195. Phenotype restoration of trophoblasts in preeclampsia by Low Level Light Therapy (LLLT)

In our previous work we demonstrated the importance of altered extracellular matrix (ECM) in disease processes and how treatment with 670 nm LLLT can restore cell behavior. Of this study focuses on the signaling properties of altered ECM in preeclampsia, its contribution to preeclampsia, and the possibility of a novel therapeutic modality restoring the trophoblast phenotype with LLLT. To study signaling pathways we decellularized placentas from healthy and preeclampsic subjects with 1%SDS followed by triton X and thorough wash steps. First trimester human trophoblasts (HTG8) were cultured on these two different matrices. This allows us to study trophoblast behavior in their proper microenvironment and follow the trophoblast phenotype changes. Immunohistochemistry and western blot analysis were performed with a focus on PLGF, ERRg, FoxP3, and HLA-G expression comparing the different matrices. Readouts are fluorescence intensity and western blot density in arbitrary units (AU). Furthermore, 670 nm light treatment was applied to HTG8s at an energy level of 4 J/ cm[b] once and cellular changes were studied 24hours later. Our results show that the expression of PLGF in HTG8s was decreased from 102AU to 68 AU comparing culture on control matrix to preeclampsic matrix. ERRg expression was significantly reduced in HTG8s from 80.33AU to 18.6AU. FoxP3 expression was decreased from109.38AU to 82.1AU. HLA-G expression was significantly decreased from 105.94 to 49.48AU. LLLT restored the phenotype of HTG8s cultured on preeclampsic matrix to the phenotype of HTG8s cultured on control placenta. Statistics were performed using the Student’s t-test. Our data demonstrate for the first time that the ECM of the placenta and therefore the microenvironment has great impact on trophoblast signaling. Preeclampsic ECM alters the trophoblast phenotype and LLLT restored the phenotype to the phenotype found in trophoblasts cultured on control ECM.