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P. D. Lackey, R. Siwale, L. Muya, A. M. Wright; Low Residual Isopropanol Concentration Exposure in Human Corneal Epithelial Cells in vitro. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4618.
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© ARVO (1962-2015); The Authors (2016-present)
Isopropanol (IPA) may be used as an extraction vehicle for polymeric medical devices. Dependent upon processing, low level residual amounts of IPA may remain in the finished device. The purpose of this study was to evaluate the possible inflammatory response of mammalian cells exposed to low residual levels of isopropanol using in vitro cell culture models
HCE-T cells served as the in vitro cell culture model. IPA was tested at 1ppm, 10 ppm and 100 ppm. Exposure time points were 2, 4 and 24 hours. Cellular viability of HCE-T cells was assessed by Alamar BlueTM (AB). IPA concentration in media was determined by GC. Cytokine determination was evaluated by SABioscience and RnD systems inflammatory cytokines kits. NF-kb was determined by Western blot technique. Culture media was the negative control and Benzalkonium chloride (BAC) was the positive control.
HCE-T cells were viable at all IPA test concentrations at all time points. In HCE-T cells challenged with IPA at 100 ppm for 24 hours, IL-8 was slightly upregulated compared to controls. In HCE-T cells challenged with IPA at 1, 10 ppm and 100ppm for 2 or 4 hours, IL-8, IL-6 and NF-kB were not upregulated compared to controls. IL-8, IL-6 or NF-Kb at 1 or 10 ppm at 24 hours was not upregulated in HCE-T cells.
Low residual levels of IPA did not stimulate cytokine release from HCE-T cells in vitro. Continued research will analyze the release of cytokines from a 3D model of human corneal epithelial cells in vitro and PGE2 from macrophages in vitro.
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