Artificial Tear Fluid Research Articles

Towards the real-time monitoring of glucose in tear fluid: Holographic glucose sensors with reduced interference from lactate and pH

Glucose-selective holographic sensors were fabricated from unique tetrahedral 2-acrylamidophenylboronic acid (2-APB) incorporated with co-monomers poly(ethylene glycol) acrylate (PEG), (3-acrylamidopropyl)trimethylammonium chloride (ATMA) and [2-(acryloyloxy)ethyl]-trimethylammonium chloride (AETA) into thin hydrogel films which were transformed into volume holograms using a diffusion method coupled with holographic recording using a frequency-doubled Nd:YAG laser (532 nm). The results showed that the 2-APB-based holographic sensors contracted upon addition of glucose due to the formation of a 2:1 complex between the tetrahedral 2-APB and glucose. More significantly, the 2-APB-based holographic sensors had greatly reduced lactate dependence and a hugely reduced pH effect over the physiological range of pH. These features are vital for development of contact lens-based glucose sensor, where the pH variability is greater (pH 5.8–7.8) and the lactate concentration is substantially higher than in blood. Furthermore, the 2-APB-based holographic sensors also displayed fast response to glucose. The successful union of holograms and the tetrahedral 2-APB receptor for glucose detection in artificial tear fluid is also demonstrated. This new type of holographic sensors responding to glucose with features of minor pH effect and negligible interference from lactate is applicable to the detection of glucose concentrations in tear fluid for the management of diabetes.

Fast Responsive Crystalline Colloidal Array Photonic Crystal Glucose Sensors

We developed new photonic crystal polymerized crystalline colloidal array (PCCA) glucose sensing materials, which operate on the basis of formation of cross-links in the hydrogel. These materials are composed of hydrogels that embed an array of approximately 100-nm-diameter monodisperse polystyrene colloids that Bragg diffract light in the visible spectral region. The hydrogels change volume as the glucose concentration varies. This changes the lattice spacing, which changes the wavelength of the diffracted light. In contrast to our previous glucose sensing photonic crystal materials, we no longer require Na+ chelating agents. These photonic crystal materials are being designed for use in glucose sensing contact lens for people with diabetes mellitus. We describe methods to speed up the response kinetics of these PCCA sensing materials. Rapid-response kinetics is achieved by controlling the elasticity and the hydrophilic-hydrophobic balance of the hydrogel system. A more hydrophobic hydrogel composition is obtained by copolymerizing n-hexylacrylate into an acrylamide-bisacrylamide hydrogel. The response rate significantly increases to where it fully responds within 90 s to the average glucose concentrations found in blood (5 mM) and within 300 s to the average glucose concentrations found in tear fluid (0.15 mM). We find unusual temperature-dependent kinetics, which derive from glucose mutarotation in solution. It is shown that alpha-d-glucose is the glucose anomer binding to the boronic acid derivative. Care must be taken in any glucose determination to ensure that the glucose mutarotation equilibrium has been established. We have demonstrated that the sensor is responsive to approximately 0.15 mM glucose concentrations in artificial tear fluid solution.

Lacrimal Duct Occlusion for the Treatment of Dry Eye

The most widespread treatment for dry eyes in clinical practice is an instillation containing artificial tear fluid. When an instillation does not ameliorate dry eye, we use punctal plugs. The insertion of punctal plugs is an eminent curative that positively improves the corneal and conjunctival epithelial disorders and the amount of lacrimal fluid accumulated in the conjunctival sac. We shall introduce the lacrimal duct occlusion utilizing atelocollagen solutions, which solves many of the issues induced by conventional methods.

Polyclonal human antibodies reduce bacterial attachment to soft contact lens and corneal cell surfaces

Bacterial keratitis due to Pseudomonas aeruginosa is a potentially serious complication of extended-wear contact lens use. Adhesion of P. aeruginosa to soft contact lens materials or corneal endothelial cells in the presence of pooled human immunoglobulins and/or neutrophils in artificial tear fluid was studied in vitro as a potential method to treat contact lens-associated infection. Soft hydrophilic contact lens materials equilibrated in sterile saline were soaked in artificial tear fluid for 18 h prior to use. P. aeruginosa IFO 3455 was added to groups of lenses or confluent cultured bovine corneal endothelial cells with varying amounts of human polyclonal immunoglobulin (IgG) and human blood neutrophils or serum albumin as a control. After 2 or 4 h incubation, adherent viable bacteria on lenses were quantified. Fluorescence microscopy was used to assess bacterial adherence to bovine corneal endothelial cells in the presence and absence of IgG and neutrophils. Various concentrations of albumin had no effect on adhesion. Human immunoglobulin solutions (25 mg/ml) reduced P. aeruginosa adhesion by nearly 1 log and 2 logs after 2 and 4 h incubations, respectively. Neutrophils in combination with 25 mg/ml IgG reduced bacterial adhesion approximately 1 log over reduction in adhesion by neutrophils alone. Diluted human IgG (10 mg/ml) did not significantly decrease bacterial adhesion after 2 or 4 h, but did reduce adhesion in combination with human neutrophils at both time points. Similar reductions in amounts of fluorescently labeled bacteria adhered to cultured monolayers of corneal endothelial cells under these conditions were qualitatively observed.

The role of different phospholipids on tear break-up time using a model eye

Purpose. The effect of different phospholipids in stabilising the tear film was investigated to determine if particular polar head groups gave greater stability than others. Methods. Purified phosphatidylcholine (PC), lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylserine (PS) and cardiolipin (CL) were used. These were applied to a model eye loaded with an artificial tear fluid and the tear breakup time (TBUT) was measured. Three variants of the artificial tear fluid were utilised: buffered saline alone; one with proteins and mucins; and one containing proteins, mucins and lipids. Results. TBUT was improved by the presence of phospholipids. In particular, the best performance was with PI applied to artificial tear fluid containing proteins mucins and lipids. Use of buffered saline as the artificial tear fluid gave very short break-up times. Conclusion. Increase in tear film stability by phospholipids is probably not due to the charge carried by the polar head group, but more likely due to the charge distribution, and the presence of hydroxyl groups in the head group also tends to increase stability, possibly through specific interactions with proteins and mucins in the subphase.

Surface protein profile of extended-wear silicon hydrogel lenses.

Protein deposition onto the surfaces of contact lenses during wear is problematic since it promotes bacterial adhesion 1and growth. 2 For instance, when Etafilcon A or Polymacon contact lenses are surface-coated with albumin, adhesion ofP. aeruginosaincreases.1 When Etafilcon A lenses are coated with an artificial tear fluid, the adhesion of some strains ofSerratia marcescensis enhanced, 2 and when lysozyme is adsorbed to these lenses, they exhibit greater adhesion ofStaphylococcus aureus. 3Lens deposits may also be involved in the production of giant papillary conjunctivitis.4Furthermore, over a long time period, protein deposition can result in reduced visual acuity.

Bacterial interactions with contact lenses; effects of lens material, lens wear and microbial physiology

Contact lens wear is a successful form of vision correction. However, adverse responses can occur during wear. Many of these adverse responses are produced as a consequence of bacterial colonization of the lens. The present study demonstrated that during asymptomatic contact lens wear lenses are colonized by low levels of bacteria with gram-positive bacteria, such as coagulase negative staphylococci, predominating. Gram-negative bacteria are frequently the causative agents of adverse responses during contact lens wear. Measuring the adhesion of different strains and/or species of bacteria to different contact lens materials demonstrated considerable differences. In particular, Pseudomonas aeruginosa strains Paer1 and 6294 and Aeromonas hydrophilia strain Ahyd003 adhered in larger numbers to the highly oxygen permeable contact lenses Balafilcon A compared to hydrogel lenses manufactured from either Etafilcon A or HEMA. Furthermore, after Balafilcon A lenses had been worn for 6 h during the day bacteria were able to adhere in greater numbers to the worn lenses compared to the unworn lenses with increases in adhesion ranging from 243% to 1393%. However, wearing Etafilcon A lenses usually resulted in a decrease in adhesion (22–48%). Bacteria were able to grow after adhesion to lenses soaked in artificial tear fluid and formed biofilms, visualized by scanning confocal microscopy. Chemostat grown bacterial cultures were utilized to enable control of bacterial growth conditions and bacteria were shown to adhere in the greatest numbers if grown under low temperature (25°C compared to 37°C). The changes in growth temperature was shown, using 2D gel electrophoresis, to change the experssion of cell-surface proteins and, using 1D gel electrophoresis, to change the expression of surface lipopolysaccharide of P. aeruginosa Paer1. Thus, these surface changes would have been likely to have mediated the increased adhesion to Etafilcon A contact lenses.

Fluid-Holding Time Parameter and Fluid Holding Capability of Polymeric Materials

The fluid holding capability of polymeric materials was semiquantitatively characterized with a new parameter called the fluid holding time (FHT). FHT measures the time it takes a continuous film to recede down the side of a Wilhelmy plate, which varies on different surfaces with the same degree of wettability. Both pure water and an artificial tear fluid (i.e., protein/salt solution) were used to systematically wet uncoated and plasma polymer coated polymeric plates. The FHT was calculated from Wilhelmy force loop plots in the event that a continuous water or aqueous film adhered to a polymeric surface. Such events are easily verified by consecutive wetting. Dynamic Wilhelmy force measurement is probably the most sensitive technique for monitoring wettability and surface configuration change, two important phenomenological factors that affect the FHT. The FHT measured by the Wilhelmy balance method can be effectively used to compare the liquid holding capabilities of different surfaces. The value of the ...

Synergic antistaphylococcal properties of lactoferrin and lysozyme.

Staphylococcus epidermidis colonises a wide range of implanted prosthetic devices, but rarely contact lenses -- despite a similarity in material composition. A conceivable explanation for this anomaly is the action of the tear defences, including the constitutive proteins lactoferrin and lysozyme. Therefore this study investigated the effect of lactoferrin, lysozyme and serum on the growth of S. epidermidis isolates in artificial tear fluid. Whether supplemented with serum alone or serum with either apolactoferrin or lysozyme, this medium induced a similar, strain-variable effect. However, simultaneous addition of these proteins induced a greater bactericidal or bacteristatic effect. Of those strains killed by the concerted action of apolactoferrin and lysozyme, the absence of serum led to a further increase in the bactericidal effect, whereas strains displaying bacteriostasis were unaffected by serum. Iron saturation of lactoferrin reversed the antimicrobial synergy of apolactoferrin and lysozyme. These results show synergy between lactoferrin and lysozyme which is dependent on the iron limitation of lactoferrin. As a bactericidal mechanism, this synergy is augmented by serum, but bacteriostasis remains unaffected by serum supplemention. Thus, the combination of lysozyme and lactoferrin may partly explain the low level of contact lens colonisation by S. epidermidis in vivo.

Corneal permeability of aldose reductase inhibitor, M79175. I. In vitro study using corneal permeability apparatus.

Transcorneal permeability of M79175, aldose reductase inhibitor (Eisai), and various factors influencing it were examined using excised rabbit cornea mounted on the methacrylate apparatus. High permeability was exhibited and this was independent of the concentration of M79175. Cornea hydrated by exposure to the eye drop solution of high concentration of benzalkonium chloride, which had an enhancing effect on the permeability of M79175. Corneal hydration was also caused by the solution of low pH. These findings will show the basic conditions for this eye drop solution to be an artificial tear fluid.