Evaluation of pigment epithelium-derived factor concentration in equine amniotic membrane homogenate and its in-vitro vascular endothelial growth factor inhibition effect in tears of dogs with vascularized ulcerative keratitis.

Background: Corneal neovascularization can result from many pathological processes affecting the ocular surface leading to disturbances and opacifications that reduce corneal clarity and may impact vision. In veterinary medicine, the use of topical corticosteroid is contraindicated in the presence of ulcerative keratitis, and there is sparse research regarding safe medical alternatives to inhibit corneal neovascularization in dogs to improve visual outcome. Aim: To investigate the pigment epithelium-derived factor (PEDF) concentration in equine amniotic membrane homogenate (EAMH) and its in-vitro vascular endothelial growth factor (VEGF) inhibition in tears of dogs with vascularized ulcerative keratitis. Methods: Homogenates from 10 equine amniotic membranes (AM) were analyzed by sandwich enzyme-linked immunosorbent assay (ELISA) for quantification of equine PEDF and VEGF. Forty tear samples were collected from both eyes of dogs diagnosed with vascularized ulcerative keratitis, and 50 samples from healthy dogs. Samples from affected eyes were allocated to G1 - affected undiluted tears; G2 - affected tears diluted with phosphate-buffer solution; G3 - affected tears treated with low-concentrated EAMH; and G4 - affected tears treated with high-concentrated EAMH. Tears from the unaffected contralateral eyes were composed in G5, while G6 was composed by tears from healthy dogs (control). The presence and levels of VEGF were evaluated in all groups by Western blot and ELISA. Results: The PEDF:VEGF ratio in EAMH was 110:1. An increase in VEGF levels was observed in tears from eyes with vascularized corneal ulcers (G1) as well as in contralateral tears (G5), compared to normal dogs (G6). High-concentrated EAMH provided a greater decrease in VEGF levels in-vitro compared to low-concentrated EAMH. Conclusion: EAMHs exhibited high concentrations of PEDF in comparison to VEGF and were able to partially decrease VEGF levels in tears of dogs with vascularized ulcers, in-vitro. Our results suggest that VEGF concentration is elevated in tears of dogs with active vascularized ulcerative keratitis in both affected and contralateral eyes compared to that of healthy dogs.

Tear production, intraocular pressure, and central corneal thickness in white-tailed deer (Odocoileus virginianus).

OBJECTIVE: To evaluate the tear production, intraocular pressure, and central corneal thickness in healthy white-tailed deer (Odocoileus virginianus) and assess for seasonal variability. ANIMALS STUDIED: Nineteen healthy fawns (3-10 days old) and 8 healthy adult white-tailed deer (164-280 days old) were studied. PROCEDURES: All fawns were manually restrained for the study, while adult animals were immobilized with an intramuscular injection of combined xylazine (2.2 mg/kg), tiletamine (1 mg/kg), and zolazepam (1 mg/kg). Tear production (STT-1) was evaluated in both eyes of all fawns. Intraocular pressure (applanation and rebound tonometry) and central corneal thickness (ultrasound pachymetry) were determined in both eyes of all fawns and adult deer. RESULTS: Mean ± SD tear production in fawns was 12.32 ± 4.46 mm/min. Mean ± SD IOP values in the fawns were 16.21 ± 4.97 mm Hg (TonoPen® ) and 14.05 ± 5.03 mm Hg (TonoVet® ). In adult deer, mean ± SD IOP values were 15.57 ± 2.88 mm Hg with TonoPen® and 12.87 ± 2.57 with TonoVet® , and TonoPen® significantly overestimated the IOP, compared to TonoVet® (P < 0.005). The IOP was not significantly different between young and adult animals (P = 0.22). A significant difference (P < 0.005) in CCT values was observed between fawns (528.42 ± 27.65 µm) and adult deer (747.55 ± 41.60 µm). CONCLUSIONS: This study reports normal values of tear production in healthy fawns, and intraocular pressure and central corneal thickness in healthy young and adult white-tailed deer.