title: Effect of Amniotic Membrane Suspension (AMS) and Amniotic Membrane Homogenate (AMH) on Human Corneal Epithelial Cell Viability, Migration and Proliferation In Vitro.
abstract: To analyze the effects of different concentrations of amniotic membrane suspension (AMS) or amniotic membrane homogenate (AMH) on human corneal epithelial cell (HCEC) viability, migration and proliferation.
  
  
  
  Amniotic membranes (AMs) of 13 placentas were prepared and stored at -80°C. For AMS preparation, following de-freezing, AM pieces were inserted in six-well plates and 5 ml Dulbecco's Modified Eagle's Medium (DMEM)/F12 (with 5% fetal bovine serum, FBS) per gram tissue was added for 96 h. After removal of the AM, the remaining supernatant was collected for experiments. For AMH preparation, following de-freezing, AMs were homogenized in liquid nitrogen and 5 ml DMEM/F12 (with 5% FBS) per gram tissue was added. Following centrifugation, the supernatant was collected for experiments. HCECs were expanded and incubated in DMEM/F12, 5% FBS supplemented by 15%, 30% or 100% AMS or 15% or 30% AMH. Viability was analyzed using Cell Proliferation Kit XTT, migration using wound healing assay and proliferation by the cell proliferation ELISA BrdU kit.
  
  
  
  HCEC viability remained unchanged using 15% or 30% AMS (p = 1.0 for both); however, it decreased significantly using 100% AMS (p < 0.001) or 15% (p = 0.041) or 30% AMH (p < 0.001), compared to controls. Using 15% or 30% AMS, HCEC migration increased significantly (p < 0.001 for both). Using 15% or 30% AMH (p = 0.153; p = 0.083), HCEC migration remained unchanged and 100% AMS inhibited HCEC migration (p < 0.001). Next, 15% and 30% AMS had no effect on HCEC proliferation (p = 0.454 and p = 0.119), but 100% AMS (p < 0.001) and 15% (p = 0.002) and 30% AMH (p = 0.001) inhibited HCEC proliferation significantly.
  
  
  
  With unchanged HCEC viability and proliferation and increased HCEC migration, 15% and 30% AMS application seems to be the most appropriate method to support epithelial healing.
authors.affiliation: a Department of Ophthalmology , Saarland University Medical Center , Homburg , Saar , Germany.
authors.forename: Ming-Feng
authors.initials: MF
authors.lastname: Wu
authors.affiliation: a Department of Ophthalmology , Saarland University Medical Center , Homburg , Saar , Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Stachon
authors.affiliation: b Experimental Ophthalmology , Saarland University , Homburg , Saar , Germany.
authors.forename: Achim
authors.initials: A
authors.lastname: Langenbucher
authors.affiliation: a Department of Ophthalmology , Saarland University Medical Center , Homburg , Saar , Germany.
authors.forename: Berthold
authors.initials: B
authors.lastname: Seitz
authors.affiliation: a Department of Ophthalmology , Saarland University Medical Center , Homburg , Saar , Germany.|c Department of Ophthalmology , Semmelweis University , Budapest , Hungary.
authors.forename: Nóra
authors.initials: N
authors.lastname: Szentmáry
mesh_terms: D000328:Adult; D000650:Amnion; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D065306:Corneal Injuries / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D019573:Epithelium, Corneal; D005260:Female; D006801:Humans; D011247:Pregnancy; D055815:Young Adult
chemical_list: D017077:Culture Media, Conditioned
keywords: Amniotic membrane; amniotic membrane homogenate; amniotic membrane suspension; human corneal epithelial cells; migration; proliferation; viability