Human amniotic epithelial cell transplantation improves scar remodeling in a rabbit model of acute vocal fold injury: a pilot study

Abstract

Objective To gain insight into the molecular mechanisms underlying the early stages of vocal fold extracellular matrix (ECM) remodeling after a mid-membranous injury resulting from the use of human amniotic epithelial cells (hAEC), as a novel regenerative medicine cell-based therapy. Methods Vocal folds of six female, New Zealand White rabbits were bilaterally injured. Three rabbits had immediate bilateral direct injection of 1 × 106 hAEC in 100 µl of saline solution (hAEC) and three with 100 µl of saline solution (controls, CTR). Rabbits were euthanized 6 weeks after injury. Proteomic analyses (in-gel trypsin protein digestion, LC–MS/MS, protein identification using Proteome Discoverer and the Uniprot Oryctolagus cuniculus (Rabbit) proteome) and histological analyses were performed. Results hAEC treatment significantly increased the expression of ECM proteins, elastin microfibril interface-located protein 1 (EMILIN-1) and myocilin that are primarily involved in elastogenesis of blood vessels and granulation tissue. A reactome pathway analysis showed increased activity of the anchoring fibril formation by collagen I and laminin, providing mechanical stability and activation of cell signaling pathways regulating cell function. hAEC increased the abundance of keratin 1 indicating accelerated induction of the differentiation programming of the basal epithelial cells and, thereby, improved barrier function. Lastly, upregulation of Rab GDP dissociation inhibitor indicates that hAEC activate the vesicle endocytic and exocytic pathways, supporting the exosome-mediated activation of cell–matrix and cell-to-cell interactions. Conclusions This pilot study suggests that injection of hAEC into an injured rabbit vocal fold favorably alters ECM composition creating a microenvironment that accelerates differentiation of regenerated epithelium and promotes stabilization of new blood vessels indicative of accelerated and improved repair.

Rat liver extracellular matrix and perfusion bioreactor culture promote human amnion epithelial cell differentiation towards hepatocyte-like cells

“…The human AEC isolation was performed as previously described. [30][31][32] Briefly, the amnion membrane was removed from the inner layer of the placenta from full-term healthy pregnancies, and washed extensively to remove blood. The amnion membrane was immersed in TrypLE 10x (Gibco) for 30 min at 37°C to release mainly AEC from the amnion membrane.…”

Human Amniotic Epithelial Cells Promote Chx10−/Pax6+Müller Glia Subpopulation Reprogramming into Photoreceptor-like Cells

“…In addition, hAECs have the unique capacity to regulate the extracellular microenvironment, especially the extracellular matrix (ECM), when compared with other stem cells. hAECs improve tissue repair and regeneration by regulating ECM remodeling during lung fibrosis and premature ovarian failure 26,27 . Previously, photoreceptors differentiated from hAECs were transplanted into the retina to improve visual function in RD rats 28 .…”

eEF2 improves dense connective tissue repair and healing outcome by regulating cellular death, autophagy, apoptosis, proliferation and migration

“…Prior studies in animals have utilized proteomics to delineate the healing of DCT [ 5 – 8 ]. However, there is a very limited number of reports examining how the human proteome changes in this process.…”