(1184) Trophoblast injury post in vitro percussion trauma

Objective:
Trauma is the leading cause of non-obstetrical maternal death and is associated with adverse pregnancy outcomes, including placental abruption and fetal demise.   Although placental injury is a recognized consequence of maternal trauma, its direct effects on trophoblast function are not well understood. This study aims to investigate the cellular effects of percussion trauma on human trophoblasts in vitro.

Study Design:
Primary cultures of human placental extravillous trophoblasts (HTR-8/SVneo) were subjected to fluid percussion trauma (FPI, 5 atm pressure; n=6 Control; n= 6 Trauma), and were analyzed 48 h after trauma, which had been validated in our traumatic brain injury models. Cell viability was assessed with a fluorometric assay. Immunofluorescence and Western blot analyses were performed to measure the levels of mitochondrial markers (Translocase of the Outer Mitochondrial membrane (TOM20), Cytochrome C Oxidase subunit 1 (COX1), Voltage-Dependent Anion Channel 1 (VDAC1), Succinate Dehydrogenase Complex subunit A (SDHA) and the antiangiogenic factor Soluble Fms-Like Tyrosine kinase-1 (sFlt-1). Data were analyzed using the Mann-Whitney test.

Results:
Trophoblasts exposed to trauma showed a 30% decrease in cell viability compared to controls (p < 0.0043) (Fig. 1 A & B). Immunofluorescence revealed significantly higher levels of sFlt-1 (Fig. 1C) and SDHA (Fig. 1D) (42% and 59%, respectively; p = 0.001) in the injured cells, indicating a cellular stress response. Western blot analysis showed a twofold decrease in TOM20 levels (53%, p< 0.001) and a slight reduction in VDAC1 (9%), while COX1 levels remained unchanged (Fig. 2 A-D).

Conclusion:
Mechanical trauma leads to significant mitochondrial changes and cellular stress in human trophoblasts, evidenced by decreased viability, reduced expression of TOM20 and VDAC1, and increased levels of sFlt-1 and SDHA, indicating altered anti-angiogenic signaling. These patterns are similar to those observed in hypoxia-related placental disorders. This may explain how percussion trauma contributes to adverse pregnancy outcomes.