Placental basement membrane proteins are required for effective cytotrophoblast invasion in a three‐dimensional bioprinted placenta model

Abstract

Fetal cytotrophoblast invasion of maternal decidual vasculature is necessary to normal pregnancy. In preeclampsia, there is shallow invasion and abnormal remodeling of the uterine vasculature that lead to significant maternal and perinatal morbidity and mortality. The placental basement membrane (BM) proteins (e.g., laminin and collagen) has been implicated in the development of placenta while the level of laminin is significantly lower in preeclampsia. However, there are very limited studies, if any, on the effect of extracellular matrix (ECM) microenvironment on the invasion of cytotrophoblast. In this study, we hypothesized that placental BM proteins are required for effective cytotrophoblast invasion. Using proteomics, we found that more than 80% of ECM proteins in placental basal plate (pECM) were BM proteins. In addition to upregulating expressions of MMP2 (1.5-fold) and MMP9 (6.3-fold), pECM significantly increased the motility rates of cytotrophoblasts by 13-fold (from 5.60 ± 0.95 to 75.5 ± 21.8 µm/day) to achieve an effective invasion rate that was comparable to in vivo results. Treatments with PI3K inhibitors completely removed the pECM-enhanced invasive phenotypes and genotypes of cytotrophoblasts, suggesting its dominant role in cytotrophoblast-ECM interactions. Our results described, for the first time, the substantial effects of the ECM microenvironment on regulating cytotrophoblast invasion, an area that is less investigated but appear to be critical in the pathogenesis of preeclampsia. Moreover, the approach presented in this work that fabricates organ models with organ-specific ECM can be an attractive option to screen and develop novel therapeutics and biomarkers not only in preeclampsia but also other diseases such as cancer metastasis. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1476-1487, 2018.

Model Placental Barrier Phenotypic Response to Fluoxetine and Sertraline: A Comparative Study

“…Comparing EC type, the largest difference was observed in small molecule permeability, though even this difference was relatively modest; permeability was still within the same order of magnitude. In evaluating placental ECM, the tissue was decellularized to an extent similar to other studies, with low DNA content and similar protein composition . Modest differences in barrier formation based on placental ECM content were observed, which is comparable to previous findings that ECM proteins have minimal impact on permeability characteristics, albeit studied for ECs .…”

“…By comparison, the placenta‐on‐a‐chip and transwell models often lack the cellular complexity and 3D nature of the BPB . Extracellular matrix (ECM) proteins, important in placental biology, are rarely used. Similarly, how EC phenotype influences molecular permeability, with differences between macrovascular and microvascular ECs reported, rarely influences BPB model design.…”

Tuning trophoblast invasion in a gelatin hydrogel via soluble cues from the maternal-fetal interface

“…Such models can replicate relevant biophysical properties inspired by the native tissue, including matrix stiffness, extracellular matrix composition, and three-dimensional architecture. Advanced tissue engineered platforms have increasingly been utilized to study trophoblast invasion and migration in three-dimensional biomaterial platforms (8)(9)(10)(11); however, some limitations still remain with existing models. The use of three-dimensional bioprinting (8,9) and microfluidic technology (10) allows for tracking migration of trophoblast cells toward soluble factor gradients within biomaterials but these models quantify migration in dissociated cells and hence, do not recapitulate the native spherical structure of an invading blastocyst.…”

“…Advanced tissue engineered platforms have increasingly been utilized to study trophoblast invasion and migration in three-dimensional biomaterial platforms (8)(9)(10)(11); however, some limitations still remain with existing models. The use of three-dimensional bioprinting (8,9) and microfluidic technology (10) allows for tracking migration of trophoblast cells toward soluble factor gradients within biomaterials but these models quantify migration in dissociated cells and hence, do not recapitulate the native spherical structure of an invading blastocyst. Existing models that utilize embryos (11) utilize Matrigel which contains a heterogeneous combination of extracellular matrix proteins and exhibits significant lot to lot variability (12,13).…”

Development of surface functionalization strategies for 3D‐printed polystyrene constructs

“…Protein was isolated through electrophoresis, as described. Gel lanes were separated, cut into 10 equal pieces, digested with sequencing grade trypsin, and peptides extracted with acetonitrile‐formic acid buffer . Liquid‐chromatography (LC)‐MS/MS was performed using a nano‐LC system (Easy nLC1000) connected to Q Exactive mass spectrometer (ThermoFisher).…”