Mesenchymal stem/stromal cells precondition lung monocytes/macrophages to produce tolerance against allo- and autoimmunity in the eye.

Intravenously administered mesenchymal stem/stromal cells (MSCs) engraft only transiently in recipients, but confer long-term therapeutic benefits in patients with immune disorders. This suggests that MSCs induce immune tolerance by long-lasting effects on the recipient immune regulatory system. Here, we demonstrate that i.v. infusion of MSCs preconditioned lung monocytes/macrophages toward an immune regulatory phenotype in a TNF-α-stimulated gene/protein (TSG)-6-dependent manner. As a result, mice were protected against subsequent immune challenge in two models of allo- and autoimmune ocular inflammation: corneal allotransplantation and experimental autoimmune uveitis (EAU). The monocytes/macrophages primed by MSCs expressed high levels of MHC class II, B220, CD11b, and IL-10, and exhibited T-cell-suppressive activities independently of FoxP3(+) regulatory T cells. Adoptive transfer of MSC-induced B220(+)CD11b(+) monocytes/macrophages prevented corneal allograft rejection and EAU. Deletion of monocytes/macrophages abrogated the MSC-induced tolerance. However, MSCs with TSG-6 knockdown did not induce MHC II(+)B220(+)CD11b(+) cells, and failed to attenuate EAU. Therefore, the results demonstrate a mechanism of the MSC-mediated immune modulation through induction of innate immune tolerance that involves monocytes/macrophages.

Mooren Ulcer in a Child Wearing Orthokeratology Contact Lenses.

PURPOSE: To report a case of Mooren ulcer that developed in a pediatric patient wearing orthokeratology overnight contact lenses. METHODS: Case report. RESULTS: A 10-year-old boy was referred to our clinic because of progressive peripheral corneal ulcer in the right eye, despite the intensive use of fortified antibiotic eye drops. The patient had been using overnight orthokeratology lenses for 4 months before presentation of corneal ulcer. There was no other history of ocular or systemic trauma and disorders. Microbiological tests of the lesion were negative. Systemic evaluation showed no sign of rheumatologic disease. Under a diagnosis of Mooren ulcer, the patient was treated with topical and systemic corticosteroids. After four weeks of treatment, the patient's symptoms rapidly disappeared, and corneal ulcer was healed. The vision recovered to normal with the correction of with-the-rule astigmatism. CONCLUSIONS: Mooren ulcer can develop in pediatric patients wearing orthokeratology contact lenses. Given rapid progression of Mooren ulcer in a young population, early diagnosis and proper treatment are essential to prevent a devastating outcome.

Effect of Iris registration on visual outcome in wavefront-guided LASEK for myopic astigmatism.

PURPOSE: This study aimed to investigate the effect of iris registration (IR) on visual outcomes in wavefront-guided LASEK for myopic astigmatism. METHODS: The retrospective chart review was performed for wavefront-guided LASEK using VISX Star S4 in patients with myopic astigmatism (cylinder ≥ 1.00 diopter[D]). Eyes were divided into IR group (LASEK with IR at the time of surgery) and Non-IR group (LASEK without IR system + failed-IR engagement during LASEK). Visual acuity (VA), astigmatism, higher-order aberration (HOA), and contrast sensitivity were assessed preoperatively and 3 months postoperatively. The IR and Non-IR groups were subcategorized depending on the spherical equivalent (lower myopia ≤-5.00 D vs. higher myopia >-5.00 D) for the comparison of HOA changes. RESULTS: Postoperative uncorrected VAs showed no differences between IR (n = 30) and Non-IR (n = 46). In astigmatic vector analyses, no differences were noted in the mean magnitude of error and the mean angle of error between two groups. There were no differences in postoperative total HOA, spherical aberration (SA), coma, and trefoil between the groups, either. The total HOA and SA increased in both groups, while coma increased only in Non-IR. In higher myopia, ΔRMS of coma was smaller in IR. Preoperative and postoperative total HOA were linearly correlated in Non-IR, but not for IR. Contrast sensitivity of 12 cycles per degree improved in both groups. CONCLUSION: IR had similar outcomes to conventional trackers in wavefront-guided LASEK, with less tendency of inducing coma, especially in higher myopia.

Comparison of the anti-inflammatory effects of induced pluripotent stem cell-derived and bone marrow-derived mesenchymal stromal cells in a murine model of corneal injury.

BACKGROUND AIMS: Mesenchymal stromal cells (MSCs) offer tremendous potential for therapeutic applications for inflammatory diseases. However, tissue-derived MSCs, such as bone marrow-derived MSCs (BM-MSCs), have considerable donor variations and limited expandability. It was recently demonstrated that MSCs derived from induced pluripotent stem cells (iPSC-MSCs) have less pro-tumor potential and greater expandability of homogenous cell population. In this study, we investigated the anti-inflammatory effects and mechanism of iPSC-MSCs in a murine model of chemical and mechanical injury to the cornea and compared the effects with those of BM-MSCs. METHODS: To create an injury, ethanol was applied to the corneal surface in mice, and the corneal epithelium was removed with a blade. Immediately after injury, mice received an intravenous injection of (i) iPSC-MSCs, (ii) BM-MSCs or (iii) vehicle. Clinical, histological and molecular assays were performed in the cornea to evaluate inflammation. RESULTS: We found that corneal opacity was significantly reduced by iPSC-MSCs or BM-MSCs. Histological examination revealed that the swelling and inflammatory infiltration in the cornea were markedly decreased in mice treated with iPSC-MSCs or BM-MSCs. Corneal levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 were lower in iPSC-MSC- and BM-MSC-treated mice, compared with vehicle-treated controls. In contrast, iPSC-MSCs with a knockdown of the TNF-α stimulating gene (TSG)-6 did not suppress the levels of inflammatory cytokines and failed to reduce corneal opacity. CONCLUSIONS: Together these data demonstrate that iPSC-MSCs exert therapeutic effects in the cornea by reducing inflammation in part through the expression of TSG-6, and the effects are similar to those seen with BM-MSCs.

Long-term safety from transmission of porcine endogenous retrovirus after pig-to-non-human primate corneal transplantation.

BACKGROUND: The risk of xenozoonosis mainly by porcine endogenous retrovirus (PERV) has been considered as one of the main hurdles in xenotransplantation and therefore should be elucidated prior to the clinical use of porcine corneal grafts. Accordingly, an investigation was performed to analyze the infectivity of PERVs from porcine keratocytes to human cells, and the long-term risk of transmission of PERVs was determined using pig-to-non-human primate (NHP) corneal transplantation models. METHODS: The infectivity of PERVs from the SNU miniature pig keratocytes was investigated by coculture with a human embryonic kidney cell line. Twenty-two rhesus macaques underwent xenocorneal transplantation as follows: (i) group 1 (n=4): anterior lamellar keratoplasty (LKP) with freshly preserved porcine corneas, (ii) group 2 (n=5): anterior LKP with decellularized porcine corneas followed by penetrating keratoplasty (PKP) with allografts, (iii) group 3 (n=3): PKP under steroid-based immunosuppression, (iv) group 4 (n=4): PKP under anti-CD154 antibody-based immunosuppression, (v) group 5 (n=4): deep anterior LKP with freshly preserved porcine corneas under anti-CD40 antibody-based immunosuppression, and (vi) group 6 (n=2): PKP under anti-CD40 antibody-based immunosuppression. Postoperative blood samples were serially collected, and tissue samples were obtained from thirteen different organs at the end of each experiment. The existence of PERV DNA and RNA was investigated using PCR and RT-PCR. RESULTS: Using two independent in vitro infectivity tests, neither PERV pol nor pig mitochondrial cytochrome oxidase II was detected after 41 and 92 days of coculture, respectively. After xenocorneal transplantation, a total of 257 serial peripheral blood mononuclear cell samples, 34 serial plasma samples, and 282 tissue samples were obtained from the NHP recipients up to 1176 days post-transplantation. No PERV transmission was evident in any samples. CONCLUSIONS: Within the limits of this study, there is no evidence to support any risk of PERV transmission from porcine corneal tissues to NHP recipients, despite the existence of PERV-expressing cells in porcine corneas.

Anti-CD40 antibody-mediated costimulation blockade promotes long-term survival of deep-lamellar porcine corneal grafts in non-human primates.

BACKGROUND: Corneal xenotransplantation is an effective solution for the shortage of human donor corneas, and the porcine cornea may be a suitable candidate for the donor cornea because of its optical similarity with humans. However, it is necessary to administer additional immunosuppressants to overcome antigenic differences. We aimed to investigate the feasibility of porcine corneas with anti-CD40 antibody-mediated costimulation blockade in a clinically applicable pig-to-non-human primate corneal xenotransplantation model. METHODS: Five Chinese rhesus macaques underwent deep-lamellar corneal transplantation using clinically acceptable sized (7.5 mm diameter) porcine corneal grafts. The anti-CD40 antibody was intravenously administered on a programmed schedule. Graft survival, central corneal thickness, and intraocular pressure were evaluated. Changes in effector and memory T and B cell subsets and anti-αGal and donor-specific antibodies were investigated in the blood, and the changes in complement levels in the aqueous humor and blood were evaluated. Memory cell profiles in the anti-CD40 antibody-treated group were compared with those from the anti-CD154 antibody-treated group or rejected controls presented in our previous report. The changes in anti-αGal, non-αGal, and donor-specific antibodies after 6 months were compared with baseline values. RESULTS: Anti-CD40 antibody-mediated costimulation blockade resulted in the successful survival of xenocorneal grafts (>389, >382, >236, >201, and >61 days), with 80% reaching 6 months of survival. Injection of anti-CD40 antibody considerably reduced the infiltration of inflammatory cells into the grafts and significantly blocked the complement response in the aqueous humor (P=.0159, Mann-Whitney U test). Systemic expansion of central or effector memory T cells was abrogated in the anti-CD40 antibody-treated primates compared with those in the rejected controls (P<.05, Mann-Whitney U test) or those in the anti-CD154 antibody-treated primates (P>.05, Mann-Whitney U test). The levels of anti-αGal, non-αGal, and donor-specific antibodies at 6 months were not significantly increased compared with baseline levels (P>.05, Wilcoxon signed rank test). CONCLUSIONS: An anti-CD40 antibody-mediated blockade appears to be effective immunosuppressive approach for porcine corneal deep-lamellar xenotransplantation in primates.

Mesenchymal stem/stromal cells protect against autoimmunity via CCL2-dependent recruitment of myeloid-derived suppressor cells.

Exogenously administered mesenchymal stem/stromal cells (MSCs) suppress autoimmunity despite transient engraftment. However, the mechanism is unclear. In this study, we report a novel mechanism by which MSCs modulate the immune system by recruiting myeloid-derived suppressor cells in a mouse model of experimental autoimmune uveitis (EAU). Intravenous infusion of MSCs blocked EAU development and reduced Th1 and Th17 responses. Time course analysis revealed an increase of MHC class II(lo)Ly6G(-)Ly6C(hi)CD11b(+) cells in draining lymph nodes by MSCs. These Ly6C(hi)CD11b(+) cells suppressed CD4(+) cell proliferation and Th1/Th17 differentiation and induced CD4(+) cell apoptosis. Adoptive transfer of Ly6C(hi)CD11b(+) cells ameliorated EAU, whereas depletion of Ly6C(hi)CD11b(+) cells abrogated the effects of MSCs. 1.8% of MSCs were present in draining lymph nodes 1 d after infusion, and MSCs with CCL2 knockdown did not increase MHC class II(lo)Ly6G(-)Ly6C(hi)CD11b(+) cells and failed to attenuate EAU. Therefore, our findings demonstrate that MSCs suppress autoimmunity by recruiting myeloid-derived suppressor cells into sites of inflammation in a CCL2-dependent manner.

Conjunctival granuloma with necrosis associated with exposed suture in upper double lid masquerading as ocular surface squamous neoplasia: a case report.

BACKGROUND: This study reports two cases of conjunctival granuloma with necrosis caused by an exposed suture in the upper palpebral conjunctiva masquerading as ocular surface squamous neoplasia. CASE PRESENTATION: Two patients presented with chronic conjunctival ulcerative and granulomatous lesions on the superior bulbar conjunctiva that repeatedly recurred after the mass was removed. The pathologic findings revealed the absence of malignant cells and presence of many lymphocytes, plasma cells, and histiocytes. There was no evidence of acid-fast bacilli or fungal organisms. When a past history of blepharoplasty was established, microscopic examination revealed occult exposed suture tips. After the sutures were removed, the granuloma with necrosis was resolved within a month. CONCLUSION: For all conjunctival lesions in the superior bulbar conjunctiva, a thorough examination of the ocular adnexae which includes eyelid eversion should be performed. There should be a suspicion of foreign body or exposed suture material especially when there is a non-healing ulcer.

Outdoor Air Pollution and Pterygium in Korea.

We investigated relationships between outdoor air pollution and pterygium in Korean adults. This study includes 23,276 adults in population-based cross-sectional data using the Korea National Health and Nutrition Examination Survey 2008-2011. Pterygium was assessed using slit lamp biomicroscopy. Air pollution data (humidity, particulate matter with aerodynamic diameter less than 10 µm [PM10], ozone [O3], nitrogen dioxide [NO2], and sulfur dioxide levels [SO2]) for 2 years preceding the ocular examinations were acquired. Associations of multiple air pollutants with pterygium or pterygium recurrence after surgery were examined using multivariate logistic models, after adjusting for several covariates. Distributed lag models were additionally used for estimating cumulative effects of air pollution on pterygium. None of air pollution factors was significantly associated with pterygium or pterygium recurrence (each P > 0.05). Distributed lag models also showed that air pollution factors were not associated with pterygium or pterygium recurrence in 0-to-2 year lags (each P > 0.05). However, primary pterygium showed a weak association with PM10 after adjusting for covariates (odds ratio [OR] 1.23; [per 5 µg/m³ PM10 increase]; P = 0.023). Aging, male sex, and greater sun exposure were associated with pterygium, while higher education level and myopia were negatively associated with pterygium (each P ≤ 0.001). Male sex and myopia were negatively associated with pterygium recurrence (each P < 0.05). In conclusion, exposure to higher PM10 levels was associated with primary pterygium, although this study observed no significant association between air pollution and overall pterygium or pterygium recurrence in Korean adults.

Biophysico-functional compatibility of Seoul National University (SNU) miniature pig cornea as xenocorneal graft for the use of human clinical trial.

BACKGROUND: Xenocorneal transplantation is one of the solutions for shortage of donor cornea, and remarkable advances have been made in pig-to-rhesus studies from the immunological perspective. Most successful preclinical trials have been carried out with corneas of the Seoul National University (SNU) miniature pig (SNU pig, genetically unmodified) as donor tissues; however, there has been no biophysico-functional evaluation of the SNU pig cornea as a substitute for human cornea. The purpose of this study was to investigate the biophysical and functional compatibility of SNU pig cornea for use in human clinical trials. METHODS: Ninety-three eyeballs obtained from 51 SNU pigs were used to evaluate the physical properties and changes in porcine corneal endothelial cells (PCECs) depending on preservation time and storage condition before surgery, proliferative and functional characteristics of PCECs, and the microbiologic safety of porcine cornea. Corneal diameters and curvatures, axial length, anterior chamber depth, and central corneal thickness were measured and compared with previously reported human data. Corneal endothelial cell density (ECD) was serially measured with a confocal microscope during 7 days of preservation in the same storage solution used for human corneas. Corneal endothelial cell proliferation and immunofluorescence staining of Na- and K-dependent ATPase in PCECs were evaluated after 7 days of preservation. The corneoscleral rims of SNU pigs were cultured for gram-positive bacteria, gram-negative bacteria, and fungi to evaluate their microbiological safety. RESULTS: Corneal diameter and thickness in SNU pigs was larger than human and corneal curvature was flatter; however, they were within surgically operable ranges. Mean ECD (day 0) and ECD loss after 7 days of preservation were 2625 ± 81 cells/mm(2) and 7.60 ± 1.53%, respectively, which is comparable to human ECD and ECD loss in the same conditions. The ECD of SNU pigs was inversely decreased with aging (R(2) = 0.4034, P = 0.001), and the estimated ages of pigs whose mean ECD would be more than 2500 and 2200 cells/mm(2) or more were 48 and 72 months or less, respectively. Mean doubling time of the endothelial cells was 52 to 96 h depending on the method used. The Na- and K-dependent ATPase pump in SNU pig cornea was well maintained for 7 days. No cultured microorganisms were found upon using the modified European Eye Bank Association protocol, which included additional antiseptic management during the enucleation procedure. CONCLUSIONS: In conclusion, SNU pig cornea is feasible for xenocorneal transplantation using the same preservation protocol as human with respect to biophysical and functional properties and can be stored for up to 7 days for transplantation in human clinical trials. An age limitation of donor pigs may be required for qualified corneal products to be used in human trials.

Mesenchymal stem/stromal cells protect the ocular surface by suppressing inflammation in an experimental dry eye.

Dry eye syndrome (DES) is one of the most common ocular diseases affecting nearly 10% of the US population. Most of the currently available treatments are palliative, and few therapeutic agents target biological pathway of DES. Although DES is a multifactorial disease, it is well-known that inflammation in the ocular surface plays an important role in the pathogenesis of DES. Mesenchymal stem/stromal cells (MSCs) have been shown to repair tissues by modulating excessive immune responses in various diseases. Therefore, we here investigated the therapeutic potential of MSCs in a murine model of an inflammation-mediated dry eye that was induced by an intraorbital injection of concanavalin A. We found that a periorbital administration of MSCs reduced the infiltration of CD4(+) T cells and the levels of inflammatory cytokines in the intraorbital gland and ocular surface. Also, MSCs significantly increased aqueous tear production and the number of conjunctival goblet cells. Subsequently, corneal epithelial integrity was well-preserved by MSCs. Together, the results demonstrate that MSCs protect the ocular surface by suppressing inflammation in DES, and suggest that MSCs may offer a therapy for a number of ocular surface diseases where inflammation plays a key role.

Prophylactic removal and microbiological evaluation of calcified plaques after pterygium surgery.

PURPOSE: The aim of this study was to investigate microbiological characteristics of prophylactically removed calcified plaques developed after pterygium excision, and to evaluate risk factors for the growth of microorganisms. METHODS: Only exposed calcified plaques developed at the same site of previous pterygium excision were prospectively removed in 15 eyes of 14 patients. Plaques were completely removed, divided into small pieces and evaluated for microbiological identification. Underlying scleral defects were reconstructed using a conjunctival autograft, amniotic membranes and scleral patch grafts according to the size and depth of the defects. Based on the results of microbiologic cultures, eyes were divided into two groups and risk factors for microbial growth were analyzed. RESULTS: At surgery, the mean age of the patients was 71.2 ± 5.8 years and 71.4 % were females. The mean time interval between pterygium excision and calcified plaque removal was 19.3 ± 13.8 years. Six of 15 (40 %) removed plaques showed bacterial growth, and Stenotrophomonas maltophilia was the most frequently isolated microorganism. The size of calcified plaques was the only risk factor for culture-positive results (p = 0.045). Underlying scleral defects were successfully repaired without any serious complication. CONCLUSIONS: Microorganisms can be isolated from calcified plaques developed at the site of previous pterygium excision, and the size of plaques is the only risk factor for culture-positive results. To remove potential source of infection, prophylactic removal of calcified plaques and scleral surface reconstruction should be considered, especially when the plaques are exposed and large.

Recurrent late-onset fibrotic capsular block syndrome after neodymium-yttrium-aluminum-garnet laser anterior capsulotomy: a case report.

BACKGROUND: Capsular block syndrome is an uncommon complication that occurs after cataract surgery. It is characterized by capsular distension, anterior intraocular lens displacement, anterior chamber shallowing, and unexpected myopic shifts. We report a case of recurrent fibrotic capsular block syndrome with Elschnig's pearl-type posterior capsule opacification 10 months after neodymium-yttrium-aluminum-garnet (Nd:YAG) laser anterior capsulotomy. CASE PRESENTATION: A 72-year-old Asian man complained of decreased visual acuity 5 years after undergoing phacoemulsification with posterior chamber lens implantation. Under slit-lamp examination, late postoperative capsular block syndrome was diagnosed and Nd:YAG laser anterior capsulotomy was performed. Ten months after anterior capsulotomy, the patient returned with decreased visual acuity and was diagnosed with recurrent fibrotic capsular block syndrome. Nd:YAG laser posterior capsulotomy was performed. CONCLUSIONS: We found that fibrotic capsular block syndrome could recur with Elschnig's pearl-type posterior capsule opacification after Nd:YAG laser anterior capsulotomy for late postoperative capsular block syndrome without posterior capsule opacification.

Corneal keloid: four case reports of clinicopathological features and surgical outcome.

BACKGROUND: Surgical outcome of corneal keloid is largely variable depending on reports, although surgical management is inevitable in visually significant cases. We here report clinical features, histopathological findings, and surgical outcome of four cases of corneal keloid. CASE PRESENTATION: Four Korean male patients without a history of corneal trauma or disease were clinically and histologically evaluated for a slowly-growing, white opacity in the cornea. On slit lamp examination, corneal lesions appeared as a solitary, pearly white, well-circumscribed nodule with a smooth and glistening surface. Because the lesions involved the visual axis deteriorating the visual acuity, the nodules were surgically removed by superficial keratectomy in all patients. Amniotic membrane transplantation was combined in three patients, and an intraoperative mitomycin C application in two patients. Hematoxylin-eosin staining of the excised nodules revealed epithelial hyperplasia, Bowman's layer disruption, thick and irregularly-arranged collagen fibers in the stroma, and accumulation of prominent fibroblasts, which are consistent with the diagnosis of corneal keloid. The corneal keloids recurred in all patients within 10 months of surgical excision and outgrew the boundary of the excised area. CONCLUSION: A diagnosis of corneal keloid should be suspected in patients presenting with an enlarging, white, glistening corneal nodule, even in the absence of a history of corneal trauma or disease. The recurrence is common after surgical excision, and the lesion can be exacerbated by surgery.

Estimation of axial curvature of anterior sclera: correlation between axial length and anterior scleral curvature as affected by angle kappa.

BACKGROUND: Though the development and fitting of scleral contact lenses are expanding steadily, there is no simple method to provide scleral metrics for scleral contact lens fitting yet. The aim of this study was to establish formulae for estimation of the axial radius of curvature (ARC) of the anterior sclera using ocular biometric parameters that can be easily obtained with conventional devices. METHODS: A semi-automated stitching method and a computational analysis tool for calculating ARC were developed by using the ImageJ and MATLAB software. The ARC of all the ocular surface points were analyzed from the composite horizontal cross-sectional images of the right eyes of 24 volunteers; these measurements were obtained using anterior segment optical coherence tomography for a previous study (AS-OCT; Visante). Ocular biometric parameters were obtained from the same volunteers with slit-scanning topography and partial coherence interferometry. Correlation analysis was performed between the ARC at 8 mm to the axis line (ARC[8]) and other ocular parameters (including age). With ARC obtained on several nasal and temporal points (7.0, 7.5, 8.0, 8.5, and 9.0 mm from the axis line), univariate and multivariate linear regression analyses were performed to develop a model for estimating ARC with the help of ocular biometric parameters. RESULTS: Axial length, spherical equivalent, and angle kappa showed correlations with temporal ARC[8] (tARC[8]; Pearson's r = 0.653, -0.579, and -0.341; P = 0.001, 0.015, and 0.015, respectively). White-to-white corneal diameter (WTW) and anterior chamber depth (ACD) showed correlation with nasal ARC[8] (nARC[8]; Pearson's r = -0.492 and -0.461; P = 0.015 and 0.023, respectively). The formulae for estimating scleral curvatures (tARC, nARC, and average ARC) were developed as a function of axial length, ACD, WTW, and distance from the axis line, with good determinant power (72 - 80 %; SPSS ver. 22.0). Angle kappa showed strong correlation with axial length (Pearson's r = -0.813, P <0.001), and the different correlation patterns of nasal and temporal ARC with axial length can be explained by the ocular surface deviation represented by angle kappa. CONCLUSIONS: Axial length, ACD, and WTW are useful parameters for estimating the ARC of the anterior sclera, which is important for the haptic design of scleral contact lenses. Angle kappa affects the discrepancies between the nasal and temporal scleral curvature.

Effect of Hydroxychloroquine Treatment on Dry Eyes in Subjects with Primary Sjögren's Syndrome: a Double-Blind Randomized Control Study.

The effect of hydroxychloroquine (HCQ) on dry eye has not been fully determined. This study aimed to compare the 12-week efficacy of HCQ medication with that of a placebo in the management of dry eye in primary Sjögren's syndrome (pSS). A double-blind, randomized control study was conducted in 39 pSS subjects from May 2011 through August 2013. pSS was diagnosed based on the classification criteria of the American-European Consensus Group. Subjects received 300 mg of HCQ or placebo once daily for 12 weeks and were evaluated at baseline, 6, and 12 weeks, with a re-visit at 16 weeks after drug discontinuance. The fluorescein staining score, Schirmer test score, tear film break-up time (TBUT), and ocular surface disease index (OSDI) were measured, and tears and blood were collected for ESR, IL-6, IL-17, B-cell activating factor (BAFF), and Th17 cell analysis. Color testing was performed and the fundus was examined to monitor HCQ complications. Twenty-six subjects completed the follow-up. The fluorescein staining score and Schirmer test score did not differ significantly. The OSDI improved with medication in the HCQ group but was not significantly different between the groups. TBUT, serum IL-6, ESR, serum and tear BAFF, and the proportion of Th17 cells did not change in either group. HCQ at 300 mg daily for 12 weeks has no apparent clinical benefit for dry eye and systemic inflammation in pSS (ClinicalTrials.gov. NCT01601028).

Mesenchymal stem/stromal cells inhibit the NLRP3 inflammasome by decreasing mitochondrial reactive oxygen species.

Mesenchymal stem/stromal cells (MSCs) control excessive inflammatory responses by modulating a variety of immune cells including monocytes/macrophages. However, the mechanisms by which MSCs regulate monocytes/macrophages are unclear. Inflammasomes in macrophages are activated upon cellular "danger" signals and initiate inflammatory responses through the maturation and secretion of proinflammatory cytokines such as interleukin 1ß (IL-1ß). Here we demonstrate that human MSCs (hMSCs) negatively regulate NLRP3 inflammasome activation in human or mouse macrophages stimulated with LPS and ATP. Caspase-1 activation and subsequent IL-1ß release were decreased in macrophages by direct or transwell coculture with hMSCs. Addition of hMSCs to macrophages either at a LPS priming or at a subsequent ATP step similarly inhibited the inflammasome activation. The hMSCs had no effect on NLRP3 and IL-1ß expression at mRNA levels during LPS priming. However, MSCs markedly suppressed the generation of mitochondrial reactive oxygen species (ROS) in macrophages. Further analysis showed that NLRP3-activated macrophages stimulated hMSCs to increase the expression and secretion of stanniocalcin (STC)-1, an antiapoptotic protein. Addition of recombinant protein STC-1 reproduced the effects of hMSCs in inhibiting NLRP3 inflammasome activation and ROS production in macrophages. Conversely, the effects of hMSCs on macrophages were largely abrogated by an small interfering RNA (siRNA) knockdown of STC-1. Together, our results reveal that hMSCs inhibit NLRP3 inflammasome activation in macrophages primarily by secreting STC-1 in response to activated macrophages and thus by decreasing mitochondrial ROS.

Adverse effects of low-dose systemic cyclosporine therapy in high-risk penetrating keratoplasty.

PURPOSE: The purpose of this study was to investigate the adverse effects of low-dose oral cyclosporine (CsA) therapy following high-risk corneal transplantation. METHODS: The medical records from 88 subjects who had undergone high-risk penetrating keratoplasties and had been administered oral CsA were retrospectively analyzed. High risk was defined as a history of graft rejection, three or more quadrants of vascularization, or the presence or history of intraocular inflammation. An initial CsA dose of 3-5 mg/kg per day was given for 3-7 days, followed by 2.5-3.5 mg/kg per day for approximately 1 month. The concentration of CsA was maintained at the target trough level of 120-150 ng/ml for at least 6 months or until serious complications developed. The relationship between the cumulative dose and duration of CsA administration and the adverse systemic effects, including the frequency of herpes keratitis, was evaluated. The incidence of herpes keratitis in the study subjects was compared with the incidence in 185 patients who had not received CsA therapy following penetrating keratoplasty. RESULTS: The mean survival time of the grafts was 33.6 months. Adverse effects occurred in 81.8 % of subjects. Hypertension, elevated liver enzyme levels, elevated serum creatinine level, and decreased absolute neutrophil count (ANC) were observed in 14.8, 6.8, 5.7, and 5.7 % of subjects, respectively. Simvastatin-induced rhabdomyolysis also developed in one case. Some patients exhibited minor complications, with gastrointestinal problems and hypertrichosis recorded in 5.7 and 3.4 % of subjects, respectively. Hypertension and hepatotoxicity most frequently occurred after 4 to 8 weeks of medication, while ANC decrease and nephrotoxicity generally developed after 24 weeks of treatment, with incidence related to the cumulative dose. Herpes keratitis occurred more frequently (31.8 %) in the CsA-treated subjects than in subjects that did not receive CsA therapy (p = 0.005). Most of the adverse effects were reversed after discontinuation of CsA therapy. CONCLUSION: The results of this study suggest that low-dose oral CsA therapy may induce various adverse effects, the most common of which are herpes keratitis and hypertension.

The pattern of early corneal endothelial cell recovery following cataract surgery: cellular migration or enlargement?

PURPOSE: To evaluate whether cellular migration or enlargement is the main mechanism of initial endothelial cell recovery following cataract surgery. METHODS: A prospective observational study, of 24 patients aged 50-80 years who were diagnosed with moderate cataract and received uncomplicated cataract surgery with a 2.75 mm temporal clear corneal incision, was performed in Seoul National University Bundang Hospital. Endothelial cell density (ECD) and area (ECA) were obtained in central and four paracentral (superior, inferior, nasal, and temporal) areas using non-contact specular microscopy. ECD, ECA, ECD% (ECD% = ECD in each area/the sum total of ECD in five areas), and the coefficient of variation of ECA (CV) in each location were investigated pre- and 1 day, 1 week, and 4 weeks postoperatively. RESULTS: ECD significantly decreased 1 day, 1 week, and 4 weeks postoperatively (p = 0.010, 0.015, and 0.003 respectively), and ECA increased (p = 0.008, 0.013, and 0.002 respectively) in only the temporal area. Postoperative ECD% decreased, and CV increased in only the temporal area significantly, when compared to preoperative values. There were no significant postoperative changes of ECD, ECA, ECD%, and CV in other areas. CONCLUSIONS: Postoperative changes of ECD, ECA, ECD%, and CV were limited to the temporal area adjacent to the primary corneal incision. Cellular enlargement, rather than migration, may have the major effect on early endothelial cell recovery after cataract surgery.

Conjunctival metaplasia after pterygium excision and limbal autograft.

PURPOSE: To investigate the changes of conjunctival epithelium in the pterygium and donor graft sites after pterygium excision and limbal conjunctival autograft. METHODS: This study included 16 eyes of 15 patients who underwent pterygium excision and limbal conjunctival autograft. Epithelial impression cytology specimens of both the pterygium and donor graft were obtained preoperatively and at months 1, 3, and 6, at the pterygium and donor graft sites, respectively. In each specimen, changes in the conjunctival epithelium, including the nucleus-to-cytoplasm (N/C) ratio and goblet cell density (GCD), were evaluated. Morphologic changes in the cells and nuclei were also evaluated. RESULTS: Preoperatively, both N/C ratio and GCD were significantly higher in specimens from the pterygium than in those from the donor site. At both sites, GCD decreased rapidly at 1 month after surgery and then gradually recovered. No significant difference in GCD was found between the two sites at 1, 3, and 6 months postoperatively. Although there was no significant difference in the N/C ratio at 1 and 3 months, the N/C ratio at the pterygium site was significantly greater than that at the donor site at 6 months. At 6 months postoperatively, changes suggesting squamous metaplasia, including elongation of the cells and pyknotic changes in the nuclei, were noted in five samples (31.3%) from the pterygium site but not in any of the samples obtained from the donor site. CONCLUSIONS: Conjunctival epithelial metaplasia may return after pterygium removal, which may be associated with the high rate of pterygium recurrence.