M. Elisabete Oliveira

Contact Lens Hydrophobicity and Roughness Effects on Bacterial Adhesion

Purpose. This study was designed to assess whether hydrogel contact lens (CL) surface hydrophobicity and roughness affect Staphylococcus epidermidis adhesion. Methods. Bacterial adhesion experiments were performed on two unworn silicone hydrogel and three unworn conventional hydrogel CLs using the S.epidermidis strain CECT 4184. Microbial colonization was assessed by conducting counts expressed as colony-forming units. CL hydrophobicity was determined through water contact angle measurements and the roughness parameters such as mean surface roughness (Ra), kurtosis (Rku), and skewness (Rsk) were determined through atomic force microscopy in Tapping Mode. Results. The conventional CLs showed similar water contact angles (p > 0.05) and were classified as hydrophilic. The silicone hydrogel CLs yielded hydrophobic contact angles with no significant differences between them (p > 0.05). The lenses with the highest (nelfilcon A and ocufilcon B) or lowest (comfilcon A and omafilcon A) Ra values displayed a lesser or greater extent of spikiness of their surfaces, respectively. All lenses showed a predominance of peaks (Rsk > 0) over troughs. S. epidermidis adhered more to the hydrophobic CLs (p < 0.05). Omafilcon A and comfilcon A, which showed the lowest Ra values among the hydrophilic and hydrophobic lenses, respectively, returned the lowest bacterial adhesion scores (p < 0.05). Conclusions. Our results suggest that more hydrophobic CLs are more prone to S. epidermidis adhesion. Although the Ra appears to be related to S. epidermidis adhesion, the influence of Rku and Rsk on this variable remains unclear.

Soft Contact Lens Surface Profile by Atomic Force Microscopy

Purpose. This study was designed to qualitatively and quantitatively characterize the surface morphology of four unworn conventional hydrogel contact lenses (omafilcon, hioxifilcon, nelfilcon A, and ocufilcon B) and two silicone-hydrogel contact lenses (senofilcon A and comfilcon A) without surface treatment. Methods. Surface roughness was assessed using atomic force microscopy in Tapping ModeTM to determine the representative roughness parameters mean surface roughness (Ra), mean square roughness (Rms), kurtosis (Rku), and skewness (Rsk). To date, these last two parameters have not been used to characterize contact lens surfaces. Rku provides information on the distribution of spikes above and below the mean line, and Rsk provides information about the asymmetric roughness of surfaces. The surface topography of the lenses was also mapped in detail. Results. In all the lenses, higher Ra and Rms values were obtained when larger surface areas were examined. The daily replacement contact lenses (nelfilcon A and ocufilcon B) showed the highest Ra and Rms values but according to their Rku scores, their surface profiles were less spiky than the remaining lenses. On the contrary, the lowest Ra and Rms values were recorded for comfilcon A and omafilcon A, which also exhibited the spikiest surface profiles. All the materials except the hioxifilcon showed a predominance of peaks (Rsk >0) over troughs. Conclusions. The shape parameters Rku and Rsk are useful for characterizing contact lens surfaces, because they provide different yet complementary information to that offered by Ra and Rms. Precise knowledge of the shape profile of a contact lens surface will give an idea of its susceptibility to deposit formation or colonization by microorganisms.

Bacterial adhesion to worn silicone hydrogel contact lenses

Purpose. The aim of this study was to, firstly, investigate whether silicone-hydrogel contact lenses (CL) are more or less susceptible to bacterial adhesion than conventional ones and, secondly, assess the influence of lens wear in the extent of bacterial adhesion. Four silicone-hydrogel CL (galyfilcon A, balafilcon A, lotrafilcon A, and lotrafilcon B) and one conventional hydrogel (etafilcon A) CL were tested. Methods. Bacterial adhesion experiments were performed on unworn and worn CL using the strain Staphylococcus epidermidis 9142. Worn lenses were obtained from a group of 31 subjects fitted with a silicone-hydrogel CL in one eye and a conventional hydrogel CL as contralateral pair. These lenses were used on a daily basis in combination with a multipurpose lens care solution. Adhesion assays were carried out in a parallel plate flow chamber, followed by image analysis. Hydrophobicity, roughness, and topography of the lenses surfaces were assessed through contact angle measurements and atomic force microscopy. Results. Unworn conventional and silicone-hydrogel CL were equally susceptible to bacterial adhesion of S. epidermidis. Conversely, worn conventional hydrogel (etafilcon A) were more prone to bacterial adhesion than worn silicone-hydrogel materials, which exhibited similar adhesion extents among them. The results also showed that the lens surface properties such as hydrophobicity, roughness, and surface topography changed during wear. The alteration of surface hydrophobicity of silicone and conventional hydrogel CL during wear had a great impact on lens bacterial adhesion susceptibility. Accordingly, balafilcon A becomes significantly less hydrophobic and less prone to bacterial adhesion after lens wear, whereas etafilcon A becomes more hydrophobic and also more susceptible to bacterial adhesion (p < 0.05). Conclusions. Worn silicone-hydrogel galyfilcon A, balafilcon A, lotrafilcon A, and lotrafilcon B are equally prone to microbial adhesion of S. epidermidis and generally less susceptible than the conventional hydrogel.

Aggregation behavior of aqueous dioctadecyldimethylammonium bromide/monoolein mixtures: A multitechnique investigation on the influence of composition and temperature

A recently described non-viral gene delivery system [dioctadecyldimethylammonium bromide (DODAB)/monoolein (MO)] has been studied in detail to improve knowledge on the interactions between lamellar (DODAB) and non-lamellar-forming (MO) lipids, as a means to enhance their final cell transfection efficiency. Indeed, the morphology, fluidity, and size of these cationic surfactant/neutral lipid mixtures play an important role in the ability of these systems to complex nucleic acids. The different techniques used in this work, namely dynamic light scattering (DLS), fluorescence spectroscopy, differential scanning calorimetry (DSC), cryogenic transmission electron microscopy (cryo-TEM), light microscopy (LM), and surface pressure-area isotherms, allowed fully characterization of the phase behavior and aggregate morphology of DODAB/MO mixtures at different molar ratios. Overall, the results indicate that the final morphology of DODAB/MO aggregates depends on the balance between the tendency of DODAB to form zero-curvature bilayer structures and the propensity of MO to form non-bilayer structures with negative curvature. These results also show that in the MO-rich region, an increase in temperature has a similar effect on aggregate morphology as an increase in MO concentration.

DODAB:monoolein liposomes containing Candida albicans cell wall surface proteins: A novel adjuvant and delivery system

We describe the preparation and characterization of DODAB:MO-based liposomes and demonstrate their adjuvant potential and use in antigen delivery. Liposomes loaded with Candida albicans proteins assembled as stable negatively charged spherical nanoparticles with a mean size of 280 nm. High adsorption efficiency (91.0 ± 9.0%) is attained with high lipid concentrations. The nanoparticles were non-toxic, avidly taken up by macrophage cells and accumulated in membrane rich regions with an internalization time of 20 min. Immunized mice displayed strong humoral and cell-mediated immune responses, producing antibodies (IgGs) against specific cell wall proteins, Cht3p and Xog1p. DODAB:MO-based liposomes loaded with C. albicans proteins have an excellent immunogenic potential and can be explored for the development of an immunoprotective strategy against Candida infections.

Tunable pDNA/DODAB:MO lipoplexes : the effect of incubation temperature on pDNA/DODAB:MO lipoplexes structure and transfection efficiency

Dioctadecyldimethylammonium bromide (DODAB):1-monooleoyl-rac-glycerol (MO) cationic liposomes were reported as a promising alternative to common transfection agents, showing superior effectiveness on the transfection of the 293T mammalian cell line with pSV-β-gal plasmid DNA. The study of DODAB:MO aggregates in the absence of DNA has indicated that their morphology depends on the balance between DODABs tendency to form bilayer structures and MOs propensity to form inverted non-lamellar structures. Other parameters, such as the temperature have proved to be crucial in the definition of the morphology of the developed nanocarrier. Therefore, in this work, a step forward to the current gene carrier system will be given by studying the effect of the tunable parameters (incubation temperature and MO content) on the structure of pDNA:DODAB:MO lipoplexes. More importantly, the implications that these tunable parameters could have in terms of lipoplex transfection efficiency will be investigated. Dynamic light scattering (DLS), zeta (ζ) potential, cryo-transmission electron microscopy (cryo-TEM) and ethidium bromide (EtBr) exclusion were used to assess the formation, structure and destabilization of pDNA:DODAB:MO lipoplexes at DODAB molar fractions of (1:1) and above equimolarity (2:1, 4:1) prepared at incubation temperatures from 25 to 50°C. Experimental results indicate that pDNA:DODAB:MOs structure is sensitive to the lipoplex incubation temperature, resulting in particles of distinct size, superficial charge and structure. These variations are also visible on the complexation dynamics of pDNA, and subsequent release upon incubation with the model proteoglycan heparin (HEP), at 25 and 50°C. Increase in temperature leads to re-organization of DODAB and MO molecules within the liposomal formulation, causing a positive charge re-localization in the lipoplex surface, which not only alters its structure but also its transfection efficiency. Altogether, these results confirm that in the DODAB:MO carriers, an increase in the incubation temperature has a similar effect on aggregate morphology as the observed with an increase in MO content. This conclusion is extended to the pDNA:DODAB:MO lipoplexes morphology and subsequent transfection efficiency defining new strategies in lipoplexes preparation that could be used to modulate the properties of other lipid formulations for nonviral gene delivery applications.

The effect of octylglucoside and sodium cholate in Staphylococcus epidermidis and Pseudomonas aeruginosa adhesion to soft contact lenses

Purpose. In this study, the effect of the natural surfactants octylglucoside and sodium cholate in inhibiting Staphylococcus epidermidis and Pseudomonas aeruginosa adhesion to conventional and silicone-hydrogel contact lenses (CL) was assessed. Hydrophobicity was also evaluated to conditioned and nonconditioned CL. Methods. The inhibiting effect of the tested surfactants was determined through “in vitro” adhesion studies to conditioned and nonconditioned CL followed by image acquisition and cell enumeration. Hydrophobicity was evaluated through contact angle measurements using the advancing type technique on air. Results. Sodium cholate exhibits a very low capability to inhibit microbial adhesion. Conversely, octylglucoside effectively inhibited microbial adhesion in both types of lenses. This surfactant exhibited an even greater performance than a multipurpose lens care solution used as control. Octylglucoside was the only tested surfactant able to lower the hydrophobicity of all CL, which can explain its high performance. Conclusions. The results obtained in this study point out the potential of octylglucoside as a conditioning agent to prevent microbial colonization.

Comparison of the tear film clinical parameters at two different times of the day

Purpose:  The aim of the present study was to analyse the quality and quantity of the tear film in a young, healthy and non‐contact lens‐wearing population with measurements taken in the morning and in the afternoon to establish if changes exist in these parameters.

Comparison of IgA, TNF-α and surface tension of the tear film in two different times of the day

PURPOSE The main goal was to study the biochemical composition of the tear film in two different times of the day. METHODS Tear samples were collected from 10 individuals, non contact lenses wearers, from the university population without pathologies. To assess daily variations in the tear film, samples were collected twice in the day, one early in the morning and another in the evening using capillary tubes. Tear protein profile was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), tumor necrosis factor α (TNF-α) and immunoglobulin A (IgA) were quantified by Enzyme-Linked ImmunoSorbent Assay (ELISA). Tear film stability was obtained through measurements of Non-Invasive Break Up Time and tear surface tension was measured by obtaining the Langmuir isotherms. RESULTS The stability of the tear film was higher in the morning than in the afternoon; corresponding to a higher value of surface tension in the afternoon. Protein electrophoresis tear profile is variable during the day as IgA concentration decreased from morning to afternoon (p<0.05). TNF-α concentration also decreased, but there were not significant statistical differences (p=0.089). CONCLUSION We concluded that there are daily variations in the composition and properties of the tear film, indicating that changes occur without being caused by contact lenses wear or by the presence of ocular and systemic pathologies. Presence of TNF-α is not a direct indication of inflammatory pathology, since this cytokine was identified in relatively high amounts in subjects without inflammatory pathology.

White light interferometry to characterize the hydrogel contact lens surface

Purpose:  The aim of this study was to characterize, qualitatively and quantitatively, the surface morphology of four unworn conventional hydrogel contact lenses (Omafilcon, Hioxifilcon‐based, Nefilcon A and Ocufilcon B) by White Light Optical Profiling (WLOP). WLOP is an ideal technique for sampling larger areas as well as for higher measurement speed compared with other topography techniques used in contact lens studies.