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Brittany Coats, Daniel Shedd, Justin Jones; Temporal Changes in Vitreous Inflammatory Cytokines and Neurofilament Heavy Chain Following Ocular Blast Trauma. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6395.
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© ARVO (1962-2015); The Authors (2016-present)
Common closed globe injuries due to blast exposure include corneal swelling and neovascularization, retinal degeneration, and optic neuropathy. Our previous blast rodent model exhibited a temporal multiplexed response in these injuries that may be exploited for drug delivery and protein biomarker detection. The goal of this study was to measure vitreous proteomic changes in inflammatory cytokines and neurofilament heavy chain (NfH) over 8 weeks following blast exposure and correlate findings with the previous injury pathology time-course.
Male Long Evans rats (n = 20) were placed in a 6 meter long by 15.24 cm internal diameter blast tube, and laterally exposed to a 34 psi overpressure blast with a 7 msec duration. Control animals (n=12) were anesthetized and placed in the shock tube, but no pressure wave was administered. Animals were euthanized at 1 day, 1 week, 4 weeks, or 8 weeks post-blast, at which time the eyes were harvested and the vitreous humor extracted and frozen until analysis. Inflammatory cytokines were evaluated using a commercially available kit (RayBio Rat Cytokine Antibody Array G), and evaluated at 532 nm. NfH was measured according to an ELISA technique previously published. The absorbance was read at 450 nm with a reference wavelength of 750 nm. The time-dependent response of all proteins, and comparison of experimental to control levels of concentration at each time point were evaluated with two one-way ANOVAs. A p-value < 0.05 considered significant.
There was an immediate inflammatory cytokine response following the injury with significant increases in CNTF, GMSF, IL-α, IL-β, IL-10, LIX, Leptin, MIP-3, and β-NGF. These increases tended to return to baseline levels at 8 weeks. NfH significantly greater than controls immediately following blast exposure, was sustained for four weeks, and significantly decreased at 8 weeks. All proteomic changes preceded corneal and retinal damage visualized with OCT.
The immediate proteomic changes found in this study were prior to retinal and corneal pathology visualized on OCT, suggesting that cytokines and NfH may be used as biomarkers for damage progression. The profound inflammatory response before visual injury identifies a potential avenue of exploration for drug treatment to mitigate or reduce ocular damage from blast exposure.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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