title: Cytotoxicity and genotoxicity of intravitreal adalimumab administration in rabbit retinal cells.
abstract: OBJECTIVE
  To assess the cytotoxicity and genotoxicity of intravitreal adalimumab treatment in an animal experimental model using cytological and molecular techniques.
  
  
  METHODS
  Eighteen rabbits were randomly assigned to three groups: control, adalimumab treatment, and placebo. Cytotoxicity on retinal cells was evaluated using flow cytometry assays to determine the level of apoptosis and necrosis. Genotoxicity was evaluated by comet assays to assess DNA damage, and quantitative real-time polymerase chain reaction (qPCR) was used to evaluate expression of apoptosis-inducing caspases (8 and 3).
  
  
  RESULTS
  No cytotoxicity or genotoxicity was observed in any of the two treatment groups (adalimumab and placebo) following intravitreal administration compared with the control group. Flow cytometry analysis revealed that more than 90% of the cells were viable, and only a low proportion of retinal cells presented apoptotic (~10%) or necrotic (<1%) activity across all groups. Molecular damage was also low with a maximum of 6.4% DNA degradation observed in the comet assays. In addition, no increase in gene expression of apoptosis-inducing caspases was observed on retinal cells by qPCR in both the adalimumab and placebo groups compared with the control group.
  
  
  CONCLUSIONS
  The use of adalimumab resulted in no detectable cytotoxicity or genotoxicity on retinal cells for up to 60 days upon administration. These results therefore indicate that adalimumab may be a safe option for intravitreal application to treat ocular inflammatory diseases in which TNF-α is involved.
authors.affiliation: Ophthalmic Center Reference, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Álcio Coutinho de
authors.initials: ÁC
authors.lastname: Paula
authors.affiliation: Ophthalmic Center Reference, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Marcos Pereira de
authors.initials: MP
authors.lastname: Ávila
authors.affiliation: Ophthalmic Center Reference, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: David Leonardo Cruvinal
authors.initials: DL
authors.lastname: Isaac
authors.affiliation: Ophthalmic Center Reference, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Salustiano
authors.affiliation: Molecular Genetics and Cytogenetics, Institute of Biological Science, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Aliny Pereira de
authors.initials: AP
authors.lastname: Lima
authors.affiliation: Molecular Genetics and Cytogenetics, Institute of Biological Science, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Francyelli Mariana
authors.initials: FM
authors.lastname: Mello
authors.affiliation: Molecular Genetics and Cytogenetics, Institute of Biological Science, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Flávia de Castro
authors.initials: Fde C
authors.lastname: Pereira
authors.affiliation: Instituto de Ciências Farmacêuticas Estudos e Pesquisa, Goiânia, GO, Brazil.
authors.forename: Pedro Henrique de Paula
authors.initials: PH
authors.lastname: Silva
authors.affiliation: Molecular Genetics and Cytogenetics, Institute of Biological Science, Federal University of Goiás, Goiânia, GO, Brazil.
authors.forename: Elisângela de Paula Silveira
authors.initials: Ede P
authors.lastname: Lacerda
mesh_terms: D000068879:Adalimumab; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000633:toxicity*; D061067:Antibodies, Monoclonal, Humanized / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000032:analysis / Q000187:drug effects; D053181:Caspase 8 / Q000032:analysis / Q000187:drug effects; D002470:Cell Survival; D020552:Comet Assay; D004249:DNA Damage; D005434:Flow Cytometry; D058449:Intravitreal Injections / Q000379:methods*; D008297:Male; D023421:Models, Animal; D009336:Necrosis; D011817:Rabbits; D011897:Random Allocation; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012160:Retina / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects
chemical_list: D000893:Anti-Inflammatory Agents; D061067:Antibodies, Monoclonal, Humanized; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3; D053181:Caspase 8; D000068879:Adalimumab