title: Morphological changes and viability of primary cultured human ocular trabecular meshwork cells after exposure to air.
abstract: OBJECTIVE
  To investigate the possible toxic effect of air exposure for an in vitro model of primary human ocular trabecular meshwork cells (HTM).
  
  
  METHODS
  HTM were isolated from five donor eyes and cultivated at 37 °C. After reaching confluence the cells were seeded on two well chamber slides. The chamber slides were turned upside down in a Petri culture dish full of culture medium and filled with air using a 5 ml syringe, starting this way the exposure of the cells to the air. Subsequently they were placed in the incubation chamber at 37 °C. Six groups of HTM cultures were set up: group 1 consisted of samples in which HTM were exposed to air for 30 min, group 2 for 1 h, group 3 for 3 h, group 4 for 6 h, group 5 for 12 h and group 6 for 24 h.
  
  
  RESULTS
  At 3 h after exposure, the morphology of the cells was still intact, at 6 h few cells appeared deformed and exhibited characteristics of more senescent cells. At 12 h after exposure to air the HTM cells started losing their typical morphology and appeared enlarged and compromised. Viability was superior to 94% in groups 1-3 while for groups 4, 5, 6 it was 82.7%, 39.5% and 12.7% respectively.
  
  
  CONCLUSIONS
  The toxic effect of air exposure for the studied in vitro model of HTM is not significant for the time period of one to three hours. However it starts reducing viability and alternating morphology 6 h after exposure until the time period of 24 h, where the percentage of living cells is drastically decreased. Therefore, we suggest that the use of an air bubble especially in glaucomatous patients should be applied with caution.
authors.affiliation: Moorfields Eye Hospital, London, United Kingdom. Electronic address: Nick.Kopsachilis@moorfields.nhs.uk.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Kopsachilis
authors.affiliation: Department of Ophthalmology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Konstantinos T
authors.initials: KT
authors.lastname: Tsaousis
authors.affiliation: Moorfields Eye Hospital, London, United Kingdom.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Carifi
authors.affiliation: Department of Ophthalmology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Welge-Luessen
mesh_terms: D000388:Air; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000502:physiology*; D006801:Humans; D014129:Trabecular Meshwork / Q000166:cytology*
keywords: Air bubble; Air toxicity; Descemet’s membrane endothelial keratoplasty; Trabecular meshwork cells