title: Nerve regeneration by human corneal stromal keratocytes and stromal fibroblasts.
abstract: Laser refractive surgeries reshape corneal stroma to correct refractive errors, but unavoidably affect corneal nerves. Slow nerve regeneration and atypical neurite morphology cause desensitization and neuro-epitheliopathy. Following injury, surviving corneal stromal keratocytes (CSKs) are activated to stromal fibroblasts (SFs). How these two different cell types influence nerve regeneration is elusive. Our study evaluated the neuro-regulatory effects of human SFs versus CSKs derived from the same corneal stroma using an in vitro chick dorsal root ganglion model. The neurite growth was assessed by a validated concentric circle intersection count method. Serum-free conditioned media (CM) from SFs promoted neurite growth dose-dependently, compared to that from CSKs. We detected neurotrophic and pro-inflammatory factors (interleukin-8, interleukin-15, monocyte chemoattractant protein-1, eotaxin, RANTES) in SFCM by Bio-Plex Human Cytokine assay. More than 130 proteins in SFCM and 49 in CSKCM were identified by nanoLC-MS/MS. Proteins uniquely present in SFCM had reported neuro-regulatory activities and were predicted to regulate neurogenesis, focal adhesion and wound healing. Conclusively, this was the first study showing a physiological relationship between nerve growth and the metabolically active SFs versus quiescent CSKs from the same cornea source. The dose-dependent effect on neurite growth indicated that nerve regeneration could be influenced by SF density.
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Ophthalmology and Visual Science Academic Clinical Research Program, Duke-National University of Singapore Graduate Medical School, Singapore.
authors.forename: Gary Hin-Fai
authors.initials: GH
authors.lastname: Yam
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Cornea and External Eye Disease Service Team, Singapore National Eye Centre, Singapore.
authors.forename: Geraint P
authors.initials: GP
authors.lastname: Williams
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Melina
authors.initials: M
authors.lastname: Setiawan
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Nur Zahirah Binte M
authors.initials: NZ
authors.lastname: Yusoff
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Xiao-Wen
authors.initials: XW
authors.lastname: Lee
authors.affiliation: Ophthalmology and Visual Science Academic Clinical Research Program, Duke-National University of Singapore Graduate Medical School, Singapore.|Singapore Eye Research Institute, Singapore.
authors.forename: Hla Myint
authors.initials: HM
authors.lastname: Htoon
authors.affiliation: Ophthalmology and Visual Science Academic Clinical Research Program, Duke-National University of Singapore Graduate Medical School, Singapore.|Singapore Eye Research Institute, Singapore.|Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Department of Ophthalmology, RWTH Aachen University, Aachen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Fuest
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Ophthalmology and Visual Science Academic Clinical Research Program, Duke-National University of Singapore Graduate Medical School, Singapore.|Cornea and External Eye Disease Service Team, Singapore National Eye Centre, Singapore.|Singapore Eye Research Institute, Singapore.|School of Materials Science and Engineering, Nanyang Technological University, Singapore.
authors.forename: Jodhbir S
authors.initials: JS
authors.lastname: Mehta
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D002642:Chick Embryo; D003315:Cornea / Q000166:cytology / Q000254:growth & development*; D060527:Corneal Keratocytes / Q000166:cytology*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000166:cytology*; D006801:Humans; D020731:Keratomileusis, Laser In Situ / Q000009:adverse effects; D009416:Nerve Regeneration / Q000502:physiology*; D016501:Neurites / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology*
chemical_list: D017077:Culture Media, Conditioned