Low cytotoxicity, antibacterial property, and curcumin delivery performance of toughness-enhanced electrospun composite membranes based on poly(lactic acid) and MAX phase (Ti3AlC2).

MXenes, synthesized from their precursor MAX phases, have been extensively researched as additives to enhance the drug delivery performance of polymer matrices, whereas there is a limited number of previous reports on the use of MAX phases themselves for such applications. The use of MAX phases can exclude the complicated synthesis procedure and lessen resultant production and environmental costs required to convert MAX phases to MXenes. Herein, electrospun membranes of poly(lactic acid) (PLA) and a MAX phase (Ti3AlC2) have been fabricated for curcumin delivery. The composite membrane exhibits significantly higher toughness (8.82 MJ m-3) than the plasticized PLA membrane (0.63 MJ m-3) with low cytotoxicity, supporting proliferation of mouse fibroblast L929 cells. The curcumin-loaded composite membrane exhibits high water vapor transmission (∼7350 g m-2 day-1), porosity (∼85 %), water wettability, and antibacterial properties against E. coli and S. aureus. Seven-day curcumin release is enhanced from 45 % (PLA) to 67 % (composite) due to curcumin diffusion from the polymer fibers and MAX phase surface that contributes to overall increased curcumin adsorption and release sites. This work demonstrates the potential of the MAX phase to enhance both properties and curcumin delivery, promising for other eco-friendly systems for sustainable drug delivery applications.

Encapsulation of propolis extracts in aqueous formulations by using nanovesicles of lipid and poly(styrene-alt-maleic acid).

Bee propolis has been used in alternative medicine to treat various diseases. Due to its limited water solubility, it is often used in combination with alcohol solvents, causing skin irritation and immune response. To solve this, the new drug delivery system, based on the lipid nanodiscs of 1,2-dimyristoyl-sn-glycero-3-phosphochline (DMPC) and poly(styrene-alt-maleic acid) (PSMA), were created in an aqueous media. At the excess polymer concentrations, the PSMA/DMPC complexation produced the very fine nanoparticles (18 nm). With the increased molar ratio of styrene to maleic acid (St/MA) in the copolymer structure, the lipid nanodisc showed the improved encapsulation efficiency (EE%), comparing to their corresponding aqueous formulations. The maximum value had reached to around 20% when using the 2:1 PSMA precursor. Based on the cytotoxicity test, these nanoparticles were considered to be non-toxic over the low dose administration region (<78 µg/mL). Instead, they possessed the ability to promote the Vero cell growth. The new PSMA/DMPC nanovesicles could thus be used to improve aqueous solubility and therapeutic effects of poorly water-soluble drugs, thus extending their use in modern therapies.

New biomimetic approach for propolis encapsulation was developed with no use of organic solvent.Propolis antioxidants were recovered directly into water-soluble formats.The very fine lipid nanodiscs showed impressive shelf-life stability and tuneable drug-loading capacity.

In Situ Compatibilized Blends of PLA/PCL/CAB Melt-Blown Films with High Elongation: Investigation of Miscibility, Morphology, Crystallinity and Modelling.

Ternary-blended, melt-blown films of polylactide (PLA), polycaprolactone (PCL) and cellulose acetate butyrate (CAB) were prepared from preliminary miscibility data using a rapid screening method and optical ternary phase diagram (presented as clear, translucent, and opaque regions) as a guide for the composition selection. The compositions that provided optically clear regions were selected for melt blending. The ternary (PLA/PCL/CAB) blends were first melt-extruded and then melt-blown to form films and characterized for their tensile properties, tensile fractured-surface morphology, miscibility, crystallinity, molecular weight and chemical structure. The results showed that the tensile elongation at the break (%elongation) of the ternary-blended, melt-blown films (85/5/10, 75/10/15, 60/15/25 of PLA/PCL/CAB) was substantially higher (>350%) than pure PLA (ca. 20%). The range of compositions in which a significant increase in %elongation was observed at 55−85% w/w PLA, 5−20% w/w PCL and 10−25% w/w CAB. Films with high %elongation all showed good interfacial interactions between the dispersed phase (PCL and CAB) and matrix (PLA) in FE-SEM and showed improvements in miscibility (higher intermolecular interaction and mixing) and a decrease in the glass transition temperature, when compared to the low %elongation films. The decrease in Mw and Mn and the formation of the new NMR peaks (1H NMR at 3.68−3.73 ppm and 13C NMR at 58.54 ppm) were observed in only the high %elongation films. These are expected to be in situ compatibilizers that are generated during the melt processing, mostly by chain scission. In addition, mathematical modelling was used to study the optimal ratio and cost-effectiveness of blends with optimised mechanical properties. These ternary-blended, melt-blown films have the potential for use in both packaging and medical devices with excellent mechanical performance as well as inherent economic and environmental capabilities.

A study of the permeation and water-structuring behavioural properties of PEG modified hydrated silk fibroin membranes.

The potential of naturally occurring substances as a source of biomedical materials is well-recognised and is being increasingly exploited. Silk fibroin membranes derived fromBombyx morisilk cocoons exemplify this, for example as substrata for the growth of ocular cells with the aim of generating biomaterial-cell constructs for tissue engineering. This study investigated the transport properties of selected silk fibroin membranes under conditions that allowed equilibrium hydration of the membranes to be maintained. The behaviour of natural fibroin membranes was compared with fibroin membranes that have been chemically modified with poly(ethylene glycol). The permeation of the smaller hydrated sodium ion was higher than that of the hydrated calcium ion for all three ethanol treated membranes investigated. The PEG and HRP-modified C membrane, which had the highest water content at 59.6 ± 1.5% exhibited the highest permeation of the three membranes at 95.7 ± 2.8 × 10-8cm2s-1compared with 17.9 ± 0.9 × 10-8cm2s-1and 8.7 ± 1.7 × 10-8cm2s-1for membranes A and B respectively for the NaCl permeant. Poly(ethylene glycol) was used to increase permeability while exploiting the crosslinking capabilities of horseradish peroxidase to increase the compressive strength of the membrane. Importantly, we have established that the permeation behaviour of water-soluble permeants with hydrated radii in the sub-nanometer range is analogous to that of conventional hydrogel polymers.

The influence of structure and morphology on ion permeation in commercial silicone hydrogel contact lenses.

The importance of the microstzructure of silicone hydrogels is widely appreciated but is poorly understood and minimally investigated. To ensure comfort and eye health, these materials must simultaneously exhibit both high oxygen and high water permeability. In contrast with most conventional hydrogels, the water content and water structuring within silicone hydrogels cannot be solely used to predict permeability. The materials achieve these opposing requirements based on a composite of nanoscale domains of oxygen-permeable (silicone) and water-permeable hydrophilic components. This study correlated characteristic ion permeation coefficients of a selection of commercially available silicone hydrogel contact lenses with their morphological structure and chemical composition. Differential scanning calorimetry measured the water structuring properties through subdivision of the freezing water component into polymer-associated water (loosely bound to the polymer matrix) and ice-like water (unimpeded with a melting point close to that of pure water). Small-angle x-ray scattering, and environmental scanning electron microscopy techniques were used to investigate the structural morphology of the materials over a range of length scales. Significant, and previously unrecognized, differences in morphology between individual materials at nanometer length scales were determined; this will aid the design and performance of the next generation of ocular biomaterials, capable of maintaining ocular homeostasis.

A comparison of SMA (styrene maleic acid) and DIBMA (di-isobutylene maleic acid) for membrane protein purification.

The use of styrene maleic acid co-polymer (SMA) for membrane protein extraction and purification has grown in recent years. SMA inserts in the membrane and assembles into small discs of bilayer encircled by polymer, termed SMA lipid particles (SMALPs). This allows purification of membrane proteins whilst maintaining their lipid bilayer environment. SMALPs offer several improvements over conventional detergent approaches, however there are limitations, most notably a sensitivity to low pH and divalent cations. Recently it was shown that the aliphatic diisobutylene-maleic acid (DIBMA) copolymer, was also able to directly solubilise membranes forming DIBMALPs (DIBMA lipid particles), and that this polymer overcame some of the limitations of SMA. In this study the ability of DIBMA to solubilise and purify functional membrane proteins has been compared to SMA. It was found that DIBMA is able to solubilise several different membrane proteins from different expression systems, however for some proteins it gives a lower yield and lower degree of purity than SMA. DIBMA extracted G protein-coupled receptors retain ligand- and G protein-binding. DIBMALPS are larger than SMALPs and display a decreased sensitivity to magnesium. However the stability of DIBMALPs appears to be lower than SMALPs. The lower purity and lower stability are likely linked to the larger size of the DIBMALP particle. However, this also offers a potentially less rigid lipid environment which may be more amenable to protein dynamics. Therefore the optimal choice of polymer will depend on which features of a protein are to be investigated.

Clinical and biochemical analysis of the ageing tear film.

BACKGROUND: Tear film stability is important for healthy visual function, and yet little is known of the ageing mechanisms. The aim of this study was to investigate parallels between biochemical changes and clinical physical parameters, which occur in the tear film of two subject populations differing in age by over 30 years. METHODS: Two distinct age groups were chosen: 11 'younger' (23.7±2.1 years) and 19 'older' (63.0±4.0 years) subjects. A series of clinical tests were performed to access tear volume, tear film stability and general ocular health. Tear protein analyses from extracted Schirmer strips were conducted with the Agilent 2100 Bioanalyzer. RESULTS: Clinical investigations highlighted significant differences between the age groups. For example: McMonnies scores (p=0.009) and bulbar redness (p=0.038) were higher for the older group, whereas tear meniscus height was larger (p=0.018) in the younger group. Similarly, relative plasma-derived albumin levels were higher (17.1%±12.4%) in the tears of the older, compared with the younger (5.0%±9.6%) group. A protein peak at ∼23 kDa was observed in 53% of the older group samples but in only 36% of the samples of the younger subjects (p=0.122). CONCLUSIONS: Distinct differences in tear film composition between the two age groups were observed. Parallels in terms of clinical symptoms which reflected a biochemical response (and vice versa) were found, but specific correlations between clinical measurements and biomarkers for individual subjects were not observed.

High modulus hydrogels for ophthalmic and related biomedical applications.

This paper presents three families of semi-interpenetrating polymer network (SIPN) hydrogels based on an ester-based polyurethane (EBPU) and hydrophilic monomers: N,N-dimethylacrylamide (NNDMA), N-vinyl pyrrolidone (NVP) and acryloylmorpholine (AMO) as potential materials for keratoprosthesis, orthokeratology and mini-scleral lens application. Hydrogels sheets were synthesized via free-radical polymerization with methods developed in-house. SIPN hydrogels were characterized for their equilibrium water content, mechanical and surface properties. Three families of optically clear SIPN-based hydrogels have been synthesized in the presence of water with >10% of composition attributable to EBPU. Water contents of SIPN materials ranged from 30% to 70%. SIPNs with ≤15% EBPU of total composition showed little influence to mechanical properties, whereas >15% EBPU contributed significantly to an increase in material stiffness. In the hydrated state, SIPNs with ≤15% EBPU of total composition show little difference in polar component (γp ) of surface free energy, whereas for >15% EBPU there is a decrease in γp . The EBPU SIPN hydrogels display complementary material properties for keratoprosthesis, orthokeratology, and mini-scleral applications. © 2018 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1645-1653, 2019.

Hydrophobic and hydrophilic effects on water structuring and adhesion in denture adhesives.

Denture adhesives are designed to be moisture-sensitive through the inclusion of a blend of polymer salts with varying degrees of water-sensitivity. This enables the adhesive to mix with saliva in vivo and activate its high tack, through the formation of a mucilaginous layer. We report for the first time, the use of differential scanning calorimetry to study a series of hydrophobic and hydrophilic polymeric systems in order to correlate water-structuring behavior with adhesion strength. Adhesive bonding of the more hydrophobic variants was higher than that of a commercial-based control and a more hydrophilic polymer system in both lap shear and tensile configurations. Water-binding data suggested that increasing the hydrophobicity of the maleic acid copolymer substituents led to decreased levels of freezing water. In comparison, increasing the hydrophilic nature of the polymer backbone gave higher levels of freezing water within the hydrated samples. The results of this study emphasize the importance of varying the levels of hydrophobic and hydrophilic components within denture adhesive formulations, alongside the types of water present within the adhesive systems. This phenomenon has shown the potential to fine-tune the adhesive properties and failure mode against poly(methyl methacrylate), surfaces. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1355-1362, 2018.

Tunable denture adhesives using biomimetic principles for enhanced tissue adhesion in moist environments.

Nature provides many interesting examples of adhesive strategies. Of particular note, the protein glue secreted by marine mussels delivers high adhesion in wet and dynamic environments owing to existence of catechol moieties. As such, this study focuses on denture fixatives, where a non-zinc-containing commercial-based formulation has been judiciously modified by a biomimetic catechol-inspired polymer, poly(3,4-dihydroxystyrene/styrene-alt-maleic acid) in a quest to modulate adhesive performance. In vitro studies, in a lap-shear configuration, revealed that the catechol-modified components were able to enhance adhesion to both the denture base and hydrated, functional oral tissue mimic, with the resulting mode of failure prominently being adhesive rather than cohesive. These characteristics are desirable in prosthodontic fixative applications, for which temporary adhesion must be maintained, with ultimately an adhesive failure from the mucosal tissue surface preferred. These insights provide an experimental platform in the design of future biomimetic adhesive systems. STATEMENT OF SIGNIFICANCE: Mussel adhesive proteins have proven to be promising biomimetic adhesive candidates for soft tissues and here for the first time we have adapted marine adhesive technology into a denture fixative application. Importantly, we have incorporated a soft tissue mimic in our in vitro adhesion technique that more closely resembles the oral mucosa than previously studied substrates. The novel biomimetic-modified adhesives showed the ability to score the highest adhesive bonding out of all the formulations included in this study, across all moisture levels. This paper will be of major interest to the Acta Biomaterialia readership since the study has illustrated the potential of biomimetic principles in the design of effective prosthodontic tissue adhesives in a series of purpose-designed in vitro experiments in the context of the challenging features of the oral environment.

Mechanical properties of contact lenses: The contribution of measurement techniques and clinical feedback to 50 years of materials development.

PURPOSE: This review summarises the way in which mechanical property measurements combined with clinical perception have influenced the last half century of materials evolution in contact lens development. METHODS: Literature concerning the use of in-vitro testing in assessment of the mechanical behaviour of contact lenses, and the mutual deformation of the lens material and ocular tissue was examined. Tensile measurements of historic and available hydrogel lenses have been collected, in addition to manufacturer-generated figures for the moduli of commercial silicone hydrogel lenses. RESULTS: The three conventional modes of mechanical property testing; compression, tension and shear each represent different perspective in understanding the mutual interaction of the cornea and the contact lens. Tensile testing provides a measure of modulus, together with tensile strength and elongation to break, which all relate to handling and durability. Studies under compression also measure modulus and in particular indicate elastic response to eyelid load. Studies under shear conditions enable dynamic mechanical behaviour of the material to be assessed and the elastic and viscous components of modulus to be determined. These different methods of measurement have contributed to the interpretation of lens behaviour in the ocular environment. An amalgamated frequency distribution of tensile moduli for historic and currently available contact lens materials reveals the modal range to be 0.3-0.6MPa. CONCLUSION: Mechanical property measurements of lens materials have enabled calibration of an important aspect of their ocular interaction. This together with clinical feedback has influenced development of new lens materials and assisted clinical rationalisation of in-eye behaviour of different lenses.

Structural design of contact lens-based drug delivery systems; in vitro and in vivo studies of ocular triggering mechanisms.

This study identifies and investigates the potential use of in-eye trigger mechanisms to supplement the widely available information on release of ophthalmic drugs from contact lenses under passive release conditions. Ophthalmic dyes and surrogates have been successfully employed to investigate how these factors can be drawn together to make a successful system. The storage of a drug-containing lens in a pH lower than that of the ocular environment can be used to establish an equilibrium that favours retention of the drug in the lens prior to ocular insertion. Although release under passive conditions does not result in complete dye elution, the use of mechanical agitation techniques which mimic the eyelid blink action in conjunction with ocular tear chemistry promotes further release. In this way differentiation between passive and triggered in vitro release characteristics can be established. Investigation of the role of individual tear proteins revealed significant differences in their ability to alter the equilibrium between matrix-held and eluate-held dye or drug. These individual experiments were then investigated in vivo using ophthalmic dyes. Complete elution was found to be achievable in-eye; this demonstrated the importance of that fraction of the drug retained under passive conditions and the triggering effect of in-eye conditions on the release process. Understanding both the structure-property relationship between drug and material and in-eye trigger mechanisms, using ophthalmic dyes as a surrogate, provides the basis of knowledge necessary to design ocular drug delivery vehicles for in-eye release in a controllable manner.

The design of contact lens based ocular drug delivery systems for single-day use: Part (I) Structural factors, surrogate ophthalmic dyes and passive diffusion studies.

The poor retention and efficacy of instilled drops as a means of delivering drugs to the ophthalmic environment is well-recognised. The potential value of contact lenses as a means of ophthalmic drug delivery, and consequent improvement of pre-corneal retention is one obvious route to the development of a more effective ocular delivery system. Furthermore, the increasing availability and clinical use of daily disposable contact lenses provides the platform for the development of viable single-day use drug delivery devices based on existing materials and lenses. In order to provide a basis for the effective design of such devices, a systematic understanding of the factors affecting the interaction of individual drugs with the lens matrix is required. Because a large number of potential structural variables are involved, it is necessary to achieve some rationalisation of the parameters and physicochemical properties (such as molecular weight, charge, partition coefficients) that influence drug interactions. Ophthalmic dyes and structurally related compounds based on the same core structure were used to investigate these various factors and the way in which they can be used in concert to design effective release systems for structurally different drugs. Initial studies of passive diffusional release form a necessary precursor to the investigation of the features of the ocular environment that over-ride this simple behaviour. Commercially available contact lenses of differing structural classifications were used to study factors affecting the uptake of the surrogate actives and their release under 'passive' conditions. The interaction between active and lens material shows considerable and complex structure dependence, which is not simply related to equilibrium water content. The structure of the polymer matrix itself was found to have the dominant controlling influence on active uptake; hydrophobic interaction with the ophthalmic dye playing a major role.

Evidence of lipid degradation during overnight contact lens wear: gas chromatography mass spectrometry as the diagnostic tool.

PURPOSE: We investigated structural differences in the fatty acid profiles of lipids extracted from ex vivo contact lenses by using gas chromatography mass spectrometry (GCMS). Two lens materials (balafilcon A or lotrafilcon A) were worn on a daily or continuous wear schedule for 30 and 7 days. METHODS: Lipids from subject-worn lenses were extracted using 1:1 chloroform: methanol and transmethylated using 5% sulfuric acid in methanol. Fatty acid methyl esters (FAMEs) were collected using hexane and water, and analyzed by GCMS (Varian 3800 GC, Saturn 2000 MS). RESULTS: The gas chromatograms of lens extracts that were worn on a continuous wear schedule showed two predominant peaks, C16:0 and C18:0, both of which are saturated fatty acids. This was the case for balafilcon A and lotrafilcon A lenses. However, the gas chromatograms of lens extracts that were worn on a daily wear schedule showed saturated (C16:0, C18:0) and unsaturated (C16:1 and C18:1) fatty acids. CONCLUSIONS: Unsaturated fatty acids are degraded during sleep in contact lenses. Degradation occurred independently of lens material or subject-to-subject variability in lipid deposition. The consequences of lipid degradation are the production of oxidative products, which may be linked to contact lens discomfort.

Structural effects in photopolymerized sodium AMPS hydrogels crosslinked with poly(ethylene glycol) diacrylate for use as burn dressings.

Synthetic hydrogel polymers were prepared by free radical photopolymerization in aqueous solution of the sodium salt of 2-acrylamido-2-methylpropane sulfonic acid (Na-AMPS). Poly(ethylene glycol) diacrylate (PEGDA) and 4,4'-azo-bis(4-cyanopentanoic acid) were used as the crosslinker and UV-photoinitiator, respectively. The effects of varying the Na-AMPS monomer concentration within the range of 30-50% w/v and the crosslinker concentration within the range of 0.1-1.0% mol (relative to monomer) were studied in terms of their influence on water absorption properties. The hydrogel sheets exhibited extremely high swelling capacities in aqueous media which were dependent on monomer concentration, crosslink density, and the ionic strength and composition of the immersion medium. The effects of varying the number-average molecular weight of the PEGDA crosslinker from [Formula: see text] = 250 to 700 were also investigated. Interestingly, it was found that increasing the molecular weight and therefore the crosslink length at constant crosslink density decreased both the rate of water absorption and the equilibrium water content. Cytotoxicity testing by the direct contact method with mouse fibroblast L929 cells indicated that the synthesized hydrogels were nontoxic. On the basis of these results, it is considered that photopolymerized Na-AMPS hydrogels crosslinked with PEGDA show considerable potential for biomedical use as dressings for partial thickness burns. This paper describes some structural effects which are relevant to their design as biomaterials for this particular application.

A decade of silicone hydrogel development: surface properties, mechanical properties, and ocular compatibility.

Since the initial launch of silicone hydrogel lenses, there has been a considerable broadening in the range of available commercial material properties. The very mobile silicon-oxygen bonds convey distinctive surface and mechanical properties on silicone hydrogels, in which advantages of enhanced oxygen permeability, reduced protein deposition, and modest frictional interaction are balanced by increased lipid and elastic response. There are now some 15 silicone hydrogel material variants available to practitioners; arguably, the changes that have taken place have been strongly influenced by feedback based on clinical experience. Water content is one of the most influential properties, and the decade has seen a progressive rise from lotrafilcon-A (24%) to efrofilcon-A (74%). Moduli have decreased over the same period from 1.4 to 0.3 MPa, but not solely as a result of changes in water content. Surface properties do not correlate directly with water content, and ingenious approaches have been used to achieve desirable improvements (e.g., greater lubricity and lower contact angle hysteresis). This is demonstrated by comparing the hysteresis value of the earliest (lotrafilcon-A, >40°) and most recent (delefilcon-A, <10°) coated silicone hydrogels. Although wettability is important, it is not of itself a good predictor of ocular response because this involves a much wider range of physicochemical and biochemical factors. The interference of the lens with ocular dynamics is complex leading separately to tissue-material interactions involving anterior and posterior lens surfaces. The biochemical consequences of these interactions may hold the key to a greater understanding of ocular incompatibility and end of day discomfort.

Function of lipids − their fate in contact lens wear: an interpretive review.

Lipids play a vital role in the body at many interfaces. Examples include the lubrication of articulating joints by synovial fluid, the coating of the lung by pulmonary surfactant and the functions of the tear film in the protection of the anterior eye. The role of the lipids is similar at each site - acting as boundary lubricants and reducing surface and interfacial tension. This review focuses on how and why contact lens wear can disrupt the normal function of lipids within the tear film and explains how the otherwise advantageous presence and function of tear lipids can become disadvantageous, causing problems for the wearer. Because the contact lens is some ten times thicker than the tear film, lipids deposited on the anterior surface become immobilised, reducing lipid turnover and thus leading to prolonged exposure to oxygen and light with consequent generation of degradation products. These degraded lipids reduce lens wettability and have additionally been linked to problems of contact lens discomfort and intolerance. Lipid problems are influenced by the thickness of the lens, the material, surface modification, mode of wear and ultimately the subject. The most influential of these variables is frequently the subject.

Tear analysis and lens-tear interactions: part II. Ocular lipids-nature and fate of meibomian gland phospholipids.

PURPOSE: Published data indicate that the polar lipid content of human meibomian gland secretions (MGS) could be anything between 0.5% and 13% of the total lipid. The tear film phospholipid composition has not been studied in great detail and it has been understood that the relative proportions of lipids in MGS would be maintained in the tear film. The purpose of this work was to determine the concentration of phospholipids in the human tear film. METHODS: Liquid chromatography mass spectrometry (LCMS) and thin layer chromatography (TLC) were used to determine the concentration of phospholipid in the tear film. Additionally, an Amplex Red phosphatidylcholine-specific phospholipase C (PLC) assay kit was used for determination of the activity of PLC in the tear film. RESULTS: Phospholipids were not detected in any of the tested human tear samples with the low limit of detection being 1.3 µg/mL for TLC and 4 µg/mL for liquid chromatography mass spectrometry. TLC indicated that diacylglycerol (DAG) may be present in the tear film. PLC was in the tear film with an activity determined at approximately 15 mU/mL, equivalent to the removal of head groups from phosphatidylcholine at a rate of approximately 15 µM/min. CONCLUSIONS: This work shows that phospholipid was not detected in any of the tested human tear samples (above the lower limits of detection as described) and suggests the presence of DAG in the tear film. DAG is known to be at low concentrations in MGS. These observations indicate that PLC may play a role in modulating the tear film phospholipid concentration.

Tear analysis and lens-tear interactions. Part I. Protein fingerprinting with microfluidic technology.

The purpose of this work is to establish the application of a fully automated microfluidic chip based protein separation assay in tear analysis. It is rapid, requires small sample volumes and is vastly superior to, and more convenient than, comparable conventional gel electrophoresis assays. The protein sizing chip technology was applied to three specific fields of analysis. Firstly tear samples were collected regularly from subjects establishing the baseline effects of tear stimulation, tear state and patient health. Secondly tear samples were taken from lens wearing eyes and thirdly the use of microfluidic technology was assessed as a means to investigate a novel area of tear analysis, which we have termed the 'tear envelope'. Utilising the Agilent 2100 Bioanalyzer in combination with the Protein 200 Plus LabChip kit, these studies investigated tear proteins in the range of 14-200 kDa. Particular attention was paid to the relative concentrations of lysozyme, tear lipocalin, secretory IgA (sIgA), IgG and lactoferrin, together with the overall tear electropherogram 'fingerprint'. Furthermore, whilst lens-tear interaction studies are generally thought of as an investigation into the effects of tears components on the contact lens material, i.e. deposition studies, this report addresses the reverse phenomenon--the effect of the lens, and particularly the newly inserted lens, on the tear fluid composition and dynamics. The use of microfluidic technology provides a significant advance in tear studies and should prove invaluable in tear diagnostics and contact lens performance analysis.