Effect of sorafenib in a murine high risk penetrating keratoplasty model

AIM： To evaluate the effect of sorafenib in murine high risk keratoplasty model. ~ METHODS： Graft survival, corneal neovascularization, and corneal lymphangiogenesis were compared among the sorafenib, dexamethasone, dimethyl sulfoxide （DMSO）, and phosphate buffered saline （PBS） groups following subconjunctival injection in mice that underwent high risk penetrating keratoplasty （HRPK）. Real-time polymerase chain reaction was performed to quantify the expression of inflammatory cytokines and vascular endothelial growth factor （VEGF）-A, VEGF-C, vascular endothelial growth factor receptor （VEGFR）-2, VEGFR-3. RESULTS： The two-month graft survival rate for HRPK was 42.86% in sorafenib group, 37.50% in dexamethasone group, 0 in DMSO group, and 0 in PBS group. Sorafenib significantly increased graft survival compared to the DMSO and PBS group （P〈0.05）. The sorafenib didn＇t show significant effect in decreasing neovascularization compared with dexamethsone, DMSO, and PBS group. The sorafenib showed less total lymphangiogenesis than the dexamethasone, DMSO, and PBS group （P=0.011, P〈0.001, P〈0.001, respectively）. The sorafenib group showed reduced expression of VEGF-C, tumor necrosis factor （TNF）-alpha, interleukin （IL）-6, VEGFR-2 and VEGFR-3 compared with DMSO group and PBS group （all P〈0.05）. The sorafenib group didn＇t show difference in the expression of VEGF-A compared with DMSO, neither withPBS. The sorafenib group showed reduced expression of VEGFR-3 compared with dexamethasone （/〉=-0.051）. CONCLUSION： The subconjunctivally administered sorafenib shows significant anti-lymphangiogenic effect, resulting in increased transplant survival in a murine high risk keratoplasty model. We suggest that a close linkage between decreased VEGF-C/VEGFR-2 and -3 signaling and increased corneal graft survival by sorafenib seems to exist.

Effect of trapping vascular endothelial growth factor-A in a murine model of dry eye with inflammatory neovascularization

AIM： To evaluate whether trapping vascular endothelial growth factor-A（VEGF-A） would suppress angiogenesis and inflammation in dry eye corneas in a murine corneal suture model.METHODS： We established two groups of animals, one with non-dry eyes and the other with induced dry eyes.In both groups, a corneal suture model was used to induce inflammation and neovascularization. Each of two groups was again divided into three subgroups according to the treatment; subgroup I（aflibercept）,subgroup II（dexamethasone） and subgroup III（phosphate buffered saline, PBS）. Corneas were harvested and immunohistochemical staining was performed to compare the extents of neovascularization and CD11b＋ cell infiltration. Real-time polymerase chain reaction was performed to quantify the expression of inflammatory cytokines and VEGF-A in the corneas.RESULTS： Trapping VEGF-A with aflibercept resulted in significantly decreased angiogenesis and inflammation compared with the dexamethasone and PBS treatments in the dry eye corneas（all P 〈0.05）, but with no such effects in non-dry eyes. The anti-inflammatory and antiangiogenic effects of VEGF-A trapping were stronger than those of dexamethasone in both dry eye and non-dry eye corneas（all P 〈0.05）. The levels of RNA expression of VEGF-A, TNF-alpha, and IL-6 in the aflibercept subgroup were significantly decreasedcompared with those in the PBS subgroup in the dry eye group.CONCLUSION： Compared with non-dry eye corneas,dry eye corneas have greater amounts of inflammation and neovascularization and also have a more robust response to anti-inflammatory and anti-angiogenic agents after ocular surface surgery. Trapping VEGF-A is effective in decreasing both angiogenesis and inflammation in dry eye corneas after ocular surface surgery.

Effect of itraconazole on the cornea in a murine suture model and penetrating keratoplasty model

AIM： To investigate the anti-（lymph）angiogenic and/or anti-inflammatory effect of itraconazole in a corneal suture model and penetrating keratoplasty（PK） model. METHODS： Graft survival, corneal neovascularization, and corneal lymphangiogenesis were compared among itraconazole, amphotericin B, dexamethasone, phosphate buffered saline（PBS） and surgery-only groups following subconjunctival injection in mice that underwent PK and corneal suture. Immunohistochemical staining and analysis were performed in each group. Real-time polymerase chain reaction（RT-PCR） was performed to quantify the expression of inflammatory cytokines（TNF-alpha, IL-6） and vascular endothelial growth factor（VEGF）-A, VEGF-C, VEGFR-2, and VEGFR-3.RESULTS： In the suture model, the itraconazole group showed less angiogenesis, less lymphangiogenesis, and less inflammatory infiltration than the PBS group（all P〈0.05）. The itraconazole group showed reduced expression of VEGF-A, VEGFR-2, TNF-alpha, IL-6 than the PBS group（all P〈0.05）. In PK model, the two-month graft survival rate was 28.57% in itraconazole group, 62.50% in dexamethasone group, 12.50% in PBS group, 0 in amphotericin B group and 0 in surgery-only group. Graft survival in the itraconazole group was higher than that in the amphotericin, PBS and surgery-only group（P=0.057, 0.096, 0.012, respectively）. The itraconazole group showed less total angiogenesis and lymphangiogenesis than PBS group（all P〈0.05）.CONCLUSION： Itraconazole decrease neovascularization, lymphangiogenesis, and inflammation in both a corneal suture model and PK model. Itraconazole has anti-（lymph）-angiogenic and anti-inflammatory effects in addition to its intrinsic antifungal effect and is therefore an alternative treatment option in cases where steroids cannot be used.