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Veeral S. Shah, Thomas Friberg, James L. Funderburgh, Danny S. Roh, Daewon Park, Yadong Wang; Analysis of Cellular Toxicity of a Thermal Gel for utilization as a Retinal Drug Delivery System. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6463.
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Reverse thermal gels are chemicals that readily dissolve in water and undergo phase transition to a hydrogel with changes in ambient temperature. Thermal gels can 1) be designed to undergo gelation at body temperature and 2) be modified to incorporate bio-molecules to guide cell interaction. Thus they have numerous biomedical implications including tissue engineering and drug delivery. In this study, we tested the toxic effects of a novel thermal gel Polyurethane/ Poly-ethylene-glycol (ESHU) on cultured bovine corneal endothelium cells by evaluating cell viability.
Cultured monolayers of primary bovine corneal endothelium were exposed for 24 hours to one of the following 4 liquids: 1) Control (serum-free DMEM), 2) ESHU (15% dilution of DMEM), 3) perfluoron (PFO or perfluoro-n-octane) and 4) a 5000cs silicone oil. After at 1 hr, 12 hr, and 24 hrs viability of cultured corneal endothelium cells was evaluated by immunofluorescent microscopy after staining with Calcein AM, propidium iodide (PI), and Hoechst. Cytotoxicity was calculated as PI nuclei/total nuclei in three separate cultures. Cell morphology was also assessed by microscopy.
Cytotoxicity analysis demonstrated that there was no significant damage to cultured corneal endothelium cells by ESHU gel or PFO treatment compared to the control (P>0.05, two- way ANOVA) at each timepoint. Silicone oil treated cultures, however, demonstrated marked cell death at all timepoints when compared to controls (P<0.001, two- way ANOVA) and ESHU gel treatment cultures (P<0.001, two- way ANOVA), with nearly 75% cell death at 24 hours. Qualitatively, corneal endothelium cells treated with ESHU thermal gel for 48 hr showed no evidence of altered cellular morphology on microscopic examination.
In this cell culture model, ESHU thermal gel appear safe with respect to limited contact with bovine corneal endothelium, and support a practical role for further investigations of thermal gels in drug delivery.
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