neutrophil Isolation
2.6. Neutrophil Migration Assay
2.7. Reactive Oxygen Species (ROS) Production
2.8. qPCR
2.9. Antioxidant Enzyme Activity
2.10. MTT Assay
2.11. Endothelial Cell Migration
2.12. Monocyte Adhesion Assay
2.13. Statistical Analysis
3. Results
3.1. Porphyromonas gingivalis Outer Membrane Vesicle Isolation and Internalization by Trophoblast Cells
3.2. Pg OMVs Modulate Trophoblast Cell Oxidative State and Unbalance Trophoblast Marker Expression
3.3. Trophoblast Cells Primed with Pg OMVs Enhance Neutrophil Chemoattraction and Activation
3.4. Trophoblast Cells Primed with Pg OMVs Impair Endothelial Cell Migration and Activation
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Sequence | |
---|---|---|
VEGF-a | Sense | CGCAGCTACTGCCATCCAAT |
Antisense | GTGAGGTTTGATCCGCATAATCT | |
PLGF | Sense | TGTTCAGCCCATCCTGTGTCTC |
Antisense | CCCAGAACGAACGGATCTTTAGG | |
IL-6 | Sense | TTCGGTACATCCTCGACGGC |
Antisense | TCACCAGGCAAGTCTCCTCA | |
IL-8 | Sense | CCAACACAGAAATTATTGTAAAGC |
Antisense | CACTGGCATCTTCACTGATTC | |
sFLT-1 | Sense | AGAGGTGAGCACTGCAACAA |
Antisense | TCTCCTCCGAGCCTGAAAGT | |
MCP-1 | Sense | CAGCAGCAAGTGTCCCAAAG |
Antisense | GAGTGAGTGTTCAAGTCTTCGG | |
TNF-α | Sense | GCCTCTTCTCCTTCCTGATCG |
Antisense | CAGCTTGAGGGTTTGCTACA | |
GP91phox | Sense | ACATTCAACCTCTGCCACCAT |
Antisense | ACCCCAGCCAAACCAGAATG | |
P47phox | Sense | CCCACGGACAACCAGACAAA |
Antisense | TCTGACAGAACCACCAACCG | |
P67phox | Sense | CTGTTTGCCTGTGAGGTGTT |
Antisense | AGACACACTCCATCGCCTTG | |
YHWAZ | Sense | CAGAGAGAAAATTGAGACGGAGC |
Antisense | GTGACTGATCGACAATCCCTTTC |
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Lara, B.; Sassot, M.; Calo, G.; Paparini, D.; Gliosca, L.; Chaufan, G.; Loureiro, I.; Vota, D.; Ramhorst, R.; Pérez Leirós, C.; et al. Extracellular Vesicles of Porphyromonas gingivalis Disrupt Trophoblast Cell Interaction with Vascular and Immune Cells in an In Vitro Model of Early Placentation. Life 2023, 13, 1971. https://doi.org/10.3390/life13101971
Lara B, Sassot M, Calo G, Paparini D, Gliosca L, Chaufan G, Loureiro I, Vota D, Ramhorst R, Pérez Leirós C, et al. Extracellular Vesicles of Porphyromonas gingivalis Disrupt Trophoblast Cell Interaction with Vascular and Immune Cells in an In Vitro Model of Early Placentation. Life. 2023; 13(10):1971. https://doi.org/10.3390/life13101971
Chicago/Turabian StyleLara, Brenda, Matías Sassot, Guillermina Calo, Daniel Paparini, Laura Gliosca, Gabriela Chaufan, Iñaki Loureiro, Daiana Vota, Rosanna Ramhorst, Claudia Pérez Leirós, and et al. 2023. "Extracellular Vesicles of Porphyromonas gingivalis Disrupt Trophoblast Cell Interaction with Vascular and Immune Cells in an In Vitro Model of Early Placentation" Life 13, no. 10: 1971. https://doi.org/10.3390/life13101971