title: Inhibition of human retinal pigment epithelial cell attachment, spreading, and migration by alkylphosphocholines.
abstract: OBJECTIVE
  To investigate the effect of alkylphosphocholines (APCs) on human retinal pigment epithelium (RPE) attachment, spreading, migration, and microfilament assembly in vitro.
  
  
  METHODS
  Cultured RPE cells of five human donors were treated with one of four APCs (C18:1-PC, C20:1-PC, C21:1-PC, or C22:1-PC) in the presence of fetal calf serum. Cell viability was tested by the trypan blue exclusion assay. Attachment was assessed after a 2-hour incubation of RPE cells on coated 96-well-plates and subsequent MTT testing. Cellular spreading is characterized by cytoplasmic halo formation and was quantified by counting four separate fields of RPE cells allowed to spread on coated 24-well plates for 4 hours. Migration was measured by a modification of the Boyden chamber method in microchemotaxis chambers with polycarbonated filters. Microfilament assembly was assessed by immunofluorescence analysis after incubation with rhodamine-phalloidin.
  
  
  RESULTS
  All four APCs inhibited RPE cell attachment by more than 70% of their IC50 (C18:1-PC: 30 microM; C20:1-PC: 10 microM; C21:1-PC: 10 microM; and C22:1-PC: 10 microM). Also, APCs inhibited RPE cell spreading by more than 80% and migration by more than 90% at similar concentrations. Trypan blue staining revealed a toxicity within control limits within the concentration interval tested. Microfilament organization was significantly disturbed after incubation of RPE cells with one of the four APCs close to its IC50.
  
  
  CONCLUSIONS
  APCs inhibit RPE cell attachment and spreading in vitro at nontoxic concentrations. As a possible mechanism of action, APCs disturb microfilament assembly, since they are known to interfere with protein kinase C (PKC) function. This could represent a novel method of preventing even early stages of proliferative vitreoretinal diseases like proliferative vitreoretinopathy (PVR).
authors.affiliation: Department of Ophthalmology, Ludwig-Maximilians-University, Munich, Germany.
authors.forename: Kirsten H
authors.initials: KH
authors.lastname: Eibl
authors.forename: Daniel
authors.initials: D
authors.lastname: Kook
authors.forename: Siegfried
authors.initials: S
authors.lastname: Priglinger
authors.forename: Christos
authors.initials: C
authors.lastname: Haritoglou
authors.forename: Alice
authors.initials: A
authors.lastname: Yu
authors.forename: Anselm
authors.initials: A
authors.lastname: Kampik
authors.forename: Ulrich
authors.initials: U
authors.lastname: Welge-Lussen
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000328:Adult; D000368:Aged; D002448:Cell Adhesion / Q000187:drug effects*; D018929:Cell Culture Techniques; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism
chemical_list: D010767:Phosphorylcholine