title: Determination of a No-Observable Effect Level for Endotoxin Following a Single Intravitreal Administration to Dutch Belted Rabbits.
abstract: The purpose of this study was to characterize the inflammatory response and determine a no-observable effect level (NOEL) in rabbit eyes after endotoxin intravitreal (ITV) injection.
  
  
  
  Fifty-three naïve male Dutch Belted rabbits were treated with a single 50-μL ITV injection ranging from 0.01 to 0.75 endotoxin units/eye (EU/eye) and monitored for up to 42 days post treatment. Ophthalmic examination included slit-lamp biomicroscopy and indirect ophthalmoscopy. Laser flare photometry was performed in a subset of animals. On days 2, 8, 16, and 43, a subset of animals was necropsied and eyes processed for histopathological evaluation.
  
  
  
  Intravitreal injection of endotoxin at ≥0.05 EU/eye resulted in a dose-related anterior segment inflammation response. No aqueous flare or cell response was noted in the 0.01 EU/eye dose group. A more delayed posterior segment response characterized by vitreal cell response was observed beginning on day 5, peaking on day 9, and decreasing starting on day 16 that persisted at trace to a level of 1+ on day 43. Microscopy findings of infiltrates of minimal mixed inflammatory cells in the vitreous and subconjunctiva and proteinaceous fluid in the anterior chamber and/or vitreous were observed in eyes given ≥0.1 EU/eye.
  
  
  
  We defined the NOEL for ITV endotoxin to be 0.01 EU/eye, suggesting that the vitreal cavity is more sensitive to the effects of endotoxin than the anterior segment and aqueous chamber. These data highlight the importance of assessing endotoxin level in intravitreal formulations, as levels as low as 0.05 EU/eye may confound the safety evaluations of intravitreal therapeutics in rabbits.
authors.affiliation: Department of Safety Assessment, Genentech, Inc., South San Francisco, California, United States.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Bantseev
authors.affiliation: Department of Surgical Services, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Miller
authors.affiliation: Department of Surgical Services, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States.
authors.forename: Ellison
authors.initials: E
authors.lastname: Bentley
authors.affiliation: Department of Safety Assessment, Genentech, Inc., South San Francisco, California, United States.
authors.forename: Chris
authors.initials: C
authors.lastname: Schuetz
authors.affiliation: Covance Laboratories, Inc., Madison, Wisconsin, United States.
authors.forename: Tim M
authors.initials: TM
authors.lastname: Streit
authors.affiliation: Covance Laboratories, Inc., Madison, Wisconsin, United States.
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Christian
authors.affiliation: Farman Pathology, Reno, Nevada, United States.
authors.forename: Cindy
authors.initials: C
authors.lastname: Farman
authors.affiliation: Department of Safety Assessment, Genentech, Inc., South San Francisco, California, United States.
authors.forename: Helen
authors.initials: H
authors.lastname: Booler
authors.affiliation: Department of Safety Assessment, Genentech, Inc., South San Francisco, California, United States.
authors.forename: Evan A
authors.initials: EA
authors.lastname: Thackaberry
mesh_terms: D000818:Animals; D000869:Anterior Eye Segment / Q000187:drug effects* / Q000473:pathology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004596:Electroretinography; D004731:Endotoxins / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000633:toxicity*; D058449:Intravitreal Injections; D008297:Male; D009887:Ophthalmoscopy; D010783:Photometry; D011817:Rabbits; D012160:Retina / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D014606:Uveitis, Anterior / Q000139:chemically induced* / Q000175:diagnosis / Q000378:metabolism
chemical_list: D004731:Endotoxins