Soluble vascular endothelial growth factor receptor 3 is essential for corneal alymphaticity.

Corneal transparency is a prerequisite for optimal vision and in turn relies on an absence of blood and lymphatic vessels, which is remarkable given the cornea's proximity to vascularized tissues. Membrane-bound vascular endothelial growth factor receptor 3 (VEGFR-3), with its cognate ligand vascular endothelial growth factor C (VEGF-C), is a major mediator of lymphangiogenesis. Here, we demonstrate that the cornea expresses a novel truncated isoform of this molecule, soluble VEGFR-3 (sVEGFR-3), which is critical for corneal alymphaticity, by sequestering VEGF-C. sVEGFR-3 binds and sequesters VEGF-C, thereby blocking signaling through VEGFR-3 and suppressing lymphangiogenesis induced by VEGF-C. sVEGFR-3 knockdown leads to lymphangiogenesis and hemangiogenesis in the mouse cornea, while overexpression of sVEGFR-3 inhibits lymphangiogenesis and hemangiogenesis in a murine suture injury model. Pax6(+/-) mice spontaneously develop corneal and lymphatic vessels and are deficient in sVEGFR-3. sVEGFR-3 suppresses hemangiogenesis by blocking VEGF-C-induced phosphorylation of VEGFR-2. Overexpression of sVEGFR-3 leads to a 5-fold increase in corneal transplant survival in mouse models. sVEGFR-3 holds promise as a molecule to control and regress lymphatic-vessel-based dysfunction. Therefore, sVEGFR-3 has the potential to protect the injured cornea from opacification secondary to infection, inflammation, or transplant rejection.

In Vivo Human Lacrimal Gland Imaging Using an Ultrasound Biomicroscopy.

PURPOSE: In the present study, we introduce human lacrimal gland imaging using an ultrasound biomicroscopy (UBM) with a soft cover and show their findings. METHODS: The representative UBM findings of palpebral lobes in seven subjects (four with non-Sjögren dry eye syndrome, one with Sjögren syndrome, and two healthy subjects) were described in this study. To prolapse the palpebral lobe, the examiner pulled the temporal part of the upper eyelid in the superotemporal direction and directed the subject to look in the inferonasal direction. We scanned the palpebral lobes longitudinally and transversely using UBM. We used an Aviso UBM with a 50 MHz linear probe and ClearScan. RESULTS: In UBM of two healthy subjects, the echogenicity of the lacrimal gland was lower than that of the sclera and homogeneous. But the parenchyma of a patient with Sjögren dry eye syndrome was quite inhomogeneous compared to the healthy subjects. In two patients with dry eye syndrome, we were able to observe some lobules in the parenchyma. We could find excretory ducts running parallel at the surface of the longitudinal section in some subjects. In the longitudinal UBM scan of a subject, we observed a tubular structure at a depth of 1,500 µm that was considered a blood vessel. It ran from the superonasal to the inferotemporal direction. In a subject, we observed a large cyst beneath the conjunctiva. CONCLUSIONS: Lacrimal gland imaging using UBM has both advantages of optical coherence tomography and sonography, and could be useful for evaluating dry eye syndrome.

Stability of neodymium:YAG laser posterior capsulotomy in eyes with capsular tension rings.

AIM: To evaluate the stability of neodymium (Nd):YAG laser posterior capsulotomy in eyes with capsular tension rings (CTRs). METHODS: A total of 60 eyes that underwent cataract surgery and laser posterior capsulotomy postoperatively were included in this retrospective cohort study. To evaluate the safety and stability of capsulotomy, changes in the size of posterior capsulotomy and anterior chamber depth (ACD) in three groups: the group without CTR, the group with 12 mm CTRs, and the group with 13 mm CTRs, at 1wk, 3, 12, and 15mo after capsulotomy, were compared. RESULTS: In the group without CTR and the group with 12 mm CTR, there was no significant change in ACD at every post-laser follow-up. In the group with 13 mm CTR, the ACD change was significant until 3mo after capsulotomy. In all groups, there was a significant increase in the area of capsulotomy between 1wk and 3mo post-laser. Between 3 and 12mo post-laser, only the group with 13 mm CTR showed a significant increase in the area of capsulotomy (P<0.01). CONCLUSION: Laser posterior capsulotomy is safe in all three groups. The capsulotomy and ACD become stabilized and have not shown significant changes since 1y post-laser, even with larger CTRs. The maintenance of centrifugal capsular tension can last longer with larger CTRs, and the stability of the capsulotomy site can be reached about 12mo after capsulotomy in pseudophakic eyes with larger CTRs.

Efficacy of haptic sutured in-the-bag intraocular lens for intraocular lens-capsule complex stability: a comparison of three insertion methods.

AIM: To evaluate the efficacy and stability of haptic sutured in-the-bag intraocular lens (IOL) in eyes with zonular instability. METHODS: A total 60 eyes of 60 patients were included in this retrospective cohort study. Postoperative stability in three groups [haptic sutured IOL in the bag, IOL in the bag insertion with haptics oriented toward areas of zonulysis, IOL with capsular tension ring (CTR) in the bag insertion] were compared according to the IOL insertion methods. To evaluate the IOL stability, the changes of anterior chamber depth (ACD), refraction, contraction of anterior continuous curvilinear capsulotomy (CCC) area, and tilt of IOL were compared. RESULTS: There was no significant difference in change of ACD (-0.04±0.01 mm in group of haptic sutured IOL, -0.07±0.01 mm in group of CTR insertion) and refraction (0.05±0.05 D in group of haptic sutured IOL, 0.37±015 D in group of CTR insertion) between the group of haptic sutured IOL in the bag and CTR insertion group. But in comparison of CCC contraction and IOL tilt, CTR insertion group showed less contraction (1.00%±0.52%) and less IOL tilt (2.66°±0.11°) than the group of haptic sutured IOL in the bag (6.32%±1.36%, 3.47°±0.11°, respectively). The CTR insertion group showed the least CCC contraction and the least tilt. CONCLUSION: In eyes with zonular instability, the method of haptic sutured IOL in-the-bag shows comparable stability in ACD and refraction in comparison with IOL with CTR in the bag insertion. The method of IOL only in-the-bag insertion shows the largest contraction of CCC and the largest tilt of IOL.

Aqueous humor analyses of diabetic macular edema patients with subretinal fluid.

We identified treatment-naïve diabetic macular edema (DME) patients with or without subretinal fluid (SRF). We compared their baseline characteristics: aqueous concentrations of interleukin (IL)-1ß, IL-2, IL-6, IL-8, IL-10, and IL-17, as well as tumor necrosis factor-α, vascular endothelial growth factor (VEGF), and placental growth factor (PlGF). We also compared fundus and optical coherence tomography (OCT) findings, and responsiveness to anti-VEGF treatments. Of 67 DME patients, 18 (26.87%) had SRF. Compared to the no SRF group, the SRF group had significantly higher levels of IL-6, IL-8, VEGF, and PlGF in aqueous humor. After grouping according to diabetic retinopathy stage, non-proliferative diabetic retinopathy (NPDR) patients with SRF had higher aqueous levels of IL-6 and IL-8, compared to NPDR patients without SRF. Moreover, proliferative diabetic retinopathy (PDR) patients with SRF had higher aqueous levels of VEGF and PlGF, compared to PDR patients without SRF. Fundus and OCT analyses revealed that the SRF group had a greater proportion of patients with succinate or patch-shaped hard exudates involving the macula, and greater central subfield thickness (CST) at baseline. After 6 months of anti-VEGF treatments, the SRF group showed better responsiveness in terms of CST; however, visual acuity was not correlated with responsiveness. Considering higher aqueous levels of VEGFs and pro-inflammatory cytokines, SRF could be a biomarker related to diabetic retinopathy activity. DME patients with SRF showed better anatomical responsiveness to anti-VEGF treatments, but did not show better functional improvement on short-term evaluation compared to those of DME patients without SRF.

Maxillary Zoster and Neurotrophic Keratitis following Trigeminal Block.

Herpes zoster ophthalmicus is commonly used to describe viral reactivation from the trigeminal ganglia with ocular involvement. The ophthalmic branch is the most commonly involved, whereas the maxillary and mandibular dermatomes are less commonly affected. Neurotrophic ulcer may occur secondary to intentional or inadvertent damage to the trigeminal nucleus, root, ganglion, or any segment of the ophthalmic branch of this cranial nerve. We report a case of reactivated maxillary herpes zoster combined with neurotrophic keratitis due to percutaneous 2nd and 3rd branch of trigeminal nerve block with alcohol to treat trigeminal neuralgia. A 57-year-old female came to the ophthalmology department complaining of decreased visual acuity and skin vesicle over the right lower lid and cheek. She had undergone right trigeminal nerve block for treatment of trigeminal neuralgia. Clinical examination revealed neurotrophic keratitis and maxillary herpes zoster. She was treated with oral and topical antivirals and vigorous lubrication with eye drops. Her neurotrophic keratitis showed a slow recovery. Although a few cases of herpes zoster following nerve block have been described, it would appear that a case of simultaneous maxillary herpes zoster and neurotrophic keratitis following trigeminal block has not yet been documented. It is possible that trigeminal nerve block may cause reactivation of latent virus and refractory neurotrophic keratitis.

Antiangiogenesis Effect of Albendazole on the Cornea.

Purpose: To investigate the anti-(lymph)angiogenic and anti-inflammatory effects of albendazole and to study whether these effects are additive with bevacizumab therapy in a murine corneal suture model. Methods: Corneal neovascularization (NV) and lymphangiogenesis (LY) were compared in a corneal suture model after administration of a subconjunctival injection of albendazole, bevacizumab, dexamethasone, or phosphate-buffered saline (PBS). 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 (tumor necrosis factor [TNF]-alpha and interleukin-6), vascular endothelial growth factor (VEGF)-A, VEGF-C, vascular endothelial growth factor receptor (VEGFR)-2, and VEGFR-3. To evaluate the additive effect of albendazole, corneal NV and LY were also analyzed in a combined group of albendazole and bevacizumab therapy and the additive effect was compared with that in the group of double dose of bevacizumab. Results: The albendazole group showed less NV and less LY compared with the PBS control group (P < 0.01). When albendazole was combined with bevacizumab therapy, a significant decrease in NV and LY was seen compared with bevacizumab treatment alone, and with albendazole alone (all P values <0.05). The combination group showed better antilymphangiogenesis effect than the group of double dose bevacizumab. The albendazole-treated group showed reduced expression of VEGF-A, VEGF-C, TNF-alpha, and VEGFR-2 compared with corneas from the PBS group (P value <0.05 in all respective comparisons). Conclusion: Albendazole significantly decreased NV and LY in the cornea. This beneficial effect is additively enhanced when combined with bevacizumab treatment.

Correction: Prognosis for ocular toxocariasis according to granuloma location.

[This corrects the article DOI: 10.1371/journal.pone.0202904.].

Prognosis for ocular toxocariasis according to granuloma location.

PURPOSE: To determine the prognosis for ocular toxocariasis (OT) according to the location of the granuloma and to identify factors associated with its recurrence within 1 year. METHODS: OT patients were classified according to the granuloma lesion. After grouping the patients as posterior or peripheral, we compared sex, age, intraocular pressure, best corrected visual acuity (BCVA), degree of inflammation, immunoglobulin E, eosinophil profiles, recurrence, and complications in each group. We also identified factors associated with recurrence within 1 year. RESULTS: A total of 29 (61.70%) patients had granuloma at the periphery, and 18 (38.30%) patients had granuloma around the posterior pole. There were no significant differences in ocular or systemic evaluations except the initial BCVA. The mean decimal BCVA of the posterior pole granuloma group was worse than that of the peripheral granuloma group (p = 0.042). After treatment, the mean BCVA of the posterior pole granuloma group improved significantly (p = 0.019), and the final mean BCVA was not significantly different between the groups (p = 0.673). Multiple logistic regression analyses revealed that recurrence within a year was associated with age at diagnosis (p = 0.007). CONCLUSIONS: The initial BCVA of OT patients differed according to the location of the granuloma, but the BCVA after treatment was not significantly different between the groups. Younger age was associated with recurrence within 1 year.

The Effect of 0.5% Timolol Maleate on Corneal(Lymph)Angiogenesis in a Murine Suture Model.

PURPOSE: To investigate the anti(lymph)angiogenic and anti-inflammatory effects of 0.5% timolol maleate in a murine corneal suture model. METHODS: Corneal neovascularization and lymphangiogenesis were compared in groups of mice that underwent corneal suture and were subsequently administered a subconjunctival injection of 0.5% timolol maleate, dexamethasone, or phosphate-buffered saline (PBS). 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 and interleukin (IL)-6], vascular endothelial growth factor (VEGF)-A, VEGF-C, vascular endothelial growth factor receptor (VEGFR)-2, and VEGFR-3. RESULTS: When corneas from the timolol-treated group were compared to the PBS-treated group, we observed decreases in angiogenesis, lymphangiogenesis, and inflammatory infiltration in the timolol-treated group (P value <0.05 in all respective comparisons). Corneas from the timolol-treated group showed reduced expression of VEGF-A, VEGF-C, TNF-alpha, IL-6, VEGFR-2, and VEGFR-3 compared to corneas from the PBS group (P value <0.05 in all respective comparisons). CONCLUSION: Blocking adrenergic signaling in the cornea with 0.5% timolol maleate decreased corneal neovascularization and lymphangiogenesis.

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.

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 with PBS. The sorafenib group showed reduced expression of VEGFR-3 compared with dexamethasone (P=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.

Preoperative Mannitolization Can Decrease Corneal Endothelial Cell Damage After Cataract Surgery.

PURPOSE: To evaluate whether preoperative mannitolization can change ocular biometry and affect postoperative corneal endothelial cell density. METHODS: Bilateral sequential cataract surgery was performed in 38 patients. Preoperative mannitolization was done in one eye of each subject. We checked the change in preoperative ocular biometry before and after intravenous mannitolization. We compared the postoperative corneal endothelial cell density between eyes with mannitolization and without mannitolization at postoperative week 1, 2, 5, and 8. We evaluated the relationship between change in ocular biometry and change in postoperative corneal endothelial cells in eyes that underwent preoperative mannitolization. RESULTS: After mannitolization, eyes exhibited decreased intraocular pressure, axial length (AL), and vitreous chamber depth (VCD) and increased anterior chamber depth (ACD) and lens position (LP) compared to before mannitolization (p < 0.05). Preoperative mannitolization has a tendency to decrease the intraoperative use of phaco energy in eyes with moderate nucleosclerosis. Eyes with preoperative mannitolization showed less loss of postoperative corneal endothelial cells than eyes without preoperative mannitolization (p < 0.05). The ACD, LP, and AL changes by mannitolization were all negatively correlated with corneal endothelial cell loss (p < 0.05). CONCLUSION: Preoperative mannitolization can decrease postoperative loss of corneal endothelial cells. The protective effect of preoperative mannitolization on the corneal endothelium may be due to the decreased need for phaco energy and changes in ocular biometry such as ACD, AL, and LP.

The Association Between Aqueous Connective Tissue Growth Factor and the Severity of Age-related Cataracts as Graded by the Lens Opacities Classification System III.

PURPOSE: To evaluate the relationship between aqueous humor concentrations of connective tissue growth factor (CTGF) and the severity of age-related cataracts. MATERIALS AND METHODS: We conducted a prospective clinical study on 43 eyes of 43 patients with senile cataracts scheduled to undergo routine phacoemulsification surgery. Before surgery, all patients were graded for cataract severity using the Lens Opacities Classification System III in terms of four features: nuclear opalescence (NO), nuclear color (NC), cortical cataracts (C), and posterior sub-capsular cataracts (P). During surgery, aqueous humor samples were obtained from all patients, and sandwich enzyme-linked immunosorbent assays (ELISAs) were used to determine CTGF concentrations. To assess any relationship between cataract severity and CTGF levels of the aqueous humor, various correlation analyses and multiple linear regression were used. RESULTS: We found a positive correlation between the overall cataract grade and aqueous CTGF level (p < 0.05). In addition, four features of the cataract grade (nuclear opalescence, nuclear color, cortical cataract and posterior sub-capsular cataract) were positively correlated with the aqueous CTGF concentration (p < 0.05). The final regression model identified overall cataract grade as an independent predictor of increased CTGF levels in the aqueous humor (p < 0.05). CONCLUSIONS: CTGF tends to increase in the aqueous humor as the severity of age-related cataracts increases. Therefore, this cytokine may play an important role in the pathogenesis of age-related cataracts. Additional studies are required for clarification of this finding.

Stop-Chop versus Retro-Chop Nucleotomy During Phacoemulsification.

PURPOSE: To compare the outcomes of two nuclear fracture techniques used in phacoemulsification (stop-chop vs. retro-chop). METHODS: A total of 131 eyes from 111 patients with severe cataracts and nucleosclerosis (of grades 4-5) were included. The patients were randomly assigned to undergo stop-chop or retro-chop nucleotomy during phacoemulsification. The Drysdale manipulator was used as the chopping instrument in both groups. The intraocular phacoemulsification parameters including phacoemulsification power and phacoemulsification time were compared across the two groups. The following parameters were compared postoperatively at 1, 2, 5, and 8 weeks using unpaired t test: corrected distance visual acuity (CDVA), corneal endothelial cell density (CD), central corneal thickness (CT). RESULTS: There were significant differences in the phacoemulsification parameters between the groups. Retro-chop group showed less phacoemulsification time (p < 0.05) and less phacoemulsification power (p < 0.05) than stop-chop group. There was also a significant difference in the rate of corneal endothelial cell loss between the two groups (p < 0.05). In contrast, there were no significant differences in the postoperative change of CT or CDVA between the groups. CONCLUSION: Retro-chop nucleotomy with a Drysdale nucleus manipulator is an effective and safe method. It reduces both the use of intraoperative ultrasound energy, and early postoperative corneal endothelial cell loss compared to other nucleotomy techniques.