Tissue Glue-Based Sealing Patch for the in vivo Prevention of Iatrogenic Prelabor Preterm Rupture of Fetal Membranes.

INTRODUCTION: One of the main concerns for all fetal surgeries is the risk of preterm delivery due to the preterm prelabor rupture of the fetal membranes (iPPROM). Clinical approaches to seal fetal membrane (FM) defects are missing due to the lack of appropriate strategies to apply sealing biomaterials at the defect site. METHODS: Here, we test the performance of a previously developed strategy to seal FM defects with cyanoacrylate-based sealing patches in an ovine model up to 24 days after application. RESULTS: Patches sealed tightly the fetoscopy-induced FM defects and remained firmly attached to the defect over 10 days. At 10 days after treatment, 100% (13/13) of the patches were attached to the FMs, and 24 days after treatment 25% (1/4) of the patches placed in CO2 insufflation, and 33% (1/3) in NaCl infusion remained. However, all successfully applied patches (20/24) led to a watertight sealing at 10 or 24 days after treatment. Histological analysis indicated that cyanoacrylates induced a moderate immune response and disrupted the FM epithelium. CONCLUSION: Together, these data show the feasibility of minimally invasive sealing of FM defects by locally gathering tissue adhesive. Further development to combine this technology with refined tissue glues or healing-inducing materials holds great promise for future clinical translation.

Miniaturized Bioengineered Models for Preterm Fetal Membrane Healing.

INTRODUCTION: The reason for the absence of fetal membrane (FM) healing after a fetoscopic intervention is still unknown. We hypothesize that the lack of robust miniaturized models to study preterm FM functions is currently hampering the development of new treatments for FM healing. Specifically, miniaturized models to study preterm FM healing with minimal amounts of tissue are currently lacking. METHODS: In this study, we collected FMs from planned cesarean deliveries and developed different ex vivo models with an engineered biomaterial to study FM healing. Then, the effect of platelet-derived growth factor BB (PDGF-BB) on the migration of cells from preterm and term FMs was evaluated. RESULTS: FMs could be viably cultured ex vivo for 14 days. In a model of punctured FMs, migration of cells into FM defects was less pronounced than migration out of the tissue into the biomaterial. In a miniaturized model of preterm cell migration, PDGF-BB promoted migration of preterm amnion cells into the biomaterial. DISCUSSION AND CONCLUSION: By using a novel miniaturized model of preterm tissue, we here successfully demonstrate that PDGF-BB can promote preterm FM cell migration of microtissues encapsulated in a three-dimensional environment.

Biomaterial-based treatments for the prevention of preterm birth after iatrogenic rupture of the fetal membranes.

Minimally invasive interventions to ameliorate or correct fetal abnormalities are becoming a clinical reality. However, the iatrogenic preterm prelabor rupture of the fetal membranes (FMs) (iPPROM), which may result in preterm birth, remains a main complication. Despite the cause of iPPROM not being fully known, the puncture created by the fetoscope remains unhealed until the end of the pregnancy, which permits chorioamniotic separation and amniotic fluid leakage. Hence, there is an urgent need to develop strategies to treat the FMs after minimally invasive interventions. However, none of the previously tested strategies has been clinically translated. Here, we review the current knowledge about the FMs starting from their development and present the different models that have been developed both in vitro and ex vivo. We also systematically review and summarize the different approaches that have been investigated to plug, seal, heal or suture the FMs both in preclinical and clinical studies and discuss their limitations, outcomes, and future directions.

In vivo Sealing of Fetoscopy-Induced Fetal Membrane Defects by Mussel Glue.

INTRODUCTION: The benefits of fetal surgery are impaired by the high incidence of iatrogenic preterm prelabor rupture of the fetal membranes (iPPROM), for which chorioamniotic separation has been suggested as a potential initiator. Despite the urgent need to prevent iPPROM by sealing the fetoscopic puncture site after intervention, no approach has been clinically translated. METHODS: A mussel-inspired biomimetic glue was tested in an ovine fetal membrane (FM) defect model. The gelation time of mussel glue (MG) was first optimized to make it technically compatible with fetal surgery. Then, the biomaterial was loaded in polytetrafluoroethylene-coated nitinol umbrella-shaped receptors and applied on ovine FM defects (N = 10) created with a 10 French trocar. Its sealing performance and tissue response were analyzed 10 days after implantation by amniotic fluid (AF) leakage and histological methods. RESULTS: All ewes and fetuses recovered well after the surgery, and 100% ewe survival and 91% fetal survival were observed at explantation. All implants were tight at explantation, and no AF leakage was observed in any of them. Histological analysis revealed a mild tissue response to the implanted glue. CONCLUSION: MG showed promising properties for the sealing of FM defects and thereby the prevention of preterm birth. Studies to analyze the long-term tissue response to the sealant should be performed.

Comprehensive quantitative characterization of the human term amnion proteome.

The loss of fetal membrane (FM) integrity and function at an early time point during pregnancy can have devastating consequences for the fetus and the newborn. However, biomaterials for preventive sealing and healing of FMs are currently non-existing, which can be partly attributed to the current fragmentary knowledge of FM biology. Despite recent advances in proteomics analysis, a robust and comprehensive description of the amnion proteome is currently lacking. Here, by an optimized protein sample preparation and offline fractionation before liquid chromatography coupled to mass spectrometry (LC-MS) analysis, we present a characterization of the healthy human term amnion proteome, which covers more than 40% of the previously reported transcripts in similar RNA sequencing datasets and, with more than 5000 identifications, greatly outnumbers previous reports. Together, beyond providing a basis for the study of compromised and preterm ruptured FMs, this comprehensive human amnion proteome is a stepping-stone for the development of novel healing-inducing biomaterials. The proteomic dataset has been deposited in the ProteomeXchange Consortium with the identifier PXD019410.

Label-Free Quantification Proteomics for the Identification of Mesenchymal Stromal Cell Matrisome Inside 3D Poly(Ethylene Glycol) Hydrogels.

Cells modulate the functional properties of their environment by depositing extracellular matrix (ECM) proteins during biological processes in vivo and in vitro. Despite the ECMs central role in tissue formation, its quantification in hydrogels like Matrigel, which have a complex materials-inherent biopolymer composition is exceptionally challenging. Here, the use of protein-free, synthetic poly(ethylene glycol) hydrogels enables the analysis of deposited human bone marrow mesenchymal stromal cells ECM directly harvested from fresh 3D cell cultures by a tandem mass spectrometry (LC-MS/MS) method. In this study, it is proved that a label-free LC-MS/MS quantification method can selectively identify proteins deposited in 3D synthetic hydrogels following different growth factor (GF) treatments. Furthermore, it is shown that the sequence in which GFs are administered and the choice of stimuli significantly influences the number and abundance of ECM proteins. Therefore, this provides a versatile method to optimize GF treatments in synthetic hydrogel-based regenerative medicine and tissue engineering approaches.