Potential mechanisms for chorioamniotic membrane rupture after subchorionic hematoma

Subchorionic hematoma is a risk factor for preterm prelabor rupture of membranes and preterm birth. A small proportion of persistent subchorionic hematoma leads to a chronic abruption-oligohydramnios sequence.

To determine the mechanism by which subchorionic hematomas may damage chorioamniotic membranes.

1) The number and subtype of macrophages were determined by immunohistochemistry in chorioamniotic membranes from 8 subchorionic hematoma patients who delivered preterm (25.5 (24-32) weeks of gestation (median and range)) and 6 gestational age-matched control patients (25.5 (25-28) weeks of gestation (median and range)). Further, the thickness and fibrosis of the membranes were quantified. 2) We also developed an intrauterine hematoma model in pregnant mice, and the effects of hematoma on the amnion were analyzed by histology and immunofluorescence. 3) In vitro, primary human amnion mesenchymal cells were cocultured with M2-differentiated macrophages, and changes in mesenchymal cells were analyzed.

1) Subchorionic hematoma increased the number of iron-laden macrophages in the human amnion. These macrophages were CD206⁺, a marker of macrophages required for the maintenance of homeostasis, tissue remodeling, and metabolic adaptations. The collagen layer of the amnion tended to be thickened in patients with subchorionic hematoma. Interestingly, α-smooth muscle actin⁺ myofibroblasts were increased in the amnion mesenchymal layer in patients with subchorionic hematoma. Vimentin, a mesenchymal marker, was expressed in the epithelial layer of the hematoma amnion. Together, these findings indicate epithelial-mesenchymal transition in the amnion of membranes from pregnancies with subchorionic hematomas. 2) These findings in human amnion were confirmed in a mouse model of intrauterine hematoma. 3) Further, in vitro, coculture of human amnion mesenchymal cells with M2-differentiated human macrophages resulted in transformation of these cells into α-smooth muscle actin-expressing myofibroblasts via the TGF-β‒Smad3 pathway.

Subchorionic hematoma induces migration of macrophages to chorioamniotic membranes which activate the transition of amnion mesenchymal cells to myofibroblasts. These myofibroblasts may contribute to fibrosis of the amnion and damage chorioamniotic membranes.