IgG4-related disease presenting as recurrent scleritis combined with optic neuropathy.

BACKGROUND: We report a case of atypical presentation of IgG4-related disease (IgG4-RD) with recurrent scleritis and optic nerve involvement. CASE PRESENTATION: A 61-year-old male presented with ocular pain and injection in his left eye for 2 months. Ocular examination together with ancillary testing led to the diagnosis of scleritis, which relapsed in spite of several courses of steroid treatment. After cessation of steroid, the patient complained of severe retro-orbital pain and blurred vision. His best corrected vision was count finger, the pupil was mid-dilated and a relative afferent pupillary defect was found. Funduscopic examination demonstrated disc swelling. Magnetic resonance imaging (MRI) showed enhancing soft tissue encasing the left globe, medial rectus muscle and optic nerve. Systemic work-up revealed multiple nodules in right lower lung and a biopsy showed histopathological characteristics of IgG4-RD. Long-term treatment with corticosteroids and a steroid-sparing agent (methotrexate) led to significant improvement in signs and symptoms with no recurrence for 2 years. CONCLUSIONS: This case highlights the significance of IgG4-RD in the differential diagnosis of recurrent scleritis. IgG4-RD may cause optic neuropathy resulting in visual loss. Early diagnosis and proper treatment can prevent irreversible organ damage and devastating visual morbidity.

Visual fatigue induced by watching virtual reality device and the effect of anisometropia.

The effect of small anisometropia on visual fatigue when using virtual reality (VR) devices was investigated. Participants (n = 34) visited three times. In the first visit, VR exposure (10 min) was conducted with the full correction of the refractive error of both eyes. Experimental anisometropia was induced by adding a + 1.0 dioptre spherical lens either on the dominant eyes in the second visit or on the non-dominant eyes in the third visit. At each visit, the participants played a predetermined video game using a head-mounted display VR for 10 min. Visual fatigue was assessed before and after playing VR game using the Virtual Reality Symptom Questionnaire (VRSQ) and high-frequency component of accommodative microfluctuation. Results showed that watching VR induced significant increase of VRSQ score, significant decrease in the maximum accommodation power and objective increase in visual fatigue. Experimental anisometropia induction either on the dominant or non-dominant eyes did not aggravate visual fatigue. Practitioner summary: Mild differences in refractive error (up to 1.0 dioptre) between both eyes do not significantly increase ocular fatigue by viewing virtual reality device (10 min). The impact of small anisometropia may be limited in developing a virtual reality device. Abbreviations: VR: virtual reality; VRSQ: virtual reality symptom questionnaire; HMD: head-mounted display; HFC: high-frequency component.

A case of severe flare reaction observed in HLA B27 associated acute anterior uveitis.

BACKGROUND: Anterior chamber flare reaction refers to the light reflection from the protein in aqueous humor. We report a case of very severe flare reaction observed in human leukocyte antigen (HLA)- B27 associated acute anterior uveitis (AAU). CASE PRESENTATION: An age 43 male patient visited the uveitis clinic complaining of decreased visual acuity in the right eye which developed 1 week before. The detailed ophthalmic examination revealed very severe flare reaction in the anterior chamber with diffuse conjunctival hyperemia in the right eye. Pupil margin and iris details were barely observable. Oral prednisolone 20 mg daily with topical 1% prednisolone acetate (Pred Forte, Allergan, CA) every 2 h and 1% topical cyclopentolate (Cyclogyl, Alcon, TX) three times daily were immediately prescribed. The next day, the flare reaction of the right eye decreased significantly and inflammatory cells in the anterior chamber were visible. Detailed fundus examination revealed no inflammatory signs on the retina and ciliary body. Later, the blood test revealed positive HLA B27 and autoantibodies against lupus anticoagulant with mild elevation of C reactive protein. There were no signs for ankylosing spondylitis. Continued treatment and tapering of topical 1% prednisolone acetate for 4 weeks led to the complete resolution of the anterior uveitis. CONCLUSIONS: We experienced HLA-B27 AAU with the feature of a very severe flare reaction. Conventional uveitis treatment was successful to acquire the complete resolution of the inflammation.

Comparison of cytotoxicities and anti-allergic effects of topical ocular dual-action anti-allergic agents.

BACKGROUND: To investigate the cytotoxicities of the topical ocular dual-action anti-allergic agents (alcaftadine 0.25%, bepotastine besilate 1.5%, and olopatadine HCL 0.1%) on human corneal epithelial cells (HCECs) and their anti-allergic effects on cultured conjunctival epithelial cells. METHODS: A Methylthiazolyltetrazolium(MTT)-based calorimetric assay was used to assess cytotoxicities using HCECs at concentrations of 10, 20 or 30% for exposure durations of 30 min, 1 h, 2 h, 12 h or 24 h. Cellular morphologies were evaluated by inverted phase-contrast and electron microscopy. Wound widths were measured 2 h, 18 h, or 24 h after confluent HCECs monolayers were scratched. Realtime PCR was used to quantify anti-allergic effects on cultured human conjunctival cells, in which allergic reactions were induced by treating them with Aspergillus antigen. RESULTS: Cell viabilities decreased in time- and concentration-dependent manners. Cells were detached from dishes and showed microvilli loss, cytoplasmic vacuoles, and nuclear condensation when exposed to antiallergic agents; alcaftadine was found to be least cytotoxic. Alcaftadine treated HCECs monolayers showed the best wound healing followed by bepotastine and olopatadine (p < 0.0001). All agents significantly reduced the gene expressions of allergic cytokines (IL-5, IL-25, eotaxin, thymus and activation-regulated chemokine, and thymic stromal lymphopoietin) and alcaftadine had the greatest effect (p < 0.0001 in all cases). CONCLUSIONS: Alcaftadine seems to have less side effects and better therapeutic effects than the other two anti-allergic agents tested. It may be more beneficial to use less toxic agents for patients with ocular surface risk factors or presumed symptoms of toxicity.

Toxic anterior segment syndrome-an updated review.

BACKGROUND: Toxic anterior segment syndrome (TASS) can be a rare complication of anterior segment surgery. Here we reviewed the most recent advances in the understanding of TASS. METHODS: English articles related to TASS were retrieved from "PubMed" using the following keywords; "toxic anterior segment syndrome" or "TASS". The authors of this paper reviewed all the retrieved literature and critical findings were summarized. RESULTS: The onset of TASS can vary from hours to months. The clinical manifestations are also variable. The causes of TASS are broad and continue to expand and could not be elucidated in over half of the reported cases. Prompt and thorough investigation to explore the causes of TASS is critical. Surgeons should be fully aware and updated regarding possible etiologies and make ceaseless efforts to prevent TASS. This effort begins with establishing TASS prevention protocols and regularly training surgical staff. Proper cleaning of surgical instruments is critical and should follow the guidelines set by The American Society of Cataract and Refractive Surgery TASS Task Force. When TASS occurs, sharing information with other ophthalmologists and reporting new causes is crucial for the prevention of outbreaks. CONCLUSIONS: Anterior segment surgeons should be reminded that TASS is mostly preventable by the establishment of TASS prevention protocols, regular surgical staff training and thorough adherence to recommendations for cleaning and sterilizing intraocular surgical instruments.

Clinical Significance of Biofilm on Silicone Tubes Removed From Patients With Nasolacrimal Duct Stenosis.

PURPOSE: To determine the relationship between the optical density of biofilms on silicone tubes and surgical outcomes. METHODS: A total of 43 silicone tubes from 33 patients with nasolacrimal duct stenosis were enrolled at 6 months after bicanalicular silicone tube intubation. The removed silicone tubes were divided into 2 segments. One segment of silicone tube was cultured while the other segment was used to measure optical density of biofilm. Each segment was divided into 3 pieces according to average normal human nasolacrimal anatomy. The first piece was located from puncta to lacrimal sac. The second piece was inside the nasolacrimal duct. The third piece was in the nasal cavity. Surgical outcome was evaluated at postoperative 12 months based on Munk score and fluorescein dye disappearance test. RESULTS: A total of 31 (72.1%) patients were surgically successful while 12 (27.9%) patients had surgical failure with persistent epiphora. In the second piece of silicone tube, the mean optical density of biofilm was 0.2654 nm in the surgical success group and 0.4472 nm in the surgical failure group. These results showed statistically significant (P < 0.01) difference. The most frequently isolated organism was Staphylococcus aureus in the surgical success group (7 of 31 patients, 22.6%). It was Pseudomonas aeruginosa in the surgical failure group (6 of 12 patients, 50%). CONCLUSION: This is the first study that determines the relationship between biofilm on silicone tube and surgical outcome. Biofilm formed on silicone tubes in nasolacrimal duct might cause surgical failure.

The Antibiofilm efficacy of nitric oxide on soft contact lenses.

BACKGROUND: To investigate the antibiofilm efficacy of nitric oxide (NO) on soft contact lenses. METHODS: Nitrite (NO precursor) release from various concentrations (0-1000 µM) of sodium nitrite (NaNO2, NO donor) was measured by Griess Assay. Cell viability assay was performed using human corneal epithelial cell under various concentration (0-1000 µM) of NaNO2. Biofilm formation on soft contact lenses was achieved by adding Staphylococcus aureus or Pseudomonas aeruginosa to the culture media. Various concentrations of NaNO2 (0-1000 µM) were added to the culture media, each containing soft contact lens. After incubation in NaNO2 containing culture media for 1, 3, or 7 days, each contact lens was transferred to a fresh, bacteria-free media without NaNO2. The bacteria in the biofilm were dispersed in the culture media for planktonic growth. After reculturing the lenses in the fresh media for 24 h, optical density (OD) of media was measured at 600 nm and colony forming unit (CFU) was counted by spreading media on tryptic soy agar plate for additional 18 h. RESULTS: Nitrite release from NaNO2 showed dose-dependent suppressive effect on biofilm formation. Most nitrite release from NaNO2 tended to occur within 30 min. The viability of human corneal epithelial cells was well maintained at tested NaNO2 concentrations. The bacterial CFU and OD showed dose-dependent decrease in the NaNO2 treated samples on days 1, 3 and 7 for both Staphylococcus aureus and Pseudomonas aeruginosa. CONCLUSIONS: NO successfully inhibited the biofilm formation by Staphylococcus aureus or Pseudomonas aeruginosa on soft contact lenses in dose-dependent manner.

Low recurrence rate of anchored conjunctival rotation flap technique in pterygium surgery.

BACKGROUND: To report the recurrence rate for an anchored conjunctival rotation flap technique in primary pterygium surgery. METHODS: Primary pterygium surgeries performed using anchored conjunctival rotation flap techniques (110 eyes in 110 patients) with a minimum follow-up of 12 months were reviewed. In this technique, a conjunctival flap is rotated to cover the bare sclera and suture-fixated with either 8-0 polyglactin (41 eyes) or 10-0 nylon (69 eyes). The recurrence rate was determined, and the two suture materials utilized were compared. RESULTS: The recurrence rate was 2.71% (3 cases in 110 eyes) when an anchored conjunctival rotation flap technique was used and patients were monitored for 26.40 ± 17.09 months. Interestingly, the recurrences were only observed in polyglactin-sutured eyes. No recurrence was detected in nylon-sutured eyes. No other complications were observed in either group. CONCLUSIONS: The anchored conjunctival rotation flap technique for pterygium surgery has a relatively low recurrence rate. Nylon suture-fixation of the flap was found to be superior to polyglactin suture-fixation in preventing recurrence.

Reversible opacification of hydrophobic acrylic intraocular lens- two cases report.

BACKGROUND: The opacification of the intraocular lens (IOL) can cause significant visual deterioration. It is known that opacity of hydrophobic acrylic IOLs is rare. We report 2 cases of reversible optic opacification of hydrophobic acrylic intraocular lenses (Tecnis ZCB00, Abbott), observed within 2 months after uneventful cataract surgery. CASE PRESENTATION: Case 1: Uneventful cataract surgery was performed on the left eye of an 86-year-old diabetic man with chronic open-angle glaucoma. A hydrophobic acrylic intraocular lens (IOL; Tecnis ZCB00, Abbott, Lake Bluff, IL) was implanted in the bag. Eye drops containing 0.5% levofloxacin and 1.0% prednisolone were used after surgery along with topical anti-glaucoma medications. At 7 weeks postoperative, cloudy, concentric IOL opacification developed, accompanied by decreased visual acuity and increased intraocular pressure. However, the opacification completely disappeared after 9 weeks. Case 2: Uneventful cataract surgery was performed on the left eye of a 72-year-old woman. A hydrophobic acrylic IOL (Tecnis ZCB00) was implanted in the bag. At 2 weeks postoperative, cloudy, concentric IOL opacification developed, accompanied by ocular discomfort. After 4 weeks, opacification and discomfort completely disappeared. CONCLUSIONS: We observed two cases of completely reversible opacification of hydrophobic acrylic IOLs. The exact nature of the transient opacity remains unclear, but an inflammatory origin cannot be completely ruled out.

The effect of Vaccinium uliginosum extract on tablet computer-induced asthenopia: randomized placebo-controlled study.

BACKGROUND: To investigate the alleviation effect of Vaccinium uliginosum extract (DA9301) on tablet computer-induced asthenopia. METHODS: This was a randomized, placebo-controlled, double-blind and parallel study (Trial registration number: 2013-95). A total 60 volunteers were randomized into DA9301 (n = 30) and control (n = 30) groups. The DA9301 group received DA9301 oral pill (1000 mg/day) for 4 weeks and the control group received placebo. Asthenopia was evaluated by administering a questionnaire containing 10 questions (responses were scored on a scales of 0-6; total score: 60) regarding ocular symptoms before (baseline) and 4 weeks after receiving pills (DA9301 or placebo). The participants completed the questionnaire before and after tablet computer (iPad Air, Apple Inc.) watching at each visit. The change in total asthenopia score (TAS) was calculated and compared between the groups RESULTS: TAS increased significantly after tablet computer watching at baseline in DA9301 group. (from 20.35 to 23.88; p = 0.031) However, after receiving DA9301 for 4 weeks, TAS remained stable after tablet computer watching. In the control group, TAS changes induced by tablet computer watching were not significant both at baseline and at 4 weeks after receiving placebo. Further analysis revealed the scores for "tired eyes" (p = 0.001), "sore/aching eyes" (p = 0.038), "irritated eyes" (p = 0.010), "watery eyes" (p = 0.005), "dry eyes" (p = 0.003), "eye strain" (p = 0.006), "blurred vision" (p = 0.034), and "visual discomfort" (p = 0.018) significantly improved in the DA9301 group. CONCLUSIONS: We found that oral intake of DA9301 (1000 mg/day for 4 weeks) was effective in alleviating asthenopia symptoms induced by tablet computer watching. TRIAL REGISTRATION: The study is registered at www.clinicaltrials.gov (registration number: NCT02641470, date of registration December 30, 2015).

Keratoplasty in the United States: A 10-Year Review from 2005 through 2014.

PURPOSE: To report evolving indications and preferred techniques of corneal transplantation in the United States. DESIGN: Retrospective review. METHODS: Annual reports from the Eye Bank Association of America on corneal graft distribution in the United States from 2005 through 2014 were reviewed. MAIN OUTCOME MEASURES: Number and percentage of corneal grafts distributed for various types of keratoplasty and their surgical indications in the United States. RESULTS: The total number of corneal transplants increased from 44 277 in 2005 to 46 513 in 2014. In the past decade, penetrating keratoplasty dramatically decreased (from 95% to 42%) and largely has been replaced by various lamellar keratoplasty (LK) techniques (from 5% to 58%). Descemet stripping (automated) endothelial keratoplasty was the most common (50%) type of corneal transplantation performed in the United Stated in 2014. The volume of Descemet membrane endothelial keratoplasty (DMEK) has been doubling every year since 2011 and accounted for 11% of total endothelial keratoplasties in 2014. There was a significant shift in indication for corneal transplantation, with Fuchs' endothelial dystrophy (22%) being the most common, followed by corneal edema occurring after cataract surgery (12%) in 2014. Eye banks supplied precut corneal grafts for 68% of LK techniques in 2014. CONCLUSIONS: In the United States, there has been a major shift in preferred keratoplasty techniques over the past decade, with a wide adoption of new LK techniques.

Femtosecond laser refractive surgery: small-incision lenticule extraction vs. femtosecond laser-assisted LASIK.

PURPOSE OF REVIEW: Small-incision lenticule extraction (SMILE) is a novel technique devised to correct refractive errors. SMILE circumvents excimer laser photoablation of cornea, as the stromal lenticule cut by femtosecond laser is removed manually. Smaller incisions and preservation of anterior corneal biomechanical strength have been suggested as some of the advantages of SMILE over femtosecond laser-assisted LASIK (FS-LASIK). In this review, we compared previous published results of SMILE and FS-LASIK. The advantage, efficacy and safety of SMILE are compared with FS-LASIK. RECENT FINDINGS: SMILE achieved similar efficacy, predictability and safety as FS-LASIK. Greater preservations of corneal biomechanical strength and corneal nerves were observed in SMILE when compared with LASIK or PRK. Additionally, the incidence of postoperative dry eye syndrome was found to be less problematic in SMILE than in FS-LASIK. SUMMARY: SMILE is a promising new surgery for refractive error correction. Prospective and retrospective studies of SMILE have shown that results of SMILE are similar to FS-LASIK. With advances in femtosecond laser technology, SMILE may gain greater acceptance in the future.

Retinal nerve fiber layer configuration in eyes with epiretinal membrane.

PURPOSE: To characterize configurations of the optic nerve head (ONH) and peripapillary retinal nerve fiber layer (RNFL) eyes with epiretinal membrane (ERM). METHODS: We reviewed medical records of consecutive patients with ERM who were examined between September 2012 and August 2013. Patients were categorized based on the severity of ERM on fundus imaging; patients with ERM but without retinal distortion were in group 1, and patients with ERM and retinal distortion were in group 2. Optic nerve head and RNFL parameters measured by spectral domain optical coherence tomography were assessed and compared between eyes with ERM and normal fellow eyes. RESULTS: There were no significant differences in ONH and RNFL parameters between ERM eyes and normal fellow eyes in group 1 (n = 12). In group 2 (n = 33), cup volume was smaller (p = 0.020), temporal quadrant RNFL thickness was greater by 18.8 ± 16.6 µm (p < 0.001), and RNFL peak angle was smaller by 18.7 ± 25.3 degrees in ERM eyes compared with fellow eyes (p = 0.001). In the 15 patients in group 2 who underwent ERM removal, temporal quadrant RNFL thickness decreased and the RNFL peak angle increased after the surgery (p = 0.008 and p = 0.001, respectively). CONCLUSIONS: The configurations of the ONH and peripapillary RNFL in eyes with ERM were different from those in fellow eyes. The characteristic configurations of RNFL such as small peak angle and temporal quadrant thickening may be considered when evaluating parameters as measured by spectral domain optical coherence tomography.

Late spontaneous posterior capsule rupture with single-piece hydrophobic acrylic intraocular lens dislocation.

In this study, we described and discussed the late onset spontaneous posterior capsule rupture with intraocular lens (IOL) dislocation years after uncomplicated cataract surgery and implantation of hydrophobic acrylic IOLs. Eight patients presented with spontaneous posterior capsule rupture and IOL dislocation 5-20 years after uncomplicated phacoemulsification and IOL (AcrySof, Alcon, US) implantation. None of the patients had undergone posterior capsulotomy in the past. Four of the patients admitted habitual eye rubbing. An intact and well-centered continuous curvilinear capsulotomy edge was observed in all cases. IOLs were dislocated or displaced behind the anterior capsulotomy with a significant decrease in vision. A large rupture with a curled edge of the broken posterior capsule was visible. Dislocated IOLs were removed, and a three-piece IOL was inserted in the sulcus in six cases and suture fixated to the sclera in two cases. Improved vision was achieved in all cases. Although the mechanism underlying this late complication is unclear, habitual eye rubbing or IOL design may play a role. Further investigation is needed to prevent this complication in the future.

Alginate Hydrogel Integrated with a Human Fibroblast-Derived Extracellular Matrix Supports Corneal Endothelial Cell Functionality and Suppresses Endothelial-Mesenchymal Transition.

Human corneal transplantation is still the only option to restore the function of corneal endothelial cells (CECs). Therefore, there is an urgent need for hCEC delivery systems to replace the human donor cornea. Here, we propose an alginate hydrogel (AH)-based delivery system, where a human fibroblast-derived, decellularized extracellular matrix (ECM) was physically integrated with AH. This AH securely combined with the ECM (ECM-AH) was approximately 50 µm thick, transparent, and permeable. The surface roughness and surface potential provided ECM-AH with a favorable microenvironment for CEC adhesion and growth in vitro. More importantly, ECM-AH could support the structural (ZO-1) and functional (Na+/K+-ATPase) markers of hCECs, as assessed via western blotting and quantitative polymerase chain reaction, which were comparable with those of a ferritic nitrocarburizing (FNC)-coated substrate (a positive control). The cell density per unit area was also significantly better with ECM-AH than the FNC substrate at day 7. A simulation test of cell engraftment in vitro showed that hCECs were successfully transferred into the decellularized porcine corneal tissue, where they were mostly alive. Furthermore, we found out that the endothelial-mesenchymal transition (EnMT)-inductive factors (Smad2 and vimentin) were largely declined with the hCECs grown on ECM-AH, whereas the EnMT inhibitory factor (Smad7) was significantly elevated. The difference was statistically significant compared to that of the FNC substrate. Moreover, we also observed that TGF-ß1-treated hCECs showed faster recovery of cell phenotype on the ECM. Taken together, our study demonstrates that ECM-AH is a very promising material for hCEC culture and delivery, which endows an excellent microenvironment for cell function and phenotype maintenance.

Nitric oxide: an old drug but with new horizons in ophthalmology-a narrative review.

Background and Objective: Based on basic knowledge and prior research on nitric oxide (NO), the potential of NO for treating eye diseases is reviewed, and the possibility of NO-based eye drops in clinical practice and the future potential of NO in ophthalmology are discussed. Methods: A PubMed search was performed for English-language original reports and reviews using the following key words: nitric oxide, eye, ocular, and drug. Key Content and Findings: NO is synthesized in the human body by NO synthase (NOS) from L-arginine or through enzyme-dependent reduction of dietary nitrate. Three types of NOS (eNOS, nNOS, and iNOS) are abundantly expressed in the eye under normal physiologic or pathologic conditions. The biological effect of NO in the eye is dose dependent. Low intraocular NO concentrations, produced by eNOS or nNOS, have various cellular effects, including vasodilation, intraocular pressure (IOP) regulation, and neuroprotection. iNOS induced under pathologic ocular conditions produces high NO concentrations in the local environment and mediates tissue inflammation, ocular cell apoptosis, and neurodegeneration. In particular, increased iNOS has been reported in glaucoma and retinal ischemic or degenerative diseases. NO plays a vital role in ocular injury. NO can facilitate ocular surface wound healing while eradicating pathogens such as bacteria and Acanthamoeba in chemical burns or infectious keratitis. Furthermore, NO has antifibrotic activity via the cyclic guanosine monophosphate (cGMP) signaling pathway. NO causes smooth muscle relaxation, which can be used to inhibit myopia progression in children. NO can be a stem cell modulator and may help in treating ocular stem cell disorders. Conclusions: Because of its diverse biologic effects, NO can be a key player in regulating ocular inflammation in various ocular diseases, aiding ocular surface wound healing, controlling IOP in glaucoma, alleviating retinal disease, and suppressing myopia progression. Although there remain limitations to the effective use of highly unstable state, gaseous NO, the role of NO in the field of ophthalmology can be greatly expanded through the development of novel NO donors and effective delivery platforms.

Effects of In Vitro Combination of Nitric Oxide Donors and Hypochlorite on Acanthamoeba castellanii Viability.

Purpose: To investigate the combined anti-Acanthamoeba effects of nitric oxide (NO) donors and hypochlorite to maximize amoebicidal outcomes while minimizing damage to human corneal epithelial cells (HCECs). Methods: Acanthamoeba castellanii and primary cultured HCECs and keratocytes were treated with sodium hypochlorite (NaOCl), NO donors (sodium nitroprusside [SNP] and sodium nitrite [NaNO2]), or a combination of hypochlorite and NO donors. The viability of A. castellanii, HCECs, and keratocytes was assessed. Minimal inhibitory concentration (MIC) and fractional inhibitory concentration of NaOCl and NO donors were determined. The activation of mammalian targets of rapamycin (mTOR) and ERK and the expression of nitrite reductase and Nrf2 were assessed in HCECs using Western blot analysis. The cysticidal effects of combined NaOCl and NO donors were also evaluated. Results: A dose-dependent toxicity was observed in A. castellanii, HCECs, and keratocytes when treated with NaOCl and SNP. The range of tested NaNO2 concentrations showed no significant toxicity to HCECs; however, dose-dependent toxicity to A. castellanii was observed. The MIC of NaOCl against HCECs and A. castellanii was 8.0 mg/mL. The MIC of NaNO2 and SNP was 500 mM and 10 mM in both HCECs and A. castellanii, respectively. Weak attenuation of the mTOR and ERK phosphorylation was observed and Nrf2 expression decreased slightly after exposure of HCECs to 2.0 mg/mL NaOCl. For the combination treatment, NaOCl (0.125 mg/mL) was selected based on the safety of HCECs and the toxicity of A. castellanii. A more potent anti-Acanthamoeba effect and HCEC toxicity were observed when NaOCl was combined with SNP rather than NaNO2. Conclusions: Combined NaOCl and NO donors had a stronger anti-Acanthamoeba effect compared to either drug alone. Translational Relevance: This study demonstrates that the combined use of various drugs for the treatment of Acanthamoeba infection can enhance the anti-Acanthamoeba effect while minimizing the toxicity of the individual drug.

Effect of Magnetic Microparticles on Cultivated Human Corneal Endothelial Cells.

Purpose: To investigate effects of magnetic microparticles on movement of magnet controlled human corneal endothelial cells (HCECs). Methods: Immortalized HCEC line (B4G12) and primary culture of HCECs were exposed to two commercially available magnetic micro- or nanoparticles, SiMAG (average size 100 nm) and fluidMAG (average size <1000 nm). Cell viability assays and reactive oxygen species production assays were performed. Cellular structural changes, intracellular distribution of microparticles, and expression levels of proteins related to cellular survival were analyzed. Ex vivo human corneas were exposed to microparticles to further evaluate their effects. Magnetic particle-laden HCECs were cultured under the influence of a neodymium magnet. Results: No significant decrease of viability was found in HCECs after exposure to both magnetic particles at concentrations up to 20 µg/mL for 48 hours. However, high concentrations (40 µg/mL and 80 µg/mL) of SiMAG and FluidMAG significantly decreased viability in immortalized HCECs, and only 80 µg/mL of SiMAG and FluidMAG decreased viability in primary HCECs after 48 hours of exposure. There was relative stability of viability at various concentrations of magnetic particles, despite a dose-dependent increase of reactive oxygen species, lactate dehydrogenase, and markers of apoptosis. Ex vivo human cornea study further revealed that exposure to 20 µg/mL of SiMAG and fluidMAG for 72 hours was tolerable. Endocytosed magnetic particles were mainly localized in the cytoplasm. The application of a magnetic field during cell culture successfully demonstrated that magnetic particle-loaded HCECs moved toward the magnet area and that the population density of HCECs was significantly increased. Conclusions: We verified short-term effects of SiMAG and fluidMAG on HCECs and their ability to control movement of HCECs by an external magnetic field. Translational Relevance: A technology of applying magnetic particles to a human corneal endothelial cell culture and controlling the movement of cells to a desired area using a magnetic field could be used to increase cell density during cell culture or improve the localization of corneal endothelial cells injected into the anterior chamber to the back of the cornea.

Viral transgene expression delivered by repeat intraocular adenoviral vector injection: in vivo live imaging study.

We delivered adenovirus vector (Ad) via intravitreous injection and monitored transgene (luciferase) expression in living mice (BALB/c) at multiple time points. In vivo live imaging technology was able to assess dynamically intraocular luciferase expression in a single animal population throughout the entire experiment period. Using this information, we were able to determine the optimal time point for readministration of Ad into the eyes and to dynamically study the time course of expression of a second Ad administration. Optical imaging demonstrated the limited period of transgene expression in eyes. Significant transgene signal was also detected in livers. The repeat intraocular delivery of the adenovirus resulted in significant blunting of transgene expression in both eyes and livers compared to the initial delivery. Periocular corticosteroid (triamcinolone acetonide) injection combined with initial Ad delivery was effective to rescue luciferase expression on repeat Ad vector delivery. However, this effect was not observed when corticosteroid was combined with repeat Ad delivery. Although corticosteroid enhanced ocular transgene expression, it also increased transgene expression in liver, which has potential safety implications. This dynamic transgene expression in eyes was successfully traced and monitored via a live imaging technique.