Carbon Black Tinted Contact Lenses for Reduction of Photophobia in Cystinosis Patients.

PURPOSE: To examine if current development on using contact lenses for drug delivery of cysteamine to treat ocular symptoms of cystinosis can be tinted to mitigate photophobia common in patients by reducing transmittance Methods: Commercial contact lenses were placed in a carbon black solution to examine loading after lens synthesis. Silicone hydrogel contact lenses were also synthesized with carbon black added prior to UV curing. Transmittance was measured using UV-vis spectrophotometry over the range of 190-1190 nm and compared to unmodified contact lenses. Lens parameters of refractive index, ion permeability, and Young's modulus were measured using a refractometer, release of sodium chloride, and the cantilever method. Cysteamine release was measured by loading lenses into 5% cysteamine solution and then monitoring the release of the drug using UV-vis spectrophotometry. Vitamin E diffusion barriers were also added to lenses via ethanol solution, and the release of cysteamine from these modified lenses was also examined. RESULTS: No leeching of carbon black was detected during experiments. Loading of pre-made contact lenses led to uneven distribution of carbon black throughout lens. Adding 0.3% carbon black to lens monomer solution prior to UV-curing led to even distribution and a transmittance reduction of approximately 50%. Ion permeability was reduced from 6.19 ± 0.90 x 10-3 to 1.28 ± 0.06 x 10-3 mm2 min-1, and Young's modulus was decreased from 1.58 ± 0.08 to 1.29 ± 0.06 MPa. Cysteamine releases from carbon black lenses with and without vitamin E were comparable to controls, although the loading solution of vitamin E/ethanol had to be tripled to achieve a similar mass loading to control. CONCLUSIONS: Carbon black increases the softness of contact lenses, but a loading of 0.3% maintains lens parameters required for wear. The release of cysteamine is also possible with carbon black lenses, albeit requiring a higher loading concentration of vitamin E to achieve similar release times.

Spectroscopy of Oxygen-Sensitive Material for Measuring Contact Lens Oxygen Transmissibility.

PURPOSE: Oxygen permeability or transmissibility is a crucial parameter for contact lenses to ensure that extended wear will not induce corneal hypoxia. This work tests a new method of using the oxidation of cysteamine, an oxygen-sensitive chemical, to quantify the oxygen transmissibility of current commercial contact lenses and contact lenses loaded with vitamin E. METHODS: 3D printing was used to modify eye drop bottles and quartz cuvettes to create systems that allowed insertion of a contact lens in between the cysteamine solution and air. Both systems were exposed to atmospheric conditions where the only path of entry for oxygen was through the contact lens. The entering oxygen reacted with cysteamine, and the rate of cysteamine oxidation was measured using UV-vis spectrophotometry. The rate was then stoichiometrically related to oxygen transmissibility. RESULTS: The eye drop method predicted transmissibility values within 9% of established, commercial values. The cuvette method predicted values within 10% of established values for silicone hydrogel lenses without any correction factor and within 11% for poly-hydroxyethyl-methacrylate lenses after correcting for oxygen entering the system. Incorporation of 20% (w/w) vitamin E into Acuvue® Oasys® lenses did not have a significant impact on the oxygen transmissibility. CONCLUSIONS: Both methods presented in this work can reliably measure oxygen transmissibility of contacts lenses or other materials. Further improvements in manufacturing could lead to improved accuracy and reliability, allowing wider use of this method for quantifying the oxygen transport in contact lenses.

Pigmented contact lenses for managing ocular disorders.

Blocking a selected wavelength range from the light spectrum can have multiple benefits. Ultra-violet (UV) radiation is detrimental to the retina, necessitating its blocking through sunglasses and contact lenses. The near-visible light also has enough energy to cause damage but, is typically not blocked by commercial lenses. Filtering light can also be useful to patients with migraines, amblyopia, and color blindness. Here, to achieve blocking, incorporation of pigments extracted from colored agro-products into contact lenses is explored. Pigment extraction from food powders including turmeric, spinach, paprika, and woad powders in ethanol is demonstrated. Lens immersion in pigment concentrated ethanol is done to facilitate swelling, allowing rapid pigment uptake. Pigment incorporation ensures the absence of visible light scattering, lens opacity, and leaching. The characterization of pigmented lenses is done through absorptivity and transmittance measurements. Degradation measurements investigate the stability of the green pigment extract from spinach powder with time. p-HEMA and silicone hydrogels loaded with >400 µg/g turmeric pigment act as class 1 UV blockers retaining >90% visible light transparency and screening >95% of the UVR spectra. Spinach, paprika, and woad powder loaded silicone lenses mitigate >20% visible light transmission from selective wavelengths finding applications in photophobia, amblyopia treatment, and color vision deficiency management.

Correction to: Controlled delivery of pirfenidone through vitamin E-loaded contact lens ameliorates corneal inflammation.

In the original article the typesetter made several errors. Figures 7 and 9 are incorrect. Following are the correct figures.

Potential role of stromal collagen in cystine crystallization in cystinosis patients.

Cystinosis is a genetic disease that leads to the accumulation of intracellular cystine crystals in all organs including cornea due to the loss of cystine efflux transporters in the lysosome of the cells. While the mechanism for formation of intracellular cystine is well understood for most organs, it does not explain many observations for crystal accumulation in the cornea of cystinosis patients. First, the crystals in cornea are extracellular and needle-like with several hundred microns length which is in sharp contrast with the rectangular or hexagonal crystals found throughout other organs. Second, these crystals are arranged parallel to the stromal collagen, which is a unique to the cornea. Third, crystal growth in the cornea reaches a saturation point after where no further crystallization occurs. We propose a hypothesis supported by in vitro and ex vivo data to explain these observations. We hypothesize that the stroma crystals form extracellularly due to the ionic interactions between the cystine diffusing into the eye and collagen fibrils present in the stroma. We examine cystine crystal growth both with in vitro polymer solutions and ex vivo in rabbit cadaver eyes to show that negatively charged polymers lead to the formation of more cystine precipitation in aqueous solution and that needle-like cystine crystal formation can be observed only in presence of certain polyelectrolytes including collagen. This proposed mechanism explains many of the yet unanswered questions but it needs further support from in vivo studies. The improved understanding could lead to improved treatment of corneal cystinosis.

Novel approaches for improving stability of cysteamine formulations.

Cystinosis is a genetic disorder that leads to the formation of cystine crystals in many organs in the body including cornea. Ocular manifestation of this disease is treated by eye drops of cysteamine which can easily oxidize into its disulfide cystamine. The rapid oxidation limits the shelf life as well the duration during which the drug can be used after opening the eye drop bottle. We evaluate two approaches of preventing the oxidation of cysteamine with the goal of increasing the time of use after opening the bottle to one month. The first approach integrates antioxidants such as catalase enzyme and vitamins C and E into the aqueous solution. Results show that catalase is the most effective additive as it decreases the oxidation rate by 58%, which on its own is not sufficient to reach targeted one month stability. The second approach focuses on incorporating diffusion barriers to prevent oxygen from reaching the cysteamine solution. This was accomplished by two methods: formulation of a hydrophobic layer which floats on the surface of the aqueous solution and integration of OMAC® oxygen-resistant material into the eye drop bottle. Both methods delay the onset of cysteamine degradation and decrease the rate of degradation. In particular, an eye drop bottle with three layers of OMAC® has less than 10% degradation after one month of opening the bottle and withdrawing a drop each day. By integrating all three methods, we designed a system where >90% of cysteamine remains in the active form for 70 days after opening the bottle. In addition, we examine the use of OMAC® as heat-sealed pouches for storage of cysteamine eye drop bottles during packaging to eliminate the need for the current approach of freezing the formulation during shipping. The results show that such heat-sealed pouches would keep cysteamine stable for over one year at ambient conditions.

Controlled delivery of pirfenidone through vitamin E-loaded contact lens ameliorates corneal inflammation.

Chemical injury by alkali burn is a major cause of corneal blindness in the clinical setting. Current management advocates multiple therapies aimed to prevent inflammation, initiate quick re-epithelialization, arrest the fibrosis, and avoid dry eye and pain by using bandage contact lenses. We hypothesized sustained delivery of the anti-inflammatory, antifibrotic drug pirfenidone through vitamin E-loaded contact lenses as a logical single approach to counter the pathology involved. Vitamin E particles were created in situ in commercial silicon hydrogel contact lenses by soaking the lenses in a vitamin E-ethanol solution. The vitamin E-laden lenses were then placed into pirfenidone-saline solution to load the drug into the lens. The contact lenses were evaluated by both in vitro and in vivo means. For in vitro, lenses were placed into 3 mL of saline solution. The concentration of pirfenidone released was measured by UV-vis spectrophotometry. The contact lenses were implanted in rabbit eyes following the alkali burn; the drug availability in the aqueous humor was evaluated by HPLC at various time points 10 min, 30 min, 2 h, and 3 h; and gene expression of inflammatory cytokines IL-1ß, TNF-α, and TGF-ß1 was evaluated in the cornea at the end of the study period. In another group of rabbits inflicted with alkali injury, the corneas were graded after 7 days of contact lens implantation with and without pirfenidone. A mathematical model was developed for delivery of the drug to the cornea and aqueous humor after a contact lens is inserted in the eye. The model was validated with experimental data and used to determine the bioavailability both for contact lenses and eye drops. In vitro release of unmodified commercial contact lenses saw a release time of approximately 20 min, with a partition coefficient of 2.68 ± 0.06. The release of pirfenidone from 20% vitamin E-loaded lenses saw a release time of approximately 80 min, with a partition coefficient of 4.20 ± 0.04. In vivo, the drug was available in the aqueous humor for up to 3 h. Gene expression of inflammatory cytokine IL-ß1 and profibrotic growth factor TGF-ß1 was significantly suppressed in corneas treated with pirfenidone contact lenses. A week after the alkali burn, the eyes with pirfenidone contact lenses showed significant improvement in corneal haze in comparison to the control eyes. About 50% of the drug loaded in the lens reached the aqueous humor compared to 1.3% with eye drops. Vitamin E-loaded contact lenses serve as a suitable platform for delivery of pirfenidone following alkali burn in rabbit eyes; positive pre-clinical outcome identifies it as promising therapy for addressing corneal inflammation and fibrosis. The bioavailability is about 40-fold higher for contact lenses compared to that for eye drops.

In vitro drug release and in vivo safety of vitamin E and cysteamine loaded contact lenses.

Cystinosis is an orphan disease caused by a genetic mutation that leads to deposition of cystine crystals in many organs including cornea. Ophthalmic manifestation of the disease can be treated with hourly instillation of cysteamine eye drops. The hourly eye drop instillation is tedious to the patients leading to poor compliance and additionally, significant degradation of the drug occurs within one week of opening the bottle, which further complicates this delivery approach. This paper focuses on designing a contact lens to treat the disease with improved efficacy compared to eye drops, and also exploring safety of the drug eluding contact lens in an animal model. Our goal is to design a lens that is safe and that can deliver a daily therapeutic dose of cysteamine to the cornea while retaining drug stability. We show that cysteamine diffuses out rapidly from all lenses due to its small size. Vitamin E incorporation increases the release duration of both ACUVUE®OASYS® and ACUVUE® TruEyeTM but the effect is more pronounced in TruEyeTM likely due to the low solubility of vitamin E in the lens matrix and higher aspect ratio of the barriers. The barriers are not effective in hydrogel lenses, which along with the high aspect ratio in silicone hydrogels suggests that barriers could be forming at the interface of the silicone and hydrogel phases. The presence of vitamin E has an additional beneficial effect of reduction in the oxidation rates, likely due to a transport barrier between the oxygen diffusing through the silicone channels and drug located in the hydrogel phase. Based on this study, both Acuvue®OASYS® and ACUVUE® TruEyeTM can be loaded with vitamin E to design a cysteamine eluting contact lenses for effective therapy of cystinosis. The lenses must be worn for about 4-5 hr. each day, which is less than the typical duration of daily-wear. The vitamin E and cysteamine loaded lenses did not exhibit any toxicity in a rabbit model over a period of 7-days.

Incorporation of drug particles for extended release of Cyclosporine A from poly-hydroxyethyl methacrylate hydrogels.

Cyclosporine A is prescribed for a number of ophthalmic applications such as dry eyes, uveitis in children and adolescents, vernal keratoconjunctivitis, and peripheral ulcerative keratitis. Extended release of cyclosporine from contact lenses has been explored due to the significant benefits of increased bioavailability in comparison with eye drops. Incorporation of drug loaded particles is considered to be a promising approach for increasing the drug release duration. Here we explore the feasibility of extended release of cyclosporine and possibly other hydrophobic drugs by dispersing particles that are 100% drug rather than drug loaded particles. The expected benefits are high drug loading and extended release. Specifically, we explore transport of cyclosporine in hydroxyethyl methacrylate gels for the case when the gel is loaded with high concentration of drug leading to in situ formation of particles. We explore whether we can increase the release duration from the gels by incorporation of the particles, without sacrificing light transmission which is a critical property for contact lenses. Hydrogels were prepared by free radical UV initiated polymerization with drug dissolved in the monomer solution at varying loadings. Drug release kinetics were measured from the particle loaded lenses and fitted to the Higuchi model to determine the diffusivity. The measured diffusivity is two orders of magnitude lower than estimates from Brinkman model. The differences were attributed to the high partition coefficient of about 150, which implies that a majority of the drug in the gel is bound to the polymer. The bound drug can diffuse along the surface or desorb and diffuse. The diffusivity estimates match the measured values after binding is taken into consideration. Light transmittance was measured to determine whether particle incorporation reduces the transparency. Results showed that the drug release duration could be controlled by increasing the drug loading but the transmittance was significantly reduce particularly at high drug loadings, which suggest that this approach may have limited applicability for contact lenses, but could be useful in other applications where light transmission is not critical.

Effect of the surface layer on drug release from delefilcon-A (Dailies Total1®) contact lenses.

Contact lenses are receiving significant attention for delivering ophthalmic drugs with higher bioavailability compared to eye drops. Here we explore drug transport from delefilcon-A Dailies Total1® lenses which are designed to have a thin, high-water content layer on the surface. Our goal is to determine the impact of this high water content layer on drug transport for both hydrophobic (dexamethasone and cyclosporine) and hydrophilic (timolol and levofloxacin) drugs. Drugs were loaded into the lens by soaking in aqueous drug solutions till equilibrium, followed by release in phosphate buffered saline. The concentration data during release was fitted to the diffusion equation without considering the surface layer. If fits were poor, the surface layer was include in the model, as a burst release. Results showed that surface layer resulted in a burst release of about 35% of the loaded drug for the two hydrophilic drugs, and the model did not fit the data unless the surface layer was incorporated as a burst release. For the hydrophobic drugs, there was no burst release and the model fitted the data without including the surface layer likely because of the low partition coefficient of the hydrophobic drugs in the surface layer compared to the lens. The results further confirm the presence of the high water content surface layer on the Dailies Total1® lenses. The release profile of the burst release for hydrophilic drugs could be therapeutically useful for antibiotics where a high dose is desirable initially. The effect of vitamin E loading-an established procedure for increasing drug release time in other commercial lenses, was also tested on the release of timolol maleate and levofloxacin. A 20% vitamin E loading was found to increase the release time of timolol and levofloxacin by a factor of 5 and 3-fold respectively, but this increase proved much less effective compared to vitamin E's effect on other commercial silicone hydrogels.

Emergency Physician-Performed Bedside Ultrasound in the Evaluation of Acute Appendicitis in a Pediatric Population.

Background/Objective Many pediatric emergency departments in the United States have adopted a staged ultrasound and CT pathway for the diagnosis of acute appendicitis. However, most algorithms only include radiology-performed ultrasound (RUS) and not emergency physician- performed bedside ultrasound (BUS). Our objective was to determine if emergency physician-performed BUS provides sufficient diagnostic accuracy for acute appendicitis in a pediatric population, thereby limiting additional cost and/or delays in disposition. Methods This is a single-center prospective study of pediatric patients with concern for and requiring further work-up for acute appendicitis. Each patient had a focused bedside ultrasound (BUS) performed by an emergency physician with training in BUS. Diagnostic accuracy was compared with surgical pathology standard, as well as radiology- performed ultrasound (RUS), computed tomography (CT), and clinical follow-up. Results Among46 enrolledpatients, 12were diagnosed with acute appendicitis (26%). There were no negative laparotomies in those who had surgery. There was one case of missed appendicitis at 4-week follow-up. BUS had a sensitivity of 100% (95% Cl: 72% to 100%) and. a specificity of 81% (61% to 93%) when the app6ndix'was visualized (37). This resulted in positive likelihood ratio of5.2 and a negative likelihood ratio ofo. In the cases where the appendix was not visualized on BUS (9), 1 patient was diagnosed with appendicitis, and the other 8 patients were negative for appendicitis. In RUS both the sensitivity and specificity was 100% when the appendix was visualized. The sensitivity and specificity of CT in our studywas 90% and 100% respectively. Conclusions Emergency physicians can perform bedside ultrasound with high accuracy for acute appendicitis in a pediatric population. When the appendix is not visualized by ultrasound, a staged ultrasound and CT pathway should be considered.

Comparisons of prediction equations for estimating energy expenditure in youth.

OBJECTIVES: The purpose of this study was to compare the validity of Actigraph 2-regression models (2RM) and 1-regression models (1RM) for estimation of EE in children. DESIGN: The study used a cross-sectional design with criterion estimates from a metabolic cart. METHODS: A total of 59 children (7-13yrs) performed 12 activities (randomly selected from a set of 24 activities) for 5min each, while being concurrently measured with an Actigraph GT3X and indirect calorimetry. METRMR (MET considering one's resting metabolic rate) for the GT3X was estimated applying 2RM with vector magnitude (VM2RM) and vertical axis (VA2RM), and four standard 1RMs. The validity of the 2RMs and 1RMs was evaluated using 95% equivalence testing and mean absolute percent error (MAPE). RESULTS: For the group-level comparison, equivalence testing revealed that the 90% confidence intervals for all 2RMs and 1RMs were outside of the equivalence zone (range: 3.63, 4.43) for indirect calorimetry. When comparing the individual activities, VM2RM produced smaller MAPEs (range: 14.5-45.3%) than VA2RM (range, 15.5-58.1%) and 1RMs (range, 14.5-75.1%) for most of the light and moderate activities. CONCLUSIONS: None of the 2RMs and 1RMs were equivalent to indirect calorimetry. The 2RMs showed smaller individual-level errors than the 1RMs.

Mechanistic modeling of ophthalmic drug delivery to the anterior chamber by eye drops and contact lenses.

Ophthalmic drug for the anterior chamber diseases are delivered into tears by either eye drops or by extended release devices placed in the eyes. The instilled drug exits the eye through various routes including tear drainage into the nose through the canaliculi and transport across various ocular membranes. Understanding the mechanisms relevant to each route can be useful in predicting the dependency of ocular bioavailability on various formulation parameters, such as drug concentration, salinity, viscosity, etc. Mathematical modeling has been developed for each of the routes and validated by comparison with experiments. The individual models can be combined into a system model to predict the fraction of the instilled drug that reaches the target. This review summarizes the individual models for the transport of drugs across the cornea and conjunctiva and the canaliculi tear drainage. It also summarizes the combined tear dynamics model that can predict the ocular bioavailability of drugs instilled as eye drops. The predictions from the individual models and the combined model are in good agreement with experimental data. Both experiments and models predict that the corneal bioavailability for drugs delivered through eye drops is less than 5% due to the small area of the cornea in comparison to the conjunctiva, and the rapid clearance of the instilled solution by tear drainage. A contact lens is a natural choice for delivering drugs to the cornea due to the placement of the contact in the immediate vicinity of the cornea. The drug released by the contact towards the cornea surface is trapped in the post lens tear film for extended duration of at least 30min allowing transport of a large portion into the cornea. The model predictions backed by in vivo animal and clinical data show that the bioavailability increases to about 50% with contact lenses. This realization has encouraged considerable research towards delivering ocular drugs by contact lenses. Commercial contacts are, however, not ideal for drug delivery due to the short release durations which may necessitate wearing multiple lenses each day, reducing the viability of this approach. Recent research has focused on designing contacts that retain all critical properties while increasing the release durations to a few hours or a few days. Beagle dog studies with contact lenses containing vitamin E nanobarriers to attenuate drug transport have shown promising results. Human studies using contacts for drug delivery have also been conducted for allergy therapy but drug eluting contacts are not available in the market for any therapy.

Therapeutic contact lenses: a patent review.

INTRODUCTION: Ophthalmic drugs are almost exclusively delivered via eye drops in spite of several deficiencies, including low bioavailability and poor compliance. Contact lenses have the potential to increase bioavailability by an order of magnitude, while also improving compliance. AREAS COVERED: In this review, the authors summarize advances in therapeutic contact lenses. The major focus of the review is on patents on drug-eluting contact lenses, but non-drug-eluting contacts that offer a therapeutic benefit are also included. The content is divided based on the broad technology of the patent, including novel materials, molecular imprinting, diffusion barriers, colloid encapsulation, surface modification, layering, and other novel approaches. In addition to patents, research publications are also included to facilitate the understanding of the mechanisms and challenges. EXPERT OPINION: Among all non-invasive alternatives, contact lenses offer the highest bioavailability to the cornea due to the location of the lens in the immediate vicinity of the cornea. Several approaches have been patented to improve contact lens design for an extended release duration of drugs. Many technologies have successfully integrated suitable drug release profiles into contact lenses, but drug-eluting contacts are not yet commercialized likely due to regulatory challenges, including the high costs of clinical trials.

Quantifying Loss Caused by Ray Blight Disease in Tasmanian Pyrethrum Fields.

The efficacy of newly implemented fungicide recommendations on reducing the intensity of ray blight disease caused by Phoma ligulicola to achieve site-specific attainable yield potentials in Tasmanian pyrethrum fields was quantified over two seasons in 46 and 51 fields during the 2003 and 2004 growing seasons, respectively. Disease intensity and yield in two plots (10 × 24 m), one following the commercial fungicide protocol recommendations and the second receiving no fungicide, were assessed in each pyrethrum field. The commercial fungicide protocol consisted of one application of azoxystrobin at 150 g a.i./ha, followed by two applications of a tank mixture of difenoconazole at 125 g a.i./ha and chlorothalonil at 1,008 liters a.i./ha at 14- to 21-day intervals. This program resulted in significant decreases in defoliation severity and the incidence of stems and flowers with ray blight, and increases in the height of stems and number of flowers produced per stem in October and November. In plots receiving the commercial fungicide protocol, the dry weight of flowers was increased by 76 and 68% in 2003 and 2004, respectively. Moreover, pyrethrin yield increased by 81 and 78% when the commercial fungicide protocol was used compared with the nontreated plots. Tobit regression was used to examine the relationships and thresholds among disease intensity measures (defoliation severity, stem severity, and incidence of flowers with ray blight) assessed just prior to harvest. This regression utilized a left-censored regression model to define subminimal thresholds, as none of the disease intensity measures could be less than 0. Defoliation severity had a threshold of 35.3% before stem severity linearly increased and a threshold of 38.2% before the incidence of flowers with ray blight linearly increased. Finally, the threshold for stem severity was 13.7% before the incidence of flowers with ray blight linearly increased. These thresholds can be used to assist growers in making disease management decisions with the objective of minimizing loss of flowers by maintaining defoliation severity below the critical point at which the incidence of flowers with ray blight begins to linearly increase.

Mining EST databases to resolve evolutionary events in major crop species.

Using plant EST collections, we obtained 1392 potential gene duplicates across 8 plant species: Zea mays, Oryza sativa, Sorghum bicolor, Hordeum vulgare, Solanum tuberosum, Lycopersicon esculentum, Medicago truncatula, and Glycine max. We estimated the synonymous and nonsynonymous distances between each gene pair and identified two to three mixtures of normal distributions corresponding to one to three rounds of genome duplication in each species. Within the Poaceae, we found a conserved duplication event among all four species that occurred approximately 50-60 million years ago (Mya); an event that probably occurred before the major radiation of the grasses. In the Solanaceae, we found evidence for a conserved duplication event approximately 50-52 Mya. A duplication in soybean occurred approximately 44 Mya and a duplication in Medicago about 58 Mya. Comparing synonymous and nonsynonymous distances allowed us to determine that most duplicate gene pairs are under purifying, negative selection. We calculated Pearson's correlation coefficients to provide us with a measure of how gene expression patterns have changed between duplicate pairs, and compared this across evolutionary distances. This analysis showed that some duplicates seemed to retain expression patterns between pairs, whereas others showed uncorrelated expression.