Interaction of Commonly Used Oral Molecular Excipients with P-glycoprotein.

P-glycoprotein (P-gp) plays a critical role in drug oral bioavailability, and modulation of this transporter can alter the safety and/or efficacy profile of substrate drugs. Individual oral molecular excipients that inhibit P-gp function have been considered a mechanism for improving drug absorption, but a systematic evaluation of the interaction of excipients with P-gp is critical for informed selection of optimal formulations of proprietary and generic drug products. A library of 123 oral molecular excipients was screened for their ability to inhibit P-gp in two orthogonal cell-based assays. ß-Cyclodextrin and light green SF yellowish were identified as modest inhibitors of P-gp with IC50 values of 168 µM (95% CI, 118-251 µM) and 204 µM (95% CI, 5.9-1745 µM), respectively. The lack of effect of most of the tested excipients on P-gp transport provides a wide selection of excipients for inclusion in oral formulations with minimal risk of influencing the oral bioavailability of P-gp substrates.

Are there Clinical Ways to Assess Inflammation in Dry Eye Disease?

In the diagnostic process of dry eye disease, the detection of inflammatory activity is critical in order to evaluate the risk of progression and immunologic shift of the disease, to predict patient response to treatment, and to design an efficient therapeutic strategy, including artificial tear replacement, punctal occlusion or anti-inflammatory therapy.Even if it is difficult to quantify, some indicators of the presence of inflammation are collectible during the examination of the ocular surface in a first-line clinical setting. This review presents and critically discusses the assessment of inflammation in dry eye disease in clinical practice.

New Freeware for Image Analysis of Lissamine Green Conjunctival Staining.

PURPOSE: Lissamine green (LG) is often used in addition to fluorescein to assess the severity of conjunctival damage in dry eye syndrome, which is graded manually. Our purpose was to describe an algorithm designed for image analysis of LG conjunctival staining. METHODS: Twenty pictures of patients suffering from dry eye with visible LG conjunctival staining were selected. The images were taken by 2 different digital slit lamps with a white light source and a red filter transmitting over the wavelengths absorbed by LG. Conjunctival staining appeared in black on a red background. The red channel was extracted from the original image. Stained areas were then detected using a Laplacian of Gaussian filter and applying a threshold whose value was determined manually on a subset of images. The same algorithm parameters remained constant thereafter. LG-stained areas were also drawn manually by 2 experts as a reference. RESULTS: The delineation obtained by the algorithm closely matched the actual contours of the punctate dots. In 19 cases of 20 (95%), the algorithm found the same Oxford grade as the experts, even for confluent staining that was detected as a multitude of dots by the algorithm but not by the experts, resulting in a high overestimation of the total number of dots (without mismatching the Oxford grade estimated by the experts). The results were similar for the 2 slit-lamp imaging systems. CONCLUSIONS: This efficient new image-analysis algorithm yields results consistent with subjective grading and may offer advantages of automation and scalability in clinical trials.

Antimicrobial activity of topical dyes used in clinical veterinary ophthalmology.

OBJECTIVE: To evaluate in vitro the antibacterial effects of fluorescein, rose bengal, and lissamine green topical ophthalmic dyes against selected Gram-positive and Gram-negative bacteria, and to evaluate whether preserved or preservative-free fluorescein solutions are able to inhibit or potentiate bacterial growth. PROCEDURES: Susceptibility testing was performed using the Kirby-Bauer disk diffusion method plated with clinical ocular isolates of Staphylococcus aureus, Staphylococcus pseudintermedius, Streptococcus spp., Escherichia coli, and Pseudomonas aeruginosa. Bacterial growth inhibition was evaluated 24 hours following the addition of commercially available fluorescein, rose bengal, and lissamine green sterile strips. Antimicrobial effectiveness testing was performed by inoculation of compounded 1% dye solutions, both with and without preservatives (fluorescein and lissamine contained thiomersal, and rose bengal contained nipagin and nepazol), with the five previously mentioned bacteria. Growth was evaluated at days 7, 14, and 28. RESULTS: All dyes showed antibacterial activity against Gram-positive organisms. Preservative-free compounded 1% fluorescein solution inhibited growth of Gram-positive organisms but not of Gram-negative organisms. Preservative-free rose bengal and lissamine green inhibited growth of both types of organisms. CONCLUSIONS: Preferably, ocular surface samples for antimicrobial culture should be taken prior to the administration of topical dyes, due to their potential antibacterial activity, particularly if undiluted strips are applied directly or commercial fluorescein solutions are used and not immediately rinsed. Ophthalmic dye solutions containing preservative are safe from bacterial growth for up to 28 days if properly handled and stored. The use of preservative-free fluorescein solutions should be avoided and preservative-free rose bengal and lissamine green should be handled carefully.

Comprehensive Clinical, Diagnostic, and Advanced Imaging Characterization of the Ocular Surface in Spontaneous Aqueous Deficient Dry Eye Disease in Dogs.

PURPOSE: To perform a comprehensive clinical, diagnostic, and imaging characterization of the ocular surface in West Highland White Terriers (WHWTs) diagnosed with aqueous deficient dry eye (ADDE) disease. METHODS: Six ADDE-affected and 13 ADDE-unaffected WHWT dogs were enrolled and underwent clinical assessment and disease scoring, tear osmolarity, phenol red thread test, Schirmer tear test, tear film breakup time, fluorescein staining, Rose bengal and lissamine green vital dye staining, meibometry, corneal esthesiometry, ultrasound pachymetry, optical coherence tomography, in vivo confocal microscopy, and conjunctival biopsy. Subjective assessment of their condition was provided by owner-reported surveys. RESULTS: ADDE-affected WHWT dogs had higher median clinical disease (conjunctiva: 5.75 vs. 0.00; cornea: 14.00 vs. 5.00; total: 17.50 vs. 5.00), vital staining (Rose bengal: 2.25 vs. 1.50; lissamine green: 2.00 vs. 1.00), and histologic disease (conjunctiva: 2 vs. 0) scores when compared with the controls. In addition, ADDE-affected WHWTs had significantly lower phenol red thread test (5.0 vs. 17.5, mm/15 s), Schirmer tear test (3 vs. 20, mm/min), tear film breakup time (3.6 vs. 13.9, s) values and higher area under the curve values for meibometry (394 vs. 245, meibometry units [MU]). There were no significant differences in other tear film tests performed. Advanced imaging revealed decreased tear meniscus height (optical coherence tomography) and variable pigment deposition within corneal epithelial cells (in vivo confocal microscopy). CONCLUSIONS: This comprehensive assessment of ADDE-affected WHWTs depicts the ocular surface changes associated with quantitative lacrimal gland dysfunction. Importantly, ADDE-affected WHWTs may prove a valuable naturally occurring ADDE model for investigating underlying pathophysiological mechanisms and the development of novel therapeutics.

Randomised trial of the clinical utility of an eyelid massage device for the management of meibomian gland dysfunction.

PURPOSE: To compare the single application and two week treatment effects of device-applied (Eyepeace) and manually-applied eyelid massage techniques, as an adjunct to warm compress therapy, on ocular surface and tear film parameters. METHODS: Twenty participants (11 females, 9 males; mean age, 27 ±â€¯11 years) with dry eye symptoms were recruited in a two week, investigator-masked, randomised, contralateral-eye trial. Following 10 min of warm compress therapy application (MGDRx EyeBag®) on both eyes, eyelid massage therapy was applied to one eye (randomised) by device, and to the fellow eye by manual eyelid massage, once daily for 14 days. Ocular surface and tear film measurements were conducted at baseline, and 15 min post-application by a clinician, then again after 14 days of self-administered daily treatment at home. RESULTS: Baseline clinical measurements did not differ between the treatment groups (all p > 0.05). Following two weeks of treatment, tear film lipid layer grade improved significantly with device massage (p = 0.008), and was marginally greater than manual massage by less than 1 grade (p = 0.03). Although immediate post-treatment improvements in tear film stability were observed in both groups (both p < 0.05), no significant long-term cumulative effects or inter-treatment differences in stability measures were detected (all p > 0.05). Visual acuity, tear meniscus height, conjunctival hyperaemia, ocular surface staining, and meibomian gland dropout did not change during the treatment period (all p > 0.05). CONCLUSIONS: Two weeks of treatment with the eyelid massage device, as an adjunct to warm compress therapy, effected marginally greater improvements in tear film lipid layer thickness than the conventional manual technique, which were statistically but not clinically significant. Future parallel group trials with longer treatment periods and a greater range of disease severity are required.

The Effect of Contact Lens Wear on the Cellular Morphology of the Lid Wiper Area.

SIGNIFICANCE: This cross-sectional study presented a link between contact lens wear and changes on the cellular morphology characteristics of the lid wiper (LW) epithelium, which was not visible by LW staining. PURPOSE: The aim of this study was to establish if the duration of contact lens (CL) wear affects the cellular morphology of the LW epithelium. METHODS: This was a cross-sectional study of 100 individuals with different exposures to CL wear: short, moderate, and long experience of CL wear; previous CL wearers; and nonwearers (NWs) as controls. Impression cytology samples were collected from the central upper lid margin (LW area). After fixing, samples were stained with periodic acid-Schiff and haematoxylin for cell morphology analysis and subsequently graded according to the Nelson 0- to 3-point scale. Lid wiper staining was assessed with the aid of lissamine green and graded using the Korb (0- to 3-point) scale. One-way Kruskal-Wallis analysis followed by the Dunn multiple-comparisons test was used for statistical comparison. RESULTS: The Nelson grade for LW epithelium morphology was significantly different between groups (P = .003). Abnormal epithelial morphology as defined by grade 2 or 3 was evident in 66.7% of CL wearers with short experience and 76.5% of CL wearers with moderate experience. This was significantly higher than NWs of whom only 21.5% showed greater than grade 1 (P = .02 and .005, respectively). There was no significant difference between NWs and other groups. Lid wiper staining did not significantly differ between groups (P = .50) or correlate with the Nelson grade (Spearman r = 0.02, P = .08). CONCLUSIONS: Metaplasia of the LW epithelium was significantly greater in the early to moderate stages of CL. This supports the view that mechanical irritation is responsible for LW changes in CL wear. Ceasing CL wear seems to lead to recovery. Lid wiper staining did not reflect the underlying morphological changes.

Development and Evaluation of Semiautomated Quantification of Lissamine Green Staining of the Bulbar Conjunctiva From Digital Images.

Importance: Lissamine green (LG) staining of the conjunctiva is a key biomarker in evaluating ocular surface disease. The disease currently is assessed using relatively coarse subjective scales. Objective assessment would standardize comparisons over time and between clinicians. Objective: To develop a semiautomated, quantitative system to assess lissamine green staining of the bulbar conjunctiva on digital images. Design, Setting, and Participants: Using a standard photography protocol, 35 digital images of the conjunctiva of 11 patients with a diagnosis of dry eye disease based on characteristic signs and symptoms were obtained after topical administration of preservative-free LG, 1%, solution. Images were scored independently by 2 masked ophthalmologists in an academic medical center using the van Bijsterveld and National Eye Institute (NEI) scales. The region of interest was identified by manually marking 7 anatomic landmarks on the images. An objective measure was developed by segmenting the images, forming a vector of key attributes, and then performing a random forest regression. Subjective scores were correlated with the output from a computer algorithm using a cross-validation technique. The ranking of images from least to most staining was compared between the algorithm and the ophthalmologists. The study was conducted from April 26, 2012, through June 2, 2016. Main Outcomes and Measures: Correlation and level of agreement among computerized algorithm scores, van Bijsterveld scale clinical scores, and NEI scale clinical scores. Results: The scores from the automated algorithm correlated well with the mean scores obtained from the gradings of 2 ophthalmologists for the 35 images using the van Bijsterveld scale (Spearman correlation coefficient, rs = 0.79), and moderately with the NEI scale (rs = 0.61) scores. For qualitative ranking of staining, the correlation between the automated algorithm and the 2 ophthalmologists was rs = 0.78 and rs = 0.83. Conclusions and Relevance: The algorithm performed well when evaluating LG staining of the conjunctiva, as evidenced by good correlation with subjective gradings using 2 different grading scales. Future longitudinal studies are needed to assess the responsiveness of the algorithm to change of conjunctival staining over time.

Clinical Effects and Safety of 3% Diquafosol Ophthalmic Solution for Patients With Dry Eye After Cataract Surgery: A Randomized Controlled Trial.

PURPOSE: To compare the efficacies and safety profiles of 3% diquafosol and 0.1% sodium hyaluronate in patients with dry eye after cataract surgery. DESIGN: Randomized controlled trial. METHODS: setting: Soonchunhyang University Hospital, Seoul, South Korea. STUDY POPULATION: In all, 130 eyes of 86 dry eye patients who had undergone cataract surgery between January 2014 and January 2015 were enrolled and randomly divided into a diquafosol group and a sodium hyaluronate group. INTERVENTION: The diquafosol group used diquafosol 6 times a day and the hyaluronate group used sodium hyaluronate 6 times a day after cataract surgery. MAIN OUTCOME MEASURES: Evaluations of efficacy were conducted based on an Ocular Surface Disease Index questionnaire, tear breakup time (TBUT), Schirmer I test, corneal fluorescein and conjunctival lissamine green staining scores, serial measurement of ocular higher-order aberrations (HOAs), corneal HOAs, and uncorrected distance visual acuity test. Safety evaluations were based on anterior chamber inflammation and discontinuation of the eye drops. RESULTS: Objective signs and subjective symptoms were aggravated at 1 week postoperatively and began to recover significantly 4 weeks after surgery. The diquafosol group showed significantly superior TBUT (P < .001), corneal fluorescein (P = .045), and conjunctival staining (P = .001) compared to the sodium hyaluronate group throughout the study period. TBUT (P < .001) and the change in HOAs (P = .018) recovered significantly more quickly in the diquafosol group. The safety evaluations showed no intergroup differences. CONCLUSIONS: Eye drops of 3% diquafosol may be an effective and safe treatment for the management of cataract surgery-induced dry eye aggravation in patients with preexisting dry eye.

Characterization of Upper Eyelid Tarsus and Lid Wiper Dimensions.

OBJECTIVES: To measure various dimensions of the upper tarsal plate and the area of upper lid wiper staining. The repeatability of the method of measurement was investigated. METHODS: Thirty-five healthy non-contact lens wearers were enrolled. The following parameters were measured from digital images of the upper eyelid captured with a slitlamp camera: length, height, and total area of the tarsal plate and area of lid wiper staining (lissamine green). Measurements were performed in a randomized and masked fashion on two separate occasions by the same investigator using ImageJ (National Institutes of Health). Coefficients of repeatability (COR) were calculated. RESULTS: The dimensions (mean±SD) of the tarsal plate were 20.6±1.9 mm length, 7.9±0.8 mm height, and 103.3±18.8 mm total area. The area of lid wiper staining was 2.7±2.0 mm. No association was found between tarsal dimensions and lid wiper staining (all P>0.05). Image analysis COR values were 0.6 mm tarsal length, 0.1 mm tarsal height, 1.2 mm tarsal area, and 0.4 mm lid wiper staining. There was no significant difference between repeated measurements for any parameter (all P>0.05). Limits of agreement were narrow for all parameters, indicating good agreement between repeated measurements. CONCLUSIONS: This work has demonstrated that there is a wide range in the dimensions of the upper tarsal plate in an urban UK population. No association was found between the upper tarsal dimensions and lid wiper staining. ImageJ was shown to be a repeatable method to measure the dimensions of the upper tarsal plate and upper lid wiper staining.

Correlation of Dry Eye Workshop Dry Eye Severity Grading System With Tear Meniscus Measurement by Optical Coherence Tomography and Tear Osmolarity.

OBJECTIVES: To compare tear meniscus measurements obtained by optical coherence tomography (OCT) and tear osmolarity with dry eye severity according to the Dry Eye Workshop (DEWS) classification system. METHODS: Forty-four eyes of 22 patients with dry eye disease (DED) were recruited in this prospective study. In all eyes, ophthalmic examination was performed in the same order as follows: Ocular Surface Disease Index (OSDI) survey, tear film osmolarity measurement with TearLab Osmolarity System, tear meniscus measurements by OCT, corneal fluorescein staining scoring, conjunctival lissamine green staining scoring, tear film breakup time assessment, and anesthetized Schirmer test. Dry eye disease severity was graded according to the DEWS dry eye severity grading system, and the patients were divided into two groups. Group 1 composed of the patients with grades 1 and 2 DED, and group 2 composed of the patients with grades 3 and 4 DED. RESULTS: The mean tear osmolarity value was significantly higher in group 2 (318.9±12.8 mOsm/L) than in group 1 (308.1±8.5 mOsm/L) (P<0.01). The mean tear meniscus height (TMH), tear meniscus depth (TMD), and tear meniscus area (TMA) values were significantly lower in group 2 (172.9±73.5 µm, 121.57±46.2 µm, and 0.013±0.012 mm, respectively) than in group 1 (218.5±70.2 µm, 157.94±49.1 µm, and 0.022±0.013 mm, respectively) (P=0.05, P=0.02, and P=0.026, respectively). There was a negative correlation between TMH and OSDI at the level of 45% (r=-0.450; P<0.05), between TMD and OSDI at the level of 47% (r=-0.470, P<0.05), and lastly between TMA and OSDI at the level of 48.5% (r=-0.485, P<0.05). There was no correlation between OSDI and tear osmolarity (P>0.05). CONCLUSIONS: Both tear osmolarity and tear meniscus OCT measurements comply with the DEWS grading system, and they can be used in the diagnosis and follow-up of dry eye patients in addition to conventional tests.

Comparison of conjunctival staining between lissamine green and yellow filtered fluorescein sodium.

OBJECTIVE: To compare conjunctival staining between lissamine green and fluorescein sodium using a yellow barrier filter. DESIGN: Retrospective cross-sectional study. PARTICIPANTS: Thirteen eyes of 13 patients with dry eyes. METHODS: All patients underwent ocular surface staining with fluorescein sodium and lissamine green. Ocular surface staining with fluorescein sodium was observed through slit-lamp examination under cobalt blue illumination and a yellow filter. Temporal (0-5) and nasal (0-5) conjunctival staining grade was separately graded according to the Oxford scheme. The contrast score was defined as the absolute difference between mean grey values of punctate staining and background. Conjunctival staining grade and contrast scores were compared between these 2 staining methods. RESULTS: The sum of the conjunctival staining grade (0-10) for the nasal and temporal conjunctiva was 4.7 ± 2.5 for fluorescein sodium, which was significantly higher than that of lissamine green, 4.1 ± 2.5 (p = 0.005). The contrast score of temporal and nasal conjunctival staining was 34.6 ± 12.0 and 34.7 ± 10.3, respectively, for fluorescein sodium, which was also significantly greater than for lissamine green (23.4 ± 8.1 and 21.0 ± 9.0, respectively; p = 0.003 and p = 0.003, respectively). CONCLUSIONS: Conjunctival staining with fluorescein sodium and a yellow filter seems to be more sensitive for the detection of conjunctival damage than lissamine green. Fluorescein staining with the yellow filter has the advantage of simultaneous observation of both corneal and conjunctival damage in patients with dry eye without the need for additional vital staining.

Análise quantitativa e qualitativa do filme lacrimal nos pacientes submetidos a PRK e LASIK com femtossegundo / Analysis quantitative and qualitative of the tear film in patients undergoing PRK and LASIK with femtosecond

Objetivo: Analisar a secreção lacrimal, coloração da superfície ocular e estabilidade do filme lacrimal em indivíduos submetidos à cirurgia de PRK e LASIK com laser de femtossegundo (femto LASIK). Métodos: Vinte olhos de 10 pacientes submetidos à técnica de Femto LASIK e 11 olhos de 6 pacientes submetidos à técnica de PRK foram estudados de forma prospectiva, longitudinal e intervencionista. Tempo de rotura do filme lacrimal (TRFL), teste de Schirmer basal e coloração da superfície ocular com lissamina verde foram analisados no pré-operatório (pré), no 15º e no 30º dia pós-operatório (15º pós e 30º pós, respectivamente). Resultados: Agrupando todos os olhos, observou-se que o TRFL reduziu-se de forma estatisticamente significante no 15º pós em relação ao valor pré-operatório (p=0,025), mantendo-se reduzido no 30º pós (p= 0,001); não houve diferença estatisticamente significativa entre o 15º pós e o 30º pós (p=0,219). No teste da lissamina verde, houve aumento significativo desse escore, no 15º pós em relação ao período pré-operatório (p=0,021), havendo, posteriormente, redução no 30º pós (p=0,010). No teste de Schirmer basal, não foi detectada mudança estatisticamente significante ao longo dos três momentos (p=0,107). Comparando-se os testes TRFL, lissamina verde ou Schirmer basal, nos dois grupos estudados (PRK e LASIK), não houve diferença estatisticamente significante em nenhum dos três momentos (pré, 15º pós e 30º pós). Conclusão: Evidenciou-se alteração do filme lacrimal nos pacientes submetidos à cirurgia refrativa, quando foram utilizados os testes de TRFL e lissamina verde. Nas duas técnicas empregadas, não houve diferença estatisticamente significante de alteração do filme lacrimal, quando comparadas entre si (PRK e LASIK). .

Purpose: To evaluate tear film stability, ocular surface staining and tear secretion in patients undergoing PRK and femtosecond laser LASIK. Methods: Twenty eyes of 10 patients submitted to femtosecond laser LASIK and 11 eyes of the 6 patients submitted do PRK underwent tear film break-up time (TBUT), Schirmer’s basal and lissamine green staining measurements pre and postoperatively on days 15 (PO 15) and 30 (PO 30). Results: When grouping all eyes TBUT was reduced on PO 15 (p=0.025) and on PO 30 (p=0.001) compared to preoperative values. No difference was found between PO 15 and PO 30 (p=0.219). Compared to preoperative measurements, lissamine green test demonstrated a significant increase in score on PO 15 (p=0.021) and a significant reduction on PO 30 (p=0.010), when both groups were analyzed together (all 42 eyes). No changes in Schirmer’s basal test were detected in all 3 time periods (p=0.107). TBUT, lissamine green and Schirmer’s basal measurements were no different in all 3 time periods when both groups (PRK and femtosecond laser LASIK) were compared. Conclusion: TBUT and lissamine green measurements were altered after refractive surgery regardless the technique (PRK or femtosecond laser LASIK). When comparing one technique to the other, no difference was found in all measurements. .

Optimizing evaluation of Lissamine Green parameters for ocular surface staining.

PURPOSE: The recently published seminal dry eye workshop proceedings defined Lissamine Green (LG), an organic dye, as a gold standard for demonstrating ocular surface staining. The purpose of the current study was to determine the optimal parameters of 1% LG instillation for the ocular surface examination in dry eye patients. DESIGN: Prospective and observational quality improvement study. METHODS: A quality improvement study evaluated 16 eyes from eight dry eye patients with different levels of severity. LG (1%), in three volumes (5, 10, and 20 µl) was instilled into the conjunctival cul-de-sac, and four masked observers with different levels of clinical expertise examined the patients with and without red filter. The staining pattern of the conjunctiva and cornea was documented with the Oxford scale within 4 min of LG instillation. Optimal volume and inter-observer reliability were assessed. RESULTS: All dye volumes were tolerated well by all patients. Experienced observers preferred 10 µl volume because of the ease of examination and accuracy. Although instillation of 20 µl yielded similar scores as 10 µl, it resulted in overflow of the lid and facial skin staining. The use of red filter significantly improved reading scores (P<0.01). Inter-observer reliability was higher for conjunctival scores than for corneal scores for all patients. The highest reliability was demonstrated with 10 µl volume and increased with greater experience of the observer. CONCLUSIONS: Ocular surface examination with instillation of 10 µl 1% LG has good inter-observer reliability and is well tolerated. Observation through a red filter facilitates the examination.

An evaluation of the efficacy of fluorescein, rose bengal, lissamine green, and a new dye mixture for ocular surface staining.

PURPOSE: Sodium fluorescein is considered the premier dye for corneal staining and, similarly, rose bengal (RB) for conjunctival staining. A mixture of 1% fluorescein and 1% rose bengal has been reported as advantageous in daily practice. Mixtures of lissamine green with other ocular stains have not been reported. The purposes of this study were to review the clinical staining characteristics of fluorescein, rose bengal, and lissamine green in controlled dose and concentration and determine whether optimal staining of the cornea and bulbar conjunctiva are possible by using dye mixtures. METHODS: Sixteen 10-microL solutions of fluorescein, rose bengal, lissamine green, and their mixtures were evaluated. Fourteen subjects with a diagnosis of dry eye were tested for staining with various combinations of the dyes. Examination of staining was made by using standard clinical practices. RESULTS: A mixture of 2% fluorescein and 1% rose bengal was the most efficacious staining mixture for the cornea and conjunctiva, but moderate to marked discomfort was reported. The mixture of 2% fluorescein and 1% lissamine green did not result in discomfort and provided optimal corneal and conjunctival staining with only slightly less efficacy than 2% fluorescein and 1% rose bengal; 2% and 3% lissamine green produced burning and discomfort. The fluorescent characteristics of fluorescein were not significantly altered by the addition of 1% lissamine green. The preferred mixture for simultaneous and efficacious staining of the cornea and conjunctiva without an adverse sensation was 2% fluorescein and 1% lissamine green. CONCLUSIONS: A mixture of 2% fluorescein and 1% lissamine green offers excellent simultaneous corneal and bulbar conjunctival staining and could replace the use of individual dyes for ocular staining and contact lens practice.

Pattern of vital staining and its correlation with aqueous tear deficiency and meibomian gland dropout.

PURPOSE: To assess the relationship between the degree and anatomic location of ocular surface vital staining and the degree of aqueous tear deficiency in patients with dry eye, as measured by the Schirmer test, and the presence of meibomian gland dropout. METHODS: Twenty-two patients with dry eye (nine men and 13 women; mean age, 60 years) were prospectively enrolled in the study. Eleven subjects with no ocular disease were considered control subjects. Lissamine green vital staining, Schirmer test, and meibomian gland dropout were evaluated. RESULTS: Patients were stratified based on their vital staining pattern (presence or absence of corneal staining). A progressive decrease in the Schirmer test result was found when the vital staining score increased (r = -0.655; P<0.0001). Patients with dry eye showed an increase in meibomian gland dropout when compared to control subjects (P

[Substances for staining biological tissues: use of dyes in ophthalmology]. / Substanzen zur Anfärbung von biologischem Gewebe -- Farbstoffe in der Ophthalmologie.

The azo dye trypan blue and the cyanine dye indocyanine green have been used in ophthalmology since the 1980 s to dye the anterior capsule in cataract operations and to stain epiretinal membranes in order to remove the membrana limitans interna (ILM). By means of a standardized in vitro test in accordance with DIN EN ISO 10993, it has now been shown that both dyes and the theoretically possible alternatives - the anthrachinone alizarincyanine green and the trityl dyes fast green and lissamine green - are cytotoxic in the normal concentrations of maximum 1.5 g/l. On the other hand, the new product Blueron(R) with the trityl dye patent blue, which has been developed to dye the anterior capsule, is completely biocompatible, even at a high dosage of up to 2.8 g/l.

A quantitative assessment of the location and width of Marx's line along the marginal zone of the human eyelid.

PURPOSE: To quantitatively assess the staining of Marx's line with lissamine green dye. METHODS Forty white (European) subjects, aged 18 to 78 years, were assessed. The subjects had no major eye disease, although some had mild to moderate blepharitis. Lissamine green dye strips were used to stain the marginal zone of the palpebral conjunctiva, and high-magnification photo slit lamp images were taken of the everted upper eyelid along with a scale rule. From prints at 1600x, the distances between the main tarsal (Meibomian) gland orifices, the width of the lissamine green-stained Marx's line, and the location of its anterior border were measured. RESULTS: The group-mean distance between the main tarsal gland orifices was 0.81 +/- 0 16 mm, whereas the width of the lissamine green-stained line was 0.10 +/- 0.09 mm. In most subjects, the lissamine green-stained line was posterior to the tarsal gland orifices, and the location of the proximal (anterior) edge of this line averaged +0.13 mm from a reference line drawn through the orifices. CONCLUSION: Lissamine green dye highlights a very distinct line of cells along the marginal zone. This line of cells is narrow and is thought to be the natural site of frictional contact between the eyelid margin and the surfaces of the bulbar conjunctiva and cornea, rather than the edge of the tear meniscus or location of the edge of the lacrimal river.

The use of vital dyes in corneal disease.

Vital dyes such as fluorescein, rose bengal, and lissamine green B, among others, have been an important tool in clinical ophthalmology in diagnosis and evaluation of various ocular surface disorders. Each dye, having a unique chemical structure, has distinct characteristics that allows its preferential use in different ocular pathologies. In this article, history of vital dyes and the use of popular current vital dyes in corneal disease are reviewed.