Effects of Perfluorobutylpentane (F4H5) on Corneal Endothelial Cells

Purpose: To evaluate the effects of perfluorobutylpentane (F4H5) on corneal endothelial cell density (ECD) and morphology using a porcine corneal endothelial organ culture model. Materials and methods: "Split corneal buttons" were cultivated for 15 days (d) after incubation in F4H5 (15, 30, 60, and 120 min) or BSS (controls). ECD was assessed manually on d1, d8, and d15. After histological staining (trypan blue, alizarin red S) on d15 morphological changes (reformation figures, rosette formations, and alizarin red cells) were evaluated. Results: ECD was significantly reduced after incubation in F4H5 for 120 min (median ± 25%/75%-quartile; 3281 ± 43/222 cells/mm²; p = 0.046) on d15 compared to controls (3658 ± 129/296 cells/mm²), but not after shorter incubation times (15, 30, and 60 min). Morphological assessment supports these findings as reformation figures (F4H5 120 min: 10.5 ± 9.3/13.9/mm² vs. controls: 5.2 ± 2.8/7.2/mm²; p = 0.010), rosette formations (F4H5 120 min 25.566 ± 17.044/36.219/mm² vs. controls: 8.333 ± 0.000/15.667/mm²; p = 0.002), and alizarin red cells (F4H5 120 min: 38.350 ± 29.827/51.333/mm² vs. controls: 20.833 ± 10.417/25.000/mm²; p = 0.049) were significantly more prevalent after incubation in F4H5 for 120 min compared to controls. Also, F4H5 60 min showed significantly more rosette formations (25.452 ± 16.968/36.057/mm²; p = 0.006) and alizarin red cells (46.662 ± 42.420/50.903/mm²; p = 0.007), but not reformation figures (7.0 ± 2.2/1.6 %; p = 0.953). Conclusion: Short exposure (≤30 min) of porcine corneal endothelial cells to F4H5 does not have significant effects on ECD or morphological characteristics. Longer exposure times (≥60-120 min) may cause ECD decline and/or induce morphological changes.