Involvement of Toll-like receptor 2 on human corneal epithelium during an infection of Pythium insidiosum.

BACKGROUND: Pythium insidiosum, a pathogenic oomycete, is a common causative organism of infectious corneal ulcer. Studying the innate immune response at the ocular surface is important for better understanding of the underlying pathogenesis and host defense against P. insidiosum infection. OBJECTIVE: The present study aims to investigate the role of Toll-like receptor (TLR)2 on human corneal epithelial cells (HCECs) in P. insidiosum infection. METHODS: Human embryonic kidney (HEK) cells were stimulated with either P. insidiosum zoospores or hyphae. NF-κB activation was determined by spectrophotometric measurement of secreted embryonic alkaline phosphatase (SEAP) levels. The role of TLR2 in P. insidiosum infection was studied in HCECs and monocyte derived macrophages (MDMs) using anti-TLR2 neutralizing antibody. The expression levels of pro-inflammatory cytokines were determined. RESULTS: Both P. insidiosum hypha and zoospore stimulated TLR2-dependent NF-κB activation in HEK-Blue™-hTLR2 cells in dose-dependent manner. IL-6 and IL-8, but not IL-1ß, were upregulated in HCECs after stimulation with P. insidiosum. Blockade of TLR2 on HCECs altered neither IL-6 nor IL-8 expressions. In contrast, the 3 cytokines were upregulated in the stimulated MDMs and the expression levels of IL-1ß and IL-8 but not IL-6 were attenuated in TLR2 blockade MDMs. CONCLUSIONS: P. insidiosum was recognized by human TLR2 on HEK cells. The mRNA expression levels of certain cytokines were dependent of TLR2 in P. insidiosum infected MDMs but not HCECs at early stage of infection.

Epithelial analysis of simple limbal epithelial transplantation in limbal stem cell deficiency by in vivo confocal microscopy and impression cytology.

Simple limbal epithelial transplantation (SLET) is a relatively new treatment for severe limbal stem cell deficiency. Outcomes of treatment are typically determined based on clinical manifestations. In this prospective-multicenter study, we aimed to analyze the epithelial phenotypes of the corneas after SLET using IVCM and IC, and correlated them with clinical findings. Ten eyes of nine patients, who underwent SLET (five autologous SLET and five living-related SLET) were recruited. A set of examinations included slit-lamp biomicroscopy, corneal in vivo confocal microscopy (IVCM), and impression cytology (IC) was performed in all eyes at least twice (≥ 3-month interval) postoperatively. Then, a correlation between findings of the three examinations was analyzed. There were seven eyes with clinical success (no central neovascularization) showed pure corneal epithelial phenotype or mixed corneal-conjunctival phenotypes (mostly cornea) in either IVCM or IC. Three eyes with clinical failure, presented with peripheral and central neovascularization, showed total or predominant conjunctival phenotype in IVCM and sole conjunctival phenotype in IC. From a total of 22 sets of examinations, there was a high correlation between clinical manifestation vs. IC (κ = 0.844, observed agreement = 81.82%) and a substantial correlation between clinical manifestation vs. IVCM (κ = 0.727, observed agreement = 76.19%) and between IVCM versus IC (κ = 0.729, observed agreement = 76.19%). In conclusion, IVCM and IC facilitate determination of epithelial phenotype of the cornea after SLET. There was a substantial to high correlation between IVCM, IC and clinical presentations. Findings observed by IVCM and IC may allow early detection of epithelial alterations in eyes underwent SLET before clinical recognition.

Long-term result of autologous cultivated oral mucosal epithelial transplantation for severe ocular surface disease.

The present study aimed to investigate the clinical outcomes of autologous cultivated oral mucosal epithelial transplantation (COMET) on human amniotic membrane (AM) for corneal limbal stem cell deficiency (LSCD). In this prospective, noncomparative case series, 20 eyes (18 patients) with bilateral severe ocular surface disease were chosen to undergo COMET on human AM. The primary outcome was clinical success, and the secondary outcomes were the best-corrected visual acuity difference, corneal opacification, symblepharon formation, and complications. The mean patient age was 48.2 ± 15.5 years. The mean follow-up time was 31.9 ± 12.1 months (range 8-50 months). All except one eye exhibited complete epithelialization within the first postoperative week. A successful clinical outcome, defined as a stable ocular surface without epithelial defects, a clear cornea without fibrovascular tissue invasion at the pupillary area, and no or mild ocular surface inflammation, was obtained in 15 of 20 eyes (75 %). The clinical success rate at 1 year was 79.3 %, and that at 4 years (end of follow-up) was 70.5 %. Fourteen of 20 (70 %) eyes exhibited improvement in visual acuity after COMET, and some required subsequent cataract surgery (2 eyes), penetrating keratoplasty (3 eyes), or keratoprosthesis implantation (1 eye). Preoperative symblepharon was eliminated in most eyes (8 of 13, 61.5 %) after COMET combined with eyelid reconstruction when needed. The only complication was corneal perforation (1 eye) induced by a severe eyelid abnormality; treatment with a tectonic corneal graft was successful. COMET can successfully restore ocular surface damage in most eyes with corneal LSCD.

Comparative analysis of long-term results of three epithelial cell transplantation procedures for treating limbal stem cell deficiency.

This study compared the long-term outcome of different epithelial transplantation techniques to treat limbal stem cell deficiency (LSCD). We conducted a retrospective 15-year comparative systematic cohort study of patients with LSCD who underwent either cultivated limbal epithelial transplantation (CLET), simple limbal epithelial transplantation (SLET), or cultivated oral mucosal epithelial transplantation (COMET). We reviewed the demographic data, etiology, LSCD severity, best-corrected visual acuity, surgical outcomes, and complications. A total of 103 eyes of 94 patients (mean age, 45.0 ± 16.4 years) with LSCD were enrolled. The most common cause of LSCD was chemical injury (42.7 %). The median follow-up time was 75 months. The success rates of CLET, SLET, and COMET were 45.5 %, 77.8 %, and 57.8 %, respectively. The 7-year survival rates after CLET, SLET, and COMET were 50.0 %, 72.2 %, and 53.2 %, respectively. Steven-Johnson syndrome (SJS) had a significantly lower survival rate than other causes (p < 0.001), but SLET had a significantly higher survival rate than CLET (p = 0.018) and COMET (p = 0.047). Visual improvement of more than four Snellen lines was achieved in 53.1 % of successful cases and 28.2 % of failed cases. SJS, Schirmer I test <5 mm, and the presence of postoperative recurrent epithelial defects were significant risk factors for a failed surgery. All epithelial transplantation techniques had favorable long-term surgical outcomes. More than half of the patients achieved a stable ocular surface and visual acuity improvement up to 7 years postoperatively. SLET tends to have a better surgical outcome than CLET and COMET, especially in patients with SJS.

Genetic analysis of allogenic donor cells after successful allo-limbal epithelial transplantation in simple and cultivated limbal epithelial transplantation procedures.

This non-comparative cohort study investigated long-term donor cell survival after allogenic simple/cultivated limbal epithelial transplantations (allo-SLET/allo-CLET, respectively) by genetic analysis. Transplanted corneal epithelial cells, which underwent impression cytology and/or corneal-button biopsy, were examined for personal identities of autosomal short-tandem repeats; the percentages of donor cells were calculated based on matching recipient or donor buccal-DNA references. Twelve patients were included; 4 underwent allo-CLET, 8 underwent allo-SLET. Eight patients (67%) had total limbal stem cell deficiency (LSCD). Genetic analysis was performed postoperatively (mean, 55.3 months). Donor cells were detected in 4 of 12 patients (25%), all of whom underwent allo-SLET; 1 patient had a donor genotype and 3 patients had a mixed donor/recipient genotype. The longest time of donor cell detection was 30 months. Seven patients (58%) used systemic immunosuppressives at the time of genetic analysis (mean use, 22.5 months). Allogenic donor cells survived in both procedures for the long term postoperatively, which encourages the long-term use of systemic immunosuppressives. Donor cells may not be the only factor in graft survival, in that most successful cases had a recipient profile. Their presence for a specific time may promote niches for the patients' own cells to repopulate, especially for partial LSCD.

Risk factors for contact lens related microbial keratitis: a case-control study.

OBJECTIVE: To evaluate the risk factors that may contribute to microbial keratitis in contact lens wearers. MATERIAL AND METHOD: This prospective case-control study included 52 patients with contact lens related keratitis treated between 1 December 2006 and 15 October 2007 at the Faculty of Medicine, Siriraj Hospital. Controls were sixty-three contact lens users attending with disorders unrelated to contact lens wear All subjects with informed consent were interviewed about demographic data, types of contact lens and contact lens solutions, contact lens use past the recommended replacement date, overnight wear, and lens hygiene. Odds ratio (OR, estimates of relative risks) and 95% confidence interval (CI) were calculated through multivariable logistic regression analysis. RESULTS: The use of contact lens past the replacement date caused the highest risk of developing microbial keratitis (OR = 9.1; CI 1.8-45.4, p = 0.005). Overnight wear of lenses (OR = 2.9, CI 1.3-6.2, p = 0.012) and poor lens hygiene (OR = 2.3, CI 1.0-5.1, p = 0.007) significantly increased the risk of microbial keratitis, respectively. None of the other risk factors showed a significant association. CONCLUSION: The significant risk factors for contact lens related microbial keratitis in the present study were the use of contact lens past the replacement date, overnight wear, and poor lens hygiene. Thus, the appropriate advice of contact lens care and usage may reduce the risk of microbial keratitis in contact lens wearers.

Chronic recalcitrant bacterial infection in steroid modified interstitial (stromal) keratitis: presentation and management.

OBJECTIVE: To report histopathologically proven bacterial infection manifested multifocal interstitial (stromal) keratitis (IK) with definite previous history of prolong topical steroid use. Standard managements of bacterial keratitis did not provoke enough benefit. MATERIAL AND METHOD: A retrospective analysis of 19 eyes in 15 patients referred to Siriraj Hospital between 2004 and 2010. RESULTS: Multifocal intrastromal infiltration, with relatively quiet ocular reaction and mild inflammation were initially presented in all eyes. They all previously had been diagnosed of presumed viral keratitis, and had been given topical corticosteroid treatment for a prolonged period of time without healing. Autoimmune disease workups were all negative. Corneal scrapings showed negative culture results in all eyes. However, bacteria within stromal lamellae with absent or minimal inflammatory cells were demonstrated in all eyes by corneal biopsies. In addition, cytology results obtained from 16S rDNA sequencing revealed Stenotrophomonas maltophilia in one eye and coagulase-negative staphylococci in two eyes. No case responded well to intensive topical and systemic antibiotics. However they were successfully treated with penetrating keratoplasty (11 eyes, 57.9%) or intrastromal antibiotic injections (8 eyes, 42.1%). CONCLUSION: Bacterial infection should be a concern in prolonged chronic IK. This was considered as primary bacterial IK or bacterial superinfection in immunocompromised cornea. Early recognition and appropriately aggressive managements contribute to successful outcome. Corneal biopsy is always essential and 16S rDNA sequencing is useful in this distinct clinical entity.

Refractive outcomes of femtosecond LASIK for myopic correction at Siriraj Hospital, Thailand.

OBJECTIVE: To evaluate refractive outcomes of myopic femtosecond LASIK at Siriraj Hospital, Thailand. MATERIAL AND METHOD: A retrospective study of patients who underwent femtosecond LASIK at SiLASIK center Siriraj hospital, Mahidol University, Thailand,from April 2009 to April 2010 was conducted. All patients had completely normal preoperative eye examination. All LASIK procedures were performed using Intralase femtosecond laser (Abbott Medical Optics Inc (AMO)) and VISX star S4 excimer laser (AMO). Postoperative follow-up included visual acuity (ETDRS chart) and manifest refraction at 1 day, 1 week, 1 month and 3 months. The flap thickness was evaluated at 3 months using Visante optical coherence tomography (OCT Carl Zeiss Meditec). RESULTS: One hundred and twenty eight eyes of 64 patients were included in the study. Mean preoperative manifest refraction spherical equivalent (MRSE) was -5.47 +/- 2.28 diopters (range, -1.88 to -12.63 diopters). At 3 months post-operation, the MRSE was -0.26 +/- 0.39 diopters (range, + 0.25 to -2.00 diopters) and 95.31% were within + 1.00 diopter of intended correction. There was 98% of eyes achieved uncorrected distance visual acuity (UDVA) 20/40 or better and 73% achieved UD VA 20/20 or better There was 31% of eyes gained one line and 15% lost 1 line of corrected distance visual acuity (CDVA). The mean error of flap thickness (difference between actual and intended central flap thickness) was + 0.23 +/- 1.77 microm for 120 microm flap setting and + 0.22 +/- 0.97 microm for 110 microm flap setting. The flap morphology was planar configuration. CONCLUSION: Myopic femtosecond LASIK at Siriraj Hospital provided satisfactory refractive outcomes and high accuracy of the flap thickness.

Comparison of epitheliotrophic factors in platelet-rich plasma versus autologous serum and their treatment efficacy in dry eye disease.

Current treatment of severe dry eye disease (DED) includes blood-derived eye drops, such as autologous serum (AS), which lubricate the eyes and provide factors that improve ocular surface and aid in wound healing. Recent studies indicated that platelet-rich plasma (PRP) was also effective. This study aims to compare the concentration and stability of epitheliotrophic factors in AS and PRP and their efficacy in DED patients. Epitheliotrophic factors of interest are epidermal growth factor (EGF), fibronectin, platelet-derived growth factor-AB (PDGF-AB), and transforming growth factor-beta1 (TGF-ß1). We determined that all epitheliotrophic factors were present in AS and PRP at baseline and did not decrease in concentrations in all storage conditions (4 °C for 1 week and at - 20 °C for 1 and 3 months). However, differences in concentrations in AS and PRP were observed. PRP was also shown not to be inferior to AS in terms of efficacy in DED treatment in a prospective randomized control trial which evaluated ocular surface disease index, dry eye questionnaire, ocular surface staining, tear breakup time, and Schirmer test at baseline and at 1-month follow-up. Therefore, with its shorter preparation time, PRP could be considered as an alternative to AS for the treatment of DED.

Association of HLA polymorphisms and acetaminophen-related Steven-Johnson syndrome with severe ocular complications in Thai population.

BACKGROUND/AIMS: To investigate the association of genetic polymorphisms of human leucocyte antigens (HLA) class I and II genes with acetaminophen-related Steven-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) who developed severe ocular complications (SOC) in the Thai population. METHODS: A prospective case-control study including 20 unrelated Thai acetaminophen-related SJS/TEN patients with SOC and 60 Thai healthy volunteers, recruited at three university hospitals in Bangkok, Thailand, from September 2014 to August 2019. HLA genes were analysed using PCR amplification followed by hybridisation with sequence-specific oligonucleotide (SSO) probes with bead-based typing kits. The carrier and gene frequencies of individual HLA alleles in patients were compared with those in control volunteers based on dominant assumption using Fisher's exact test. RESULTS: Among HLA class I polymorphisms, HLA-A*33:03, HLA-B*44:03 and HLA-C*07:01 were significantly associated with acetaminophen-related SJS/TEN and SOC with high ORs (95% CI, corrected p value; Pc) in carrier frequency of 5.4 (1.8 to 16.3, Pc=0.0274), 9.0 (95% CI 2.7 to 30.4, Pc=0.0034), and 9.3 (2.8 to 30.2, Pc=0.0022), respectively. There were no significant HLA class II associations with the disease after corrected for a total number of alleles tested. CONCLUSION: HLA-B*44:03 was strongly associated with acetaminophen-related SJS/TEN patients who developed SOC in Thai population. In addition, we also found moderate to strong associations with HLA-A*33:03 and HLA-C*07:01 suggesting their potential roles in the pathogenesis of SOC in acetaminophen-related SJS/TEN.

Assessing the Risk Factors For Diagnosed Symptomatic Dry Eye Using a Smartphone App: Cross-sectional Study.

BACKGROUND: Dry eye (DE) is a chronic inflammatory disease of the ocular surface of the eye that affects millions of people throughout the world. Smartphone use as an effective health care tool has grown exponentially. The "Dry eye or not?" app was created to evaluate the prevalence of symptomatic DE, screen for its occurrence, and provide feedback to users with symptomatic DE throughout Thailand. OBJECTIVE: The purpose of this study was to compare the prevalence of symptomatic dry eye (DE), blink rate, maximum blink interval (MBI), and best spectacle-corrected visual acuity (BSCVA) between people with and without symptomatic DE and to identify risk factors for symptomatic DE in Thailand. METHODS: This cross-sectional study sourced data from the "Dry eye or not?" smartphone app between November 2019 and July 2020. This app collected demographic data, Ocular Surface Disease Index (OSDI) score, blink rate, MBI, BSCVA, and visual display terminal (VDT) use data. The criterion for symptomatic DE was OSDI score ≥13. RESULTS: The prevalence of symptomatic DE among individuals using this smartphone app in Thailand was 85.8% (8131/9482), with the Northeastern region of Thailand having the highest prevalence, followed by the Northern region. Worse BSCVA (median 0.20, IQR 0.40; P=.02), increased blink rate (median 18, IQR 16; P<.001), reduced MBI (median 8.90, IQR 10.80; P<.001), female sex (adjusted OR 1.83; 95% CI 1.59-2.09; P<.001), more than 6 hours of VDT use (adjusted OR 1.59; 95% CI 1.15-2.19; P=.004), and lower than bachelor's degree (adjusted OR 1.30; 95% CI 1.03-1.64; P=.02) were significantly associated with symptomatic DE. An age over 50 years (adjusted OR 0.77; 95% CI 0.60-0.99) was significantly less associated with symptomatic DE (P=.04). CONCLUSIONS: This smartphone DE app showed that the prevalence of symptomatic DE in Thailand was 85.8%. Signs and risk factors could be also evaluated with this smartphone DE app. Screening for DE by this app may allow for the development of strategic plans for health care systems in Thailand.

Long-term Outcomes of Boston Type I Keratoprosthesis, and Efficacy of Amphotericin B and Povidone-Iodine in Infection Prophylaxis.

PURPOSE: To evaluate the long-term outcomes, anatomic retention, and complications of Boston type I keratoprosthesis (KPro). DESIGN: Retrospective, interventional case series METHODS: A retrospective chart review of implantations performed January 2008-December 2017 was conducted. Risk factors for anatomical retention and functional success were analyzed. The incidences of infections with and without antimicrobial medications were compared. RESULTS: Twenty-seven eyes of 26 patients were recruited. The most common indication for KPro surgery was failed penetrating keratoplasties (22 eyes, 81.5%). All patients had preoperative best-corrected visual acuity (BCVA) worse than 3/60. Over the mean follow-up of 83.4 ± 28.4 months, 15 eyes (55.6%) demonstrated improved BCVA. The anatomical retention rate was 88.9%, and the functional success rate was 44.4% (retained KPro with BCVA ≥ 3/60). Eyes with ocular surface disease (OSD) had significantly more complications than those without OSD. The most common complications were retroprosthetic membrane formation (15 eyes, 55.6%) and infection (13 eyes, 48.1%). Infectious keratitis was primarily caused by gram-positive bacteria, whereas endophthalmitis was chiefly caused by fungal infection. The infection incidence was significantly lower in eyes using topical 0.1% amphotericin B and 5% povidone iodine (P = .008 and .021, respectively). CONCLUSIONS: With its good retention rate and visual outcomes, Boston type I KPro could be an alternative treatment for patients with conventional penetrating keratoplasty failure, especially with appropriate patient selection and complication prevention. Standard prophylactic antibiotics with the addition of topical 0.1% amphotericin B and 5% povidone iodine might be optional effective regimens for infection prevention, especially in tropical countries.

Provocation of dry eye disease symptoms during COVID-19 lockdown.

To assess the impact of COVID-19 lockdown measures on dry-eye symptoms in a community-based population, a cross-sectional study was conducted during the first wave of the COVID-19 outbreak in Thailand. An online survey was distributed via social media between June and July 2020. The questionnaire elicited information on demographics, dry-eye symptoms, use of visual display terminals, and mental health status. There were 535 respondents. Thirty-seven percent reported having been diagnosed with dry-eye disease (DED). During the lockdown, the mean dry-eye symptom score (DESS) of overall participants dropped significantly from 81.6 ± 15.9 to 79.8 ± 17.4 (P < 0.001). The mean, daily, visual display terminal (VDT) usage increased from 10.55 ± 5.16 to 13.08 ± 5.65 h (P < 0.001). A negative correlation between age and VDT usage was observed in both the normal and lockdown situations. One-quarter of all participants had an abnormal mental health status. The female gender (OR 1.86; 95% CI 1.14-3.04) and increased VDT usage during the lockdown (OR 5.68; 95% CI 3.49-9.23) were independently associated with worsening dry-eye symptoms. The lockdown measures abruptly altered the behaviors and lifestyles of the overall population. Excessive exposure to VDTs were associated with deteriorated dry-eye symptoms, and it possibly contributed to the increased DED incidence in the surveyed population.

Successful Ocular Surface Reconstruction in Complete Ankyloblepharon With the Simple Oral Mucosal Epithelial Transplantation Technique: A Case Report.

PURPOSE: To report an outcome of a patient with complete ankyloblepharon successfully managed with simple oral mucosal epithelial transplantation (SOMET). METHODS: A 55-year-old woman presented with complete adhesion of both lids to the ocular surface as a complication from Stevens-Johnson syndrome. We performed 2-staged reconstructive surgeries: the first stage was to perform ankyloblepharon lysis and surface reconstruction with a mucosal graft on the palpebral area and an amniotic membrane on the bulbar area, and the second stage was to reconstruct the bulbar area with a transplantation of small pieces of oral mucosa (SOMET technique). Postoperatively, the patient was evaluated for ocular surface stability, recurrent symblepharon, in vivo confocal microscopy, and impression cytology with immunofluorescence staining. RESULTS: Complete epithelialization of cornea-like epithelium was observed within 6 weeks after SOMET was performed. The ocular surface was stable over 1 year. Both fornices remained deep. In vivo confocal microscopy showed cornea-like epithelium mixed with conjunctival epithelium, as confirmed with immunofluorescence staining, which revealed cytokeratin 3, cytokeratin 7, and cytokeratin 12 positivity. CONCLUSIONS: SOMET is a simple modified technique using minimal oral mucosal tissue to regenerate epithelialization for complicated ocular surface reconstruction such as a complete ankyloblepharon repair. Although there was evidence of conjunctival invasion, stable ocular surface and deep fornices can be achieved for further visual rehabilitative procedure.

Phenotypic Characterization of Corneal Epithelium in Long-Term Follow-Up of Patients Post-Autologous Cultivated Oral Mucosal Epithelial Transplantation.

PURPOSE: To analyze the phenotype of the corneal epithelium in patients with long-term follow-up who underwent autologous cultivated oral mucosal epithelial transplantation (COMET) using in vivo confocal microscopy (IVCM) and impression cytology with immunofluorescence staining (ICIF). METHODS: Thirteen eyes from patients with severe limbal stem cell deficiency, who underwent COMET at least 48 months before, were recruited in this noncomparative cohort study. After eye examination, IVCM and ICIF were performed. Clinical manifestations of the cornea were evaluated and compared with epithelial findings detected by IVCM and ICIF [cytokeratin (CK) 3, CK7, and CK12]. Two corneal buttons derived from patients receiving the corneal transplantation post-COMET were sent for immunohistochemistry (CK3, CK6, CK7, CK12, paired box gene 6, p63, zonula occludens-1, and integrin ß -1). RESULTS: The mean age of patients was 51.2 ± 20.6 years, and the mean follow-up time since COMET was 78.7 ± 16.3 months. Six of 13 eyes showed clinically successful COMET. In these eyes, IVCM demonstrated predominant cornea-like epithelium and ICIF reported positivity for CK3 and CK12, confirming the presence of oral mucosal and corneal epithelium. Meanwhile, 7 eyes showed total conjunctivalization, corresponding with substantial conjunctival epithelium detected by IVCM and positivity for conjunctival (CK7) and oral mucosal epithelial (CK3) markers detected by ICIF. The immunohistochemistry of corneal buttons stained positive for oral mucosal, corneal epithelial, and stem cell markers (CK3, CK12, and p63). CONCLUSIONS: In long-term follow-up of COMET, epithelium of successful patients demonstrated cornea-like phenotype, whereas failed cases revealed mainly conjunctival phenotype. However, there were evidences that oral mucosal epithelial cells remained across the cornea in both successful and failed COMET as detected by IVCM and ICIF.

Efficacy and outcome of simple limbal epithelial transplantation for limbal stem cell deficiency verified by epithelial phenotypes integrated with clinical evaluation.

PURPOSE: To evaluate the efficacy and outcome of simple limbal epithelial transplantation (SLET) for limbal stem cell deficiency (LSCD) using epithelial phenotype detection integrated with clinical manifestation. METHODS: This prospective multicenter study included patients with LSCD who underwent autologous SLET (autoSLET) and living-related allogenic SLET (Lr-alloSLET). All patients were assessed by slit-lamp biomicroscopy, in vivo confocal microscopy (IVCM), and impression cytology with immunofluorescence staining (ICIF) before and after surgery. The criteria for success were the presence of a clinically non-conjunctivalized cornea and corneal epithelium detected by IVCM or ICIF. Otherwise, the case would be considered a failure. Visual improvement and risk factors for SLET failure were analyzed. RESULTS: A total of 28 eyes of 26 patients (11 autoSLET and 17 Lr-alloSLET) were included. The median age was 53 years (range, 35-63), and the follow-up time was 29.5 months (range, 17.5-39.8). The overall survival rate was 89.3% at 2 years and 75.6% at 3 years with no difference between autoSLET and Lr-alloSLET (p = 0.24). Seven eyes subsequently underwent penetrating keratoplasty. Immunohistochemistry analysis showed that all corneal buttons had corneal epithelium and limbal stem cell markers. Visual improvement was achieved in both SLET groups (p < 0.001). Failed SLET developed between 5 and 32 months postoperatively. However, absolute risk factors for SLET failure were unidentified. CONCLUSION: The efficacy of autoSLET and Lr-alloSLET for LSCD was excellent. Limbal explants can regenerate and restore the corneal surface while maintaining the characteristics of limbal stem cells as shown by epithelial phenotype detection and immunohistochemistry integrated with clinical evaluation.

Eyeliner Induces Tear Film Instability and Meibomian Gland Dysfunction.

PURPOSE: To compare the degree of tear film instability and severity of meibomian gland dysfunction between subjects who use eyeliner and those who do not use eyeliner. METHODS: This cross-sectional study included 42 healthy volunteer women who had no dry eye symptoms (Ocular Surface Disease Index score < 13) and aged between 18 and 40 years. The subjects were classified into 2 groups: an eyeliner-use group (EL: regularly used eyeliner ≥3 d/wk and continuously used ≥6 mo) and a noneyeliner-use group as controls. A questionnaire for ocular surface symptoms using a visual analog scale was administered. Then, a number of eye tests were performed [grading of conjunctival inflammation, fluorescein tear breakup time, ocular surface fluorescein staining, Schirmer I, evaluation of meibomian gland (MG) function, detection of eyelid margin abnormalities, and Demodex detection]. RESULTS: Tear breakup time was significantly lower in the EL group compared with controls (3.0 ± 1.9 vs. 5.8 ± 2.1 s, P < 0.001). MG grading was significantly higher in the EL group than in controls (P = 0.004); higher grade (grades 2-3) was found in 85.7% of EL and 47.6% of controls. Meiboscore was also higher in EL than in controls (P = 0.001). Regarding the morphological changes in lid margin, only telangiectasia was detected significantly more in EL (28.6%) compared with controls (4.8%) (P = 0.041). Conjunctival inflammation was observed 4 times more in EL (66.7%) than in controls (14.3%), P = 0.001. Other outcomes included ocular surface symptoms and fluorescein staining scores, and Schirmer I and Demodex detection were not significantly different between both groups. CONCLUSIONS: The regular use of eyeliner induces tear film instability and MG dysfunction.

Characterization of limbal explant sites: Optimization of stem cell outgrowth in in vitro culture.

Simple limbal epithelial transplantation (SLET) and cultivated limbal epithelial transplantation (CLET) are proven techniques for treating limbal stem cell deficiency (LSCD). However, the precise regions that are most suitable for preparing explants for transplantation have not been identified conclusively. Accordingly, this in vitro study aimed at determining ideal sites to be selected for tissue harvest for limbal stem cell culture and transplantation. We evaluated cell outgrowth potential and the expression of stem cell markers in cultures from 48 limbal explants from five cadaveric donors. The limbal explants were generated from the three specific sites: Lcor (located innermost and adjacent to the cornea), Lm (middle limbus), and Lconj (located outermost adjacent to the conjunctiva). We found that explants from the Lconj and Lm sites exhibited higher growth potential than those from the Lcor site. Transcript encoding the stem cell marker and p63 isoform, ΔNp63, was detected in cells from Lm and Lconj explants; expression levels were slightly, though significantly (p-value < 0.05), higher in Lm than in Lconj, although expression of ΔNp63α protein was similar in cells from all explants. Differential expression of ATP-Binding Cassette Subfamily G Member 2 (ABCG2) did not reach statistical significance. Immunohistochemistry by indirect immunofluorescence analysis of limbus tissue revealed that the basal layer in explant tissue from Lconj and Lm contained markedly more stem cells than found in Lcor explant tissue; these findings correlate with a higher capacity for growth. Collectively, our findings suggest that explants from the Lconj and Lm sites should be selected for limbal cell expansion for both CLET and SLET procedures. These new insights may guide surgeons toward specific limbal sites that are most suitable for stem cell culture and transplantation and may ultimately improve treatment outcomes in the patients with LSCD.

Tear film change and ocular symptoms after reading printed book and electronic book: a crossover study.

PURPOSE: To evaluate and compare tear film changes and ocular symptoms after reading an electronic book (e-book) and a printed book. STUDY DESIGN: Clinical and experimental. METHODS: Crossover study was conducted in 30 healthy volunteers, some of whom read an e-book and others a printed book for 20 minutes and then switched the following week. Tear meniscus height (TMH), non-invasive break-up time (NIBUT), fluorescein break up time (FBUT), corneal and conjunctival staining score, and questionnaires about seven ocular symptoms were evaluated before and after reading by both reading methods. RESULTS: After reading an e-book, FBUT and NIBUT were significantly decreased (p<0.001for both). Similar to printed book readers (p=0.006, p=0.04, respectively). TMH and corneal and conjunctival staining score showed no significant differences in either group. Comparing the two groups, the e-book group showed more decrease in TMH, FBUT, and NIBUT (p>0.05). Ocular symptoms were significantly increased in both groups. The e-book group showed more increase in all symptoms, but only tearing (p=0.03) and burning sensation (p=0.02) were significantly different. CONCLUSIONS: Reading an e-book affected tear film instability and significantly increased burning sensation and tearing to a larger extend than reading a printed book.

Association of IKZF1 SNPs in cold medicine-related Stevens-Johnson syndrome in Thailand.

PURPOSE: Our meta-analysis of several ethnic groups (Japanese, Korean, Indian, Brazilian) revealed a significant genome-wide association between cold medicine-related SJS/TEN (CM-SJS/TEN) with severe ocular complications (SOC) and IKZF1 SNPs, suggesting that IKZF1 might be a potential marker for susceptibility to CM-SJS/TEN with SOC. In this study, we examined the association between CM-SJS/TEN with SOC and the IKZF1 SNPs in the Thai population. METHODS: 57 CM-SJS/TEN with SOC and 171 control samples were collected at Chulalongkorn University and Mahidol University. Genomic DNA samples were genotyped for the IKZF1 SNPs at Kyoto Prefectural University of Medicine in Japan using the TaqMan SNP genotyping assay. RESULTS: The four SNPs previously reported to be associated with CM-SJS/TEN with SOC in the Japanese were examined in the Thai samples. Although the number of Thai cases (n = 57) was small, a significant association between CM-SJS/TEN with SOC and IKZF1 SNPs which included rs4917014 (T vs G, OR = 2.9, p = 0.0012, Pc = 0.0049), rs4917129 (T vs C, OR = 2.8, p = 0.0026, Pc = 0.010) and rs10276619 (G vs A, OR = 1.8, p = 0.012, Pc = 0.048) was identified. CONCLUSION: In addition to the Japanese, Korean and Indian populations, Thai cases with CM-SJS/TEN and SOC were significantly associated with IKZF1 SNPs. With our previous report of the critical role of IKZF1 in mucocutaneous inflammation, these results suggest that IKZF1 is important in the pathogenesis of CM-SJS/TEN with SOC.