title: Corneal epithelial wound healing and bactericidal effect of conditioned medium from human uterine cervical stem cells.
abstract: OBJECTIVE
  To evaluate the effect of conditioned medium from human uterine cervical stem cells (CM-hUCESCs) on corneal epithelial healing in a rat model of dry eye after alkaline corneal epithelial ulcer. We also tested the bactericidal effect of CM-hUCESCs.
  
  
  METHODS
  Dry eye was induced in rats by extraocular lacrimal gland excision, and corneal ulcers were produced using NaOH. Corneal histologic evaluation was made with hematoxylin-eosin (H&E) staining. Real-time PCR was used to evaluate mRNA expression levels of proinflammatory cytokines. We also studied the bactericidal effect of CM-hUCESCs in vitro and on infected corneal contact lenses (CLs) using Escherichia coli and Staphylococcus epidermidis bacteria. In addition, in order to investigate proteins from CM-hUCESCs that could mediate these effects, we carried out a human cytokine antibody array.
  
  
  RESULTS
  After injury, dry eyes treated with CM-hUCESCs significantly improved epithelial regeneration and showed reduced corneal macrophage inflammatory protein-1 alpha (MIP-1α) and TNF-α mRNA expression as compared to untreated eyes and eyes treated with culture medium or sodium hyaluronate ophthalmic drops. In addition, we found in CM-hUCESCs high levels of proteins, such as tissue inhibitors of metalloproteinases 1 and 2, fibroblast growth factor 6 and 7, urokinase receptor, and hepatocyte growth factor, that could mediate these effects. In vitro, CM-hUCESCs showed a clear bactericidal effect on both E. coli and S. epidermidis and CLs infected with S. epidermidis. Analyses of CM-hUCESCs showed elevated levels of proteins that could be involved in the bactericidal effect, such as the chemokine (C-X-C motif) ligands 1, 6, 8, 10, and the chemokine (C-C motif) ligands 5 and 20.
  
  
  CONCLUSIONS
  Treatment with CM-hUCESCs improved wound healing of alkali-injured corneas and showed a strong bactericidal effect on CLs. Patients using CLs and suffering from dry eye, allergies induced by commercial solutions, or small corneal injuries could benefit from this treatment.
authors.affiliation: Department of Surgery, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Bermudez
authors.affiliation: Department of Physiology, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Sendon-Lago
authors.affiliation: Unidad de Investigación, Fundación Hospital de Jove, Gijón, Spain Fundación para la Investigación con Células Madre Uterinas (FICEMU), Gijón, Spain.
authors.forename: Noemi
authors.initials: N
authors.lastname: Eiro
authors.affiliation: Servicio de Microbiología, Complejo Hospitalario Universitario de Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Treviño
authors.affiliation: Department of Surgery, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), University of Santiago de Compostela, Santiago de Compostela, Spain Service of Ophthalmology, Complejo Hospitalario Universitario de Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Gonzalez
authors.affiliation: Department of Applied Physics (Optometry Area), Optic and Optometry Faculty, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Yebra-Pimentel
authors.affiliation: Department of Applied Physics (Optometry Area), Optic and Optometry Faculty, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Maria Jesus
authors.initials: MJ
authors.lastname: Giraldez
authors.affiliation: Department of Obstetrics and Gynecology, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Macia
authors.affiliation: Unidad de Investigación, Fundación Hospital de Jove, Gijón, Spain.
authors.forename: Maria Luz
authors.initials: ML
authors.lastname: Lamelas
authors.affiliation: Unidad de Investigación, Fundación Hospital de Jove, Gijón, Spain Service of Ophthalmology, Fundación Hospital de Jove, Gijón, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Saa
authors.affiliation: Unidad de Investigación, Fundación Hospital de Jove, Gijón, Spain Fundación para la Investigación con Células Madre Uterinas (FICEMU), Gijón, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Vizoso
authors.affiliation: Department of Physiology, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Roman
authors.initials: R
authors.lastname: Perez-Fernandez
mesh_terms: D000468:Alkalies / Q000633:toxicity; D000818:Animals; D002057:Burns, Chemical / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000166:cytology*; D065306:Corneal Injuries / Q000378:metabolism / Q000382:microbiology / Q000473:pathology*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D019573:Epithelium, Corneal / Q000378:metabolism / Q000382:microbiology / Q000473:pathology*; D005126:Eye Burns / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D015818:Eye Infections, Bacterial / Q000378:metabolism / Q000382:microbiology / Q000473:pathology*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D012313:RNA / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D014945:Wound Healing; D014946:Wound Infection / Q000382:microbiology / Q000473:pathology*
chemical_list: D000468:Alkalies; D017077:Culture Media, Conditioned; D016207:Cytokines; D012313:RNA
keywords: E. coli; S. epiderimidis; corneal ulcers; dry eye; secretoma