Holly Lorentz

The impact of lipid on contact angle wettability.

Purpose. To analyze the effect of in vitro lipid doping on conventional hydrogel (CH) and silicone hydrogel (SH) lens wettability, assessed by sessile drop contact angle (CA) measurement. Methods. Nine contact lens materials, five SHs and four CH, were incubated with two different lipid tear solutions (LTS) containing cholesterol, cholesteryl oleate, oleic acid, oleic acid methyl ester, and triolein. The first LTS was a “low” concentration solution, which was close to human values, and the second was a “high” concentration. Lenses were soaked in the two LTS types for 2 or 5 days and compared with lenses soaked in phosphate buffered saline (PBS) only. After soaking, advancing CAs were measured on a customized computerized device using a sessile drop method. Results. Compared with PBS, CAs for untreated SHs were unaffected by soaking in the LTS, with typical CA values of >95° (p > 0.05). The surface-treated SH materials exhibited markedly reduced CAs after lipid exposure, with the high concentration LTS reducing the CA to <5° (p < 0.01). The CH materials all exhibited lower CAs after soaking, with values typically decreasing to 35°, which was significantly lower than that seen with PBS (p < 0.01). Conclusion. Exposure to lipid may improve the wettability of certain SH and CH materials, particularly those SH materials that are surface treated. This may help to explain why certain SH materials appear to improve in comfort for some patients during the first few hours or days of wear.

Rewetting Drops Containing Surface Active Agents Improve the Clinical Performance of Silicone Hydrogel Contact Lenses

Purpose. The purpose of this study was to investigate the impact of using a rewetting drop (RWD) containing surface active agents (OPTI-FREE RepleniSH; Alcon, Fort Worth, TX) on the clinical performance and protein deposition when using a continuous-wear (CW) silicone hydrogel (SH) contact lens. Methods. Subjects wore lotrafilcon A SH lenses on a 30-day CW basis for two consecutive 1-month periods while inserting either 0.9% unpreserved unit-dose saline (control) or multidose OPTI-FREE® RepleniSH™ (test RWD). Subjective comfort and symptoms were assessed after 2 and 4 weeks with each product. After 1 month of wear with each product, lenses were collected and analyzed in the laboratory for total protein, total lysozyme, and percentage of denatured lysozyme. Results. Symptoms of dryness and comfort varied across the day regardless of drop type (p < 0.001) with dryness being maximal on waking, least in the middle of the day, and increased towards the evening. The test RWD provided greater comfort on insertion (p = 0.02), better visual quality (p < 0.01), and less mucous discharge on waking (p = 0.02) than the control product. Lysozyme deposition was significantly reduced after the use of the test RWD as compared to saline (0.73 ± 0.5 &mgr;g/lens vs. 1.14 ± 0.7 &mgr;g/lens; p < 0.001) as was total protein deposition (1.17 ± 0.7 &mgr;g/lens vs. 1.86 ± 0.8 &mgr;g/lens; p < 0.001). Lysozyme denaturation was also reduced with the use of the test RWD compared with the control (76 ± 10% vs. 85 ± 7%; p < 0.01). Conclusions. The use of a RWD containing surface active agents provided greater subjective satisfaction, reduced lysozyme and total protein deposition, and reduced denatured lysozyme than a RWD containing saline alone.

The Impact of Tear Film Components on In Vitro Lipid Uptake

Purpose. To analyze the influence of various tear film components on in vitro deposition of two lipids (cholesterol and phosphatidylcholine) on three contact lens materials. Methods. Etafilcon A, balafilcon A, and senofilcon A were incubated in four different incubation solutions for 3 or 14 days: an artificial tear solution containing lipids and proteins, a protein tear solution containing proteins and the lipid of interest, a lipid tear solution containing lipids and no proteins, and a single lipid tear solution containing the lipid of interest only. Each incubation solution contained one of the two radiolabeled lipids: 14C-cholesterol (C) or 14C-phosphatidylcholine (PC). After soaking, lenses were removed from the incubation solution, the lipids were extracted and quantified using a beta counter, and masses of lipid were calculated using standard calibration curves. Results. This experiment examined several different parameters influencing lipid deposition on contact lenses, including lens material, length of incubation, and the composition of the incubation solution. Overall, lipid deposited differently on different lens materials (p < 0.0005), with the order of deposition most commonly being balafilcon > senofilcon > etafilcon. Incubation solution had a large impact on how much lipid was deposited (p < 0.00001), although cholesterol and phosphatidylcholine demonstrated different deposition patterns. Lipid deposition after 14 days of incubation was consistently greater than after 3 days (p < 0.02). Conclusions. This in vitro study demonstrates that C and PC deposition are cumulative over time and that silicone hydrogel materials deposit more lipid than group IV conventional hydrogel materials. It also clearly demonstrates that deposition of C and PC is influenced by the composition of the incubation solution and that in vitro models must use more physiologically relevant incubation solutions that mimic the natural tear film if in vitro data is to be extrapolated to the in vivo situation.

Quantification of non-polar lipid deposits on senofilcon a contact lenses.

Purpose. To quantify non-polar lipids deposited on senofilcon A silicone hydrogel contact lenses (J&J Acuvue OASYS) when disinfected with a no-rub one-step hydrogen peroxide system (CIBA Vision ClearCare) and a care system preserved with Polyquad & Aldox (Alcon OPTI-FREE RepleniSH). Methods. Thirty existing soft lens wearers symptomatic of dryness were enrolled into a 4-week prospective, randomized, bilateral eye (lens type), cross-over (care regimen), daily wear, double masked study. Subjects were refitted with senofilcon A lenses, which were replaced biweekly. During each period of wear, participants used either the peroxide or preserved system. After each period of wear, lenses were collected and lipid was extracted using 1.5 ml of a 2:1 chloroform:methanol solution for 3 h at 37°C. Lens extracts were analyzed for non-polar lipids [cholesterol oleate (CO), cholesterol, oleic acid (OA), triolein, and OA methyl ester] using normal phase high-performance liquid chromatography. Results. The total lipid (sum of CO and cholesterol) detected was 34 ± 28 &mgr;g/lens for the peroxide-based system and 22 ± 21 &mgr;g/lens for the system preserved with Polyquad and Aldox (p = 0.029). Although there was no difference between products for cholesterol (1.4 vs. 1.3 &mgr;g/lens; p = 0.50), use of a system preserved with Polyquad and Aldox resulted in significantly less deposited CO (33 ± 28 vs. 21 ± 20 &mgr;g/lens; p = 0.033). Approximately, 95% of the detectable lipid deposited on the material was CO, followed by cholesterol. OA and triolein contributed <1% of the total lipid and no OA methyl ester was found on any of the lenses. Conclusions. A care system preserved with Polyquad and Aldox removed higher amounts of CO from senofilcon A contact lenses used for 2 weeks than a peroxide-based system, in soft lens wearers who were symptomatic of dry eye.

The impact of intermittent air exposure on lipid deposition.

Purpose. To analyze the impact of intermittent air exposure on the in vitro deposition of two radioactive lipids on various contact lens (CL) materials, using a custom-designed model blink cell. Methods. Six different CL materials (balafilcon A, lotrafilcon B, comfilcon A, senofilcon A, etafilcon A, and omafilcon A) were mounted on the model blink cell pistons, which cycled the lenses in and out of a complex artificial tear solution (ATS) that contained a trace amount of 14C-cholesterol or 14C-phosphatidylcholine. For the short-term experiment, air-exposed lenses were continuously cycled in and out of the ATS for 10 h. Longer term incubations for 6 days were tested with lotrafilcon B and balafilcon A materials incubated in 14C-cholesterol ATS. The air-exposed CLs were cycled for 14 h then submerged for 10 h each day. For both experiments, the control lenses were submerged for the entire test period. After incubation, lenses were processed, and deposited masses were quantified. Results. Exposure to air resulted in increased amounts of cholesterol deposited by 1.6 to 4.3 fold on omafilcon A, balafilcon A, comfilcon A, and senofilcon A (p ⩽ 0.03) compared with submerged lenses. No differences in deposition were observed for etafilcon A and lotrafilcon B (p > 0.05). The longer term incubation of lotrafilcon B and balafilcon A showed statistically significant increases in cholesterol deposition for both air-exposed lens materials (p < 0.02) with the increase in deposition 1.8× and 2.8×, respectively. For phosphatidylcholine, all air-exposed lenses had increased masses of deposition. These deposits were statistically greater by 1.1 to 1.6 times for omafilcon A, comfilcon A, lotrafilcon B, and senofilcon A (p < 0.04), but not statistically different for etafilcon A or balafilcon A (p > 0.05). Conclusions. This study found that lipid deposition profiles are CL material dependent and that intermittent air exposure can influence the mass of lipid deposited.

Factors that influence in vitro cholesterol deposition on contact lenses.

Purpose The purpose of this study was to analyze the impact that incubation time, lipid concentration, and solution replenishment have on silicone hydrogel (SiHy) and conventional hydrogel (CH) contact lens cholesterol deposition via in vitro radiochemical experiments. Methods Four SiHy (senofilcon A, lotrafilcon B, comfilcon A, balafilcon A) and two CH (etafilcon A and omafilcon A) contact lenses were incubated in an artificial tear solution (ATS) that contained major tear film proteins, lipids, salts, salts, and a trace amount of radioactive 14C-cholesterol. Lenses were incubated for various incubation times (1, 3, 7, 14, or 28 days), with three concentrations of lipid (0.5×, 1×, 2× tear film concentration) and with or without solution replenishment to assess each variable’s impact on cholesterol deposition. After incubation, the lenses were extracted using 2:1 chloroform:methanol, extracts were analyzed in a beta counter and masses (micrograms per lens) were extrapolated from standard curves. Results Within the SiHy materials, balafilcon A deposited the greatest amount of cholesterol (p < 0.001) and lotrafilcon B the lowest (p < 0.001). The CH lens materials showed significantly lower uptake amounts than any of the SiHy lens materials (p < 0.001). The uptake of cholesterol ranged from 0.01 ± 0.01 &mgr;g/lens to 3.22 ± 0.34 &mgr;g/lens for all lens materials. Kinetic uptake of cholesterol was shown to be continuous throughout the 28 days of incubation without plateau (p < 0.001), and varying the lipid concentration did impact the resulting cholesterol deposition (p < 0.001). Replenishing the ATS every other day also affected cholesterol deposition throughout the experiment. Overall, the deposition pattern was 2× > replenishing > 1× > 0.5×. Conclusions Overall, SiHy lenses deposit significantly more cholesterol than CH lens materials, and the mass of lipid deposited is dependent on the contact lens material, length of incubation, concentration of lipids in the ATS, and the replenishment of ATS.

Using an In Vitro Model of Lipid Deposition to Assess the Efficiency of Hydrogen Peroxide Solutions to Remove Lipid from Various Contact Lens Materials

Purpose: To test the ability of two commercially available hydrogen peroxide disinfection solutions, one containing a surfactant and one without, to remove lipid from various contact lens materials using in vitro radiochemical experiments. Methods: Etafilcon A, senofilcon A and balafilcon A contact lens materials were incubated in an artificial tear solution (ATS) containing a mixture of lipids, proteins, mucin and either 14C-cholesterol or 14C-phosphatidylcholine for 8 h. Following incubation, the lenses were removed, rinsed, and placed for 16 h in either a surfactant-containing peroxide solution (ClearCare®), a peroxide solution devoid of a surfactant (AOSept®) or stored without solution (control). This process was repeated every day for 1 week. The lenses were extracted with a previously optimized extraction protocol, evaporated, re-suspended, fluor added and counted for their radioactive signals. Masses of lipids deposited were calculated based on standard calibration curves, the disinfection solutions were compared and repeated measures ANOVA and post hoc statistical analysis was completed using Statistica 9. Results: The results of this experiment found that daily disinfection with hydrogen peroxide solutions reduced the amount of cholesterol and phosphatidylcholine deposited on the three contact lens materials examined, however in many cases the reduction in deposition was less than 15% when compared to the control. Disinfection with the solution containing the surfactant (ClearCare), resulted in the least deposited cholesterol and phosphatidylcholine for all materials, however not all of the comparisons were statistically significant. Conclusions: Overall, ClearCare hydrogen peroxide disinfection solution containing Pluronic 17R4 removed the most lipid from lenses when compared to the non-surfactant containing AOSept or the control, for both lipids and all lens materials. However, the differences found were quite small at times and whether these differences are clinically significant are yet to be determined.

Atomic force microscopy and Langmuir–Blodgett monolayer technique to assess contact lens deposits and human meibum extracts

PURPOSE The purpose of this exploratory study was to investigate the differences in meibomian gland secretions, contact lens (CL) lipid extracts, and CL surface topography between participants with and without meibomian gland dysfunction (MGD). METHODS Meibum study: Meibum was collected from all participants and studied via Langmuir-Blodgett (LB) deposition with subsequent Atomic Force Microscopy (AFM) visualization and surface roughness analysis. CL Study: Participants with and without MGD wore both etafilcon A and balafilcon A CLs in two different phases. CL lipid deposits were extracted and analyzed using pressure-area isotherms with the LB trough and CL surface topographies and roughness values were visualized using AFM. RESULTS Meibum study: Non-MGD participant meibum samples showed larger, circular aggregates with lower surface roughness, whereas meibum samples from participants with MGD showed more lipid aggregates, greater size variability and higher surface roughness. CL Study: Worn CLs from participants with MGD had a few large tear film deposits with lower surface roughness, whereas non-MGD participant-worn lenses had many small lens deposits with higher surface roughness. Balafilcon A pore depths were shallower in MGD participant worn lenses when compared to non-MGD participant lenses. Isotherms of CL lipid extracts from MGD and non-MGD participants showed a seamless rise in surface pressure as area decreased; however, extracts from the two different lens materials produced different isotherms. CONCLUSIONS MGD and non-MGD participant-worn CL deposition were found to differ in type, amount, and pattern of lens deposits. Lipids from MGD participants deposited irregularly whereas lipids from non-MGD participants showed more uniformity.

Comparative study of lens solutions' ability to remove tear constituents.

Purpose The purpose of this study was to use atomic force microscopy to compare and characterize the cleaning abilities of a hydrogen peroxide–based system (HPS) and a polyhexamethylene biguanide–containing multipurpose solution (MPS) at removing in vitro deposited tear film constituents, as well as to determine deposition patterns on various silicone hydrogel contact lenses. Methods Silicone hydrogel materials—balafilcon A (BA), lotrafilcon B (LB), and senofilcon A (SA)—were incubated for 1 week in an artificial tear solution (ATS) containing representative lipids, proteins, and salts from the tear film. Atomic force microscopy was used to resolve each lens before and after being cleaned overnight in HPS or MPS. Atomic force microscopy was used again to resolve HPS/MPS-cleaned lenses, which were reincubated in fresh ATS for 1 week, before and after an overnight clean in their respective cleaning solution. Results Atomic force microscopy imaging was able to characterize lens deposits with high resolution. Lenses incubated in ATS revealed distinct differences in their deposition pattern across lens materials. The surface of BA contained about 20-nm-high deposits, whereas deposit heights up to 150 nm completely occluded the surface of SA. Lotrafilcon B lenses revealed clusters of deposits up to 90 nm. The use of either lens solution left trace amounts of tear film constituents, although components from the MPS were seen adsorbed onto the surface after cleaning. Surface roughness (Ra) measurements revealed a significant difference between ATS-incubated and HPS/MPS-cleaned SA and LB lenses (p < 0.05). Ra between first incubated and HPS/MPS-cleaned reincubated SA and LB was also significant (p < 0.05). Conclusions Unique variations in ATS deposition patterns were seen between lenses with atomic force microscopy. The application of both HPS and MPS removed most visible surface deposits.

A Pilot Study Investigating Clinical Responses and Biological Pathways of Azelastine/Fluticasone in Nonallergic Vasomotor Rhinitis before and after Cold Dry Air Provocation

Background: Nonallergic vasomotor rhinitis (NAVMR) has been considered a diagnosis by exclusion due to unknown mechanisms or lack of diagnostic biomarkers. Methods: To determine clinical responses and biological pathways in NAVMR subjects challenged to cold dry air (CDA) in an environmental exposure chamber (EEC) pre- and posttreatment with azelastine/fluticasone (AzeFlu), 30 NAVMR subjects, prescreened for CDA-induced symptoms (approx. 14°C, <15% relative humidity, ×1 h) were randomized to treatment with AzeFlu (n = 20) or placebo (n = 10) for 2 weeks. Total nasal symptoms scores, minimum cross-sectional area, cough, and conjunctival redness were recorded at visit 1 (pretreatment) and visit 2 (posttreatment) before, during, and after CDA challenge. At both visits, nasal lavage fluid (NLF) and nasal scrapings (NS) were collected pre- and post-CDA challenge. Substance P, neurokinin-A, and calcitonin gene-related peptide concentrations in NLF were analyzed pre- and postchallenge at each visit. Their relationship with CDA-induced symptoms was determined by statistical analysis. MicroRNA sequencing from NS determined differentially expressed miRNA between the treatment groups post-CDA challenge at each visit. Results: The minimum cross-sectional area (p < 0.05), cough count (p < 0.05), and substance P (p < 0.01) improved posttreatment with AzeFlu versus placebo. Gene targets for differentially expressed miRNAs at visit 1 were enriched for biological pathways regulating epithelial ciliogenesis and cell integrity that were modified in the AzeFlu-treated group versus placebo posttreatment. Conclusions: This study demonstrated the feasibility of an EEC model to investigate CDA-induced clinical responses and pathobiology in NAVMR subjects pre- and posttreatment with AzeFlu. NAVMR disease mechanisms for other nonallergic triggers can be investigated similarly.