title: Inhibition of Proliferation and Epithelial Mesenchymal Transition in Retinal Pigment Epithelial Cells by Heavy Chain-Hyaluronan/Pentraxin 3.
abstract: Proliferative vitreoretinopathy (PVR) is mediated by proliferation and epithelial mesenchymal transition (EMT) of retinal pigment epithelium (RPE). Because heavy chain-hyaluronic acid/pentraxin 3 (HC-HA/PTX3) purified from human amniotic membrane exerts anti-inflammatory and anti-scarring actions, we hypothesized that HC-HA/PTX3 could inhibit these PVR-related processes in vitro. In this study, we first optimized an ARPE-19 cell culture model to mimic PVR by defining cell density, growth factors, and cultivation time. Using this low cell density culture model and HA as a control, we tested effects of HC-HA/PTX3 on the cell viability (cytotoxicity), proliferation (EGF + FGF-2) and EMT (TGF-β1). Furthermore, we determined effects of HC-HA/PTX3 on cell migration (EGF + FGF-2 + TGF-β1) and collagen gel contraction (TGF-β1). We found both HA and HC-HA/PTX3 were not toxic to unstimulated RPE cells. Only HC-HA/PTX3 dose-dependently inhibited proliferation and EMT of stimulated RPE cells by down-regulating Wnt (β-catenin, LEF1) and TGF-β (Smad2/3, collagen type I, α-SMA) signaling, respectively. Additionally, HA and HC-HA/PTX3 inhibited migration but only HC-HA/PTX3 inhibited collagen gel contraction. These results suggest HC-HA/PTX3 is a non-toxic, potent inhibitor of proliferation and EMT of RPE in vitro, and HC-HA/PTX3's ability to inhibit PVR formation warrants evaluation in an animal model.
authors.affiliation: TissueTech, Inc., Miami, FL, 33173, USA.
authors.forename: Hua
authors.initials: H
authors.lastname: He
authors.affiliation: Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA.|Flaum Eye Institute, University of Rochester Medical Center, Rochester, NY, 14642, USA.
authors.forename: Ajay E
authors.initials: AE
authors.lastname: Kuriyan
authors.affiliation: TissueTech, Inc., Miami, FL, 33173, USA.
authors.forename: Chen-Wei
authors.initials: CW
authors.lastname: Su
authors.affiliation: TissueTech, Inc., Miami, FL, 33173, USA.
authors.forename: Megha
authors.initials: M
authors.lastname: Mahabole
authors.affiliation: Ocular Surface Center and Ocular Surface Research &Education Foundation, Miami, FL, 33173, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: TissueTech, Inc., Miami, FL, 33173, USA.
authors.forename: Ying-Ting
authors.initials: YT
authors.lastname: Zhu
authors.affiliation: Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA.
authors.forename: Harry W
authors.initials: HW
authors.lastname: Flynn
authors.affiliation: Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA.
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Parel
authors.affiliation: TissueTech, Inc., Miami, FL, 33173, USA.|Ocular Surface Center and Ocular Surface Research &Education Foundation, Miami, FL, 33173, USA.
authors.forename: Scheffer C G
authors.initials: SC
authors.lastname: Tseng
mesh_terms: D015415:Biomarkers; D002097:C-Reactive Protein / Q000494:pharmacology*; D002465:Cell Movement; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D021381:Protein Transport; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism*; D000683:Serum Amyloid P-Component / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism / Q000494:pharmacology; D018630:Vitreoretinopathy, Proliferative / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D000099153:Pentraxins
chemical_list: D015415:Biomarkers; D002097:C-Reactive Protein; D004815:Epidermal Growth Factor; D006820:Hyaluronic Acid; D000683:Serum Amyloid P-Component; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; D000099153:Pentraxins