Cytotoxicity and genotoxicity of intravitreal adalimumab administration in rabbit retinal cells

Abstract

Purpose: To assess the cytotoxicity and genotoxicity of intravitreal adalimumab treatment in an animal experimental model using cytological and molecular te chniques. 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. Conclusion: The use of adalimumab resulted in no detectable cytotoxicity or ge notoxicity 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.

A review of ocular adverse events of biological anti-TNF drugs

“…It has 12 approved indications, including noninfectious intermediate, posterior, and panuveitis. To date, treatment with adalimumab for uveitis is still systemic; nevertheless, studies have been made in the direction of producing a more local therapy using intravitreal injections [86][87][88][89][90]. Although most studies demonstrated safety, it is still a controversial topic, and further research is needed to make this method feasible in the future.…”

Evaluation of the Influence of Adalimumab on the Expression Profile of Leptin-Related Genes and Proteins in Keratinocytes Treated with Lipopolysaccharide A

“…The changes in the cell culture model were noted by these scientists as early as 1 h after the incubation of the cells with the drug [37], which is consistent with our observations. On the other hand, Paula et al did not show a statistically significant difference in the expression of caspases-8 and -9 mRNA, when adalimumab was added to the animal experimental model of retinal cells in comparison with the placebo and the control [38]. The differences between the results presented by Paula et al [38] and our own may result from either using different study models or from the fact that the retinal cells prior to exposition to adalimumab were not stimulated with a proinflammatory factor.…”

Ocular toxicity of intravitreal golimumab in a rabbit model

“…Several studies in the literature have evaluated the apoptotic effect of anti-TNF drugs by immunohistochemistry. Paula et al compared the apoptotic effect of IVT adalimumab, sham injection, and no injection to the rabbits' eyes and found that the caspase gene expression was similar between three groups [37]. Ueda et al found that infliximab and adalimumab increased the amount of active caspase-3 compared to golimumab, thus leading to more apoptotic DNA fragmentation [38] There was no significant difference in the percentage of TUNEL and caspase-3 positive cells compared to the control group in the golimumab group's GCL.…”