How 1,n-Bis(3-alkylimidazolium-1-yl) Alkane Interacts with the Phospholipid Membrane and Impacts the Toxicity of Dicationic Ionic Liquids.

Ionic liquids based on doubly charged cations, often termed dicationic ionic liquids (DILs), offer robust physicochemical properties and low toxicity than conventional monocationic ionic liquids. In this design-based study, we used solid-state NMR spectroscopy to provide the interaction mechanism of two DILs, 1,n-bis(3-alkylimidazolium-1-yl) alkane dibromide ([C2n(C7-nIM)2]2+·2Br-, n = 1, 6), with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG) phospholipid membranes, to explain the low toxicity of DILs toward HeLa, Escherichia coli, Bacillus subtilis, and Saccharomyces cerevisiae cell lines. Dications with a short linker and long terminal chains cause substantial perturbation to the bilayer structure, making them more membrane permeabilizing, as shown by fluorescence-based dye leakage assays. The structural perturbation is even higher than [C12(MIM)]+ monocations, which carry a single 12-carbon long chain and exhibit a much higher membrane affinity, permeability, and cytotoxicity. These structural details are a crucial contribution to the design strategies aimed at harnessing the biological activity of ionic liquids.

Amphiphilic ionic liquid induced fusion of phospholipid liposomes.

We have investigated the impact of increasing concentration of imidazolium-based ionic liquids ([CnMIM]+[Br]-) on the structural integrity of large unilamellar vesicles (LUVs) made of pure phosphatidylcholine (PC) and phosphatidylglycerol (PG) lipids. Calcein based dye leakage assays were used to monitor the permeability of LUVs in the presence of ionic liquids. As the ionic liquid concentration approaches the critical micelle value, vesicle fusion occurs resulting in unexpected quenching which is accompanied by rapid dye leakage due to the formation of transiently lived fusion-holes. Vesicle fusion is confirmed using dynamic light scattering based size measurements and fluorescence based lipid mixing assays. 1H-1H NOESY measurements using solid-state NMR spectroscopy were performed to obtain insights into the fusion mechanism. While POPC LUVs are more prone to membrane fusion, the overall extent of fusion is higher in POPG LUVs. Ionic liquid induced splaying of phospholipid chains is crucial for overcoming the hydration barrier between the merging bilayers.

Effect of the Alkyl Chain Length of Amphiphilic Ionic Liquids on the Structure and Dynamics of Model Lipid Membranes.

We compare the biophysical and structural aspects of the interaction of amphiphilic ionic liquids containing 1-alkyl-3-methylimidazolium cation ([CnMIM]+, n = 8, 12, or 16) with membranes composed of zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) or anionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-rac-glycerol (POPG). Liposome affinity and permeabilization were determined using ζ-potential and fluorescence studies, correlated with the cytoxicity of [CnMIM]+Br- toward HeLa cell lines. Membrane affinity is strongest in the case of [C16MIM]+Br- followed by [C12MIM]+Br- and [C8MIM]+Br- for both membranes, and trends remained the same in the case of membrane permeability and cytotoxicity. Solid-state NMR spectroscopy was used to localize [CnMIM]+ inside the lipid bilayers and to study their impact on the head group and acyl chain structures and dynamics of the lipid molecules. The charged ring moiety of the [CnMIM]+ is localized in the lipid-water interface of the membranes irrespective of the chain length and membrane surface charge. While [C8MIM]+ binds the membrane most weakly, it induces the largest disorder in the lipid chain region. A lack of fast flip-flop motions of the amphiphiles in the case of long chain [C16MIM]+ is suggested to render the membrane unstable, which increases its permeability. Between the lipid molecules, the POPC membrane incurs larger disorder in lipid chain packing upon insertion of [CnMIM]+ molecules. The study provides structural details of the impact of increasing chain lengths in [CnMIM]+ on the structural properties of lipid bilayers.

Long-term Retrospective Study based on Implant Success Rate in Patients with Risk Factor: 15-year Follow-up.

AIM: The purpose of this retrospective study is to assess implant success rates with various risk factors. MATERIALS AND METHODS: Two hundred patients with a total of 650 implants were selected. Risk factors, such as smoking, antidepressants, bruxism, diabetes, and bone augmentation procedures were considered, and patients were followed up for a period of 8 to 15 years. RESULTS: Of 650 implants placed, the success rate was 88%, i.e., a total of 572 implants were successful. A total of 78 implants were considered failure; and out of 78, twenty implants were surgically removed. CONCLUSION: Based on this study's results, it is concluded that risk factors, such as smoking, bruxism, diabetes, and bone augmentation play an important role in success rate of dental implants. CLINICAL SIGNIFICANCE: Several factors, such as bruxism, diabetes, and supporting bone can play an important role in dental implant success.

Impact of Vitiligo on Quality of Life in Patients of Skin of Color and Its Correlation With Clinical Severity Assessment Scores Utilizing Disease Specific Scores: A Cross-Sectional Study.

INTRODUCTION: Assessment of disease severity of vitiligo is exigent as it is a psychosomatic ailment. VIDA (vitiligo disease activity score) and VASI (vitiligo area severity index) were previously used for this evaluation. Recently, the introduction of two vitiligo specific tools, vitiligo impact scale (VIS)-22 and Vitiligo Quality of Life Index (VitiQoL) has aided in assessing the quality of life (QOL) in a pertinent manner. OBJECTIVES: To measure the QOL in vitiligo using disease specific indices (VitiQoL and VIS-22), to assess their relationship with disease severity (VASI and VIDA) and to determine the correlation between QOL scores (VIS-22 and VitiQoL). METHODS: This observational cross-sectional study included 195 patients with vitiligo, and their disease severity was calculated using VASI and VIDA scoring. Patients were asked to fill questionnaires for assessing the QOL using validated tools i.e. VIS-22 and VitiQoL. RESULTS: Significant correlation was demonstrated between both QOL scores and VASI score (P value 0.001) with slightly higher values for VitiQoL (r = 0.824) than with VIS 22 (r = 0.693). Both scores exhibited a significant association with VIDA score (P value < 0.001). Moreover, statistically significant correlation was found between VIS-22 and VitiQoL, thereby proving the concordance between these scores. CONCLUSIONS: The study infers that QOL seemed to be remarkably dependent on the clinical severity scores and that higher disease activity corresponds to poorer QOL. It is imperative to precisely assess burden of vitiligo and the impairments caused by it in order to aid multi-modality management and allow more standardized research.

Role of Cholesterol in Interaction of Ionic Liquids with Model Lipid Membranes and Associated Permeability.

In this work, we explored how the amount of cholesterol in the lipid membrane composed of phosphatidylcholine (POPC) or phosphatidylglycerol (POPG) affects the interaction with 1-dodecyl-3-methylimidazolium bromide ([C12MIM]+Br-) ionic liquids using various biophysical techniques. On interacting with the membrane, [C12MIM]+Br- leads to enhanced membrane permeability and induces membrane fusion, leading to an increase in vesicle size. The 2H-based solid-state NMR investigations of cholesterol-containing lipid membranes reveal that [C12MIM]+Br- decreases the lipid chain order parameters and counteracts the lipid condensation effect of cholesterol to some extent. Therefore, as the amount of cholesterol in the membrane increases, the membrane effect of [C12MIM]+Br- decreases. The effect of [C12MIM]+Br- on the membrane properties is more pronounced for POPC compared to that of POPG membranes. This suggests a dependence of these effects on the electrostatic interactions, indicating that the influence of [C12MIM]+Br- varies based on the lipid composition. The findings suggest that the presence of cholesterol can modulate the effect of [C12MIM]+Br- on membrane properties, with variations observed between POPC and POPG membranes, highlighting the importance of lipid composition. In short, this study provides insights into the intricate interplay between cholesterol, the lipid membrane, and the ionic liquid [C12MIM]+Br-.

Impact of Lipid Ratio on the Permeability of Mixed Phosphatidylcholine/Phosphatidylglycerol Membranes in the Presence of 1-Dodecyl-3-methylimidazolium Bromide Ionic Liquid.

We have studied the impact of the lipid ratio on the membrane permeability of mixed phosphatidylcholine (POPC)/phosphatidylglycerol (POPG) membranes induced by 1-dodecyl-3-methylimidazolium bromide ([C12MIM]+Br-) ionic liquid by evaluating the role of affinity and architecture of the phospholipid bilayer. Nine different model membranes composed of negatively charged POPG and zwitterionic POPC lipids mixed in molar ratios of 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, and 1:9 have been studied. The membrane permeability of each composition has been evaluated using fluorescence-based dye leakage assays. Despite having the highest membrane affinity, POPG-rich membranes doped with 10 and 20 mol % POPC are found to be the least permeable. 31P- and 2H-based solid-state NMR investigations reveal that the minor POPC component is homogeneously dispersed in the PG/PC (8:2) membrane. In contrast, the lipids seem to be segregated into POPG- and POPC-rich domains in the complementary PG/PC (2:8) composition. Although [C12MIM]+ cations have a stronger interaction with the POPG component in the mixed membranes, their insertion has a limited impact on the overall structure and dynamics of the PG/PC (8:2) composition.

Relationship between fracture toughness and fractal dimensional increment in two types of dental glass-ceramics with different fracture surface roughness.

OBJECTIVES: Previous studies have reported the fractal dimensional increment of glass-ceramic fracture surfaces. The objective of this study was to determine the relationship between fracture toughness and fractal dimensional increment of two dental glass-ceramics with different volume fraction of crystals and different fracture surface roughness. METHODS: Bar-shaped specimens were prepared from lithium disilicate (LDS) and nanofluorapatite (NFA) glass-ceramics. One face of each specimen was indented using a Knoop diamond at 25 N (LDS) or 10 N (NFA) followed by loading in 4-point, or 3-point flexure, respectively, until failure. Fracture toughness (Kc) was calculated using the surface crack in flexure (SCF) technique (ASTM C1421). Epoxy replicas of the fracture surfaces were scanned using the atomic force microscope (AFM) followed by noise filtering. The FRACTALS software was used to determine the fractal dimensional increment (D*) by the Minkowski cover algorithm. RESULTS: Median (25%, 75% quartiles) fracture toughness of LDS bars were 1.62 (1.59, 1.69) MPa m1/2 and NFA bars were 0.68 (0.66, 0.74) MPa m1/2, respectively. The median fractal dimension (D) value (25%, 75% quartiles) before noise filtering were 2.16 (2.15, 2.17) and after noise filtering were 2.14 (2.14, 2.15) for LDS and before noise filtering were 2.29 (2.21, 2.38) and after noise filtering were 2.17 (2.17, 2.18) for NFA. Median (25%, 75% quartiles) surface roughness (Ra) before noise filtering were 139 (119, 188) nm and after noise filtering were 137 (118, 187) nm for LDS and before noise filtering were 7 (6, 15) nm and after noise filtering were 7 (6, 15) nm for NFA. SIGNIFICANCE: Noise filtering successfully eliminated noise from the material with smooth fracture surfaces (NFA), decreasing the measured fractal dimension. The NFA data fit a Kc vs. D*1/2 statistical model for fused silica previously tested using a similar technique. The equation relating fracture toughness to the fractal dimension was modified, accounting for the toughening mechanisms. Fractal analysis with noise filtering can be used to estimate the fracture toughness of dental glass-ceramics that do not exhibit crack bridging.

Cytotoxicity and Membrane Permeability of Double-Chained 1,3-Dialkylimidazolium Cations in Ionic Liquids.

We have evaluated ionic liquids based on double-chained 1-alkyl-3-octylimidazolium cations ([CnC8IM]+, n = 2, 4, 6, 8, 10, 12) for their cytotoxicity toward various cell lines. The toxicity of ionic liquids was correlated to their ability to partition into and permeabilize phosphocholine (POPC)- or phosphoglycerol (POPG)-based large unilamellar vesicles. Membrane partitioning of ionic liquids was assessed using the ζ-potential measurements, and membrane permeability was determined using fluorescence-based dye leakage assays. Both cytotoxicity and membrane permeability of these ILs were found to increase in a sigmoidal fashion with increasing chain length on the N1 atom (n in [CnC8IM]+) cations. These results were compared with those for ionic liquids based on single-chained 1-alkyl-3-methylimidazolium cations ([Cn+8C1IM]+), carrying a similar number of carbon atoms but as a single alkyl chain. Our studies show that ionic liquids containing double-chained cations are relatively less cytotoxic and membrane-permeabilizing than the cations bearing a single long alkyl chain.

Role of cationic head-group in cytotoxicity of ionic liquids: Probing changes in bilayer architecture using solid-state NMR spectroscopy.

The effect of cationic head-group of ionic liquid on the structure and dynamics of phospholipid bilayer was studied to provide insights into the mechanism of ionic liquid-membrane interaction. The effect was observed using six ionic liquids containing benzimidazolium, imidazolium, pyrrolidinium, piperidinium, ammonium, and morpholinium based amphiphilic cations carrying a dodecyl alkyl chain. Unilamellar and multilamellar vesicles composed of zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were used. Permeability of POPC bilayer was found to have a strong dependence on ionic liquid head-group structure. To probe the structural details of interaction, 31P and 2H based solid-state NMR measurements were performed. The cations differed in terms of their effect on the orientation and disorder in the phosphocholine moiety in lipid head-group as revealed by chemical shift anisotropy of 31P. Cations carrying an unshielded charge like benzimidazolium, imidazolium, and ammonium result in strong reorientation of phosphocholine moiety in lipid head-group. Large sized cations like benzimidazolium and piperidinium result in enhanced lipid chain dynamics as revealed by order parameter calculations of deuterated lipid chains. Relatively polar head-group of morpholinium cation neither impacts the phospholipid head-group nor chain packing. Our results suggest that there exists a direct correlation between ionic liquid head-group induced structural changes in bilayer and their ability to permeabilize/disrupt the membrane and be cytotoxic.

Amphiphilic Ionic Liquid-Induced Membrane Permeabilization: Binding Is Not Enough.

The interaction of amphiphilic ionic liquids containing an 1-alkyl-3-methylimidazolium cation ([C12MIM]+), which shows acute cytotoxicity toward marine and bacterial life, with zwitterionic 1-palmitoyl-2-oleoyl- sn-glycero-3-phospho-choline (POPC) and anionic 1-palmitoyl-2-oleoyl- sn-glycero-3-phospho- rac-glycerol (POPG) membranes was investigated. Biophysical parameters of this interaction were quantified by fluorescence spectroscopy, isothermal titration calorimeter, and solution-state NMR measurements. [C12MIM]+ inserts into the membrane and induces vesicle leakage at relatively low concentration (<1 mM). Zwitterionic POPC membranes are more leakage-prone, but the binding of [C12MIM]+ cations is stronger to anionic POPG membranes. A higher rate of exchange of membrane-bound [C12MIM]+ is suspected to play a key role in membrane leakage. Furthermore, solid-state NMR spectroscopy was employed to determine the location of [C12MIM]+ in lipid membranes and its impact on the structure and dynamics of the bilayer. The study provides a molecular understanding of the membrane permeabilizing effect of the [C12MIM]+ mediated by its detergent-like structure.

Sickle cell patient with an acute chest syndrome and a negative chest X-ray: potential role of the ventilation and perfusion (V/Q) lung scan.

Acute chest syndrome (ACS) in sickle cell disease is caused by thromboemboli in the pulmonary vasculature. The diagnostic criteria include the presence of pulmonary infiltrate(s) on chest x-ray. This case report suggests that a V/Q scan may play a diagnostic role in sickle cell patients with symptoms of ACS and a negative chest x-ray.

Potential role of the ventilation and perfusion (V/Q) lung scan in the diagnosis of acute chest syndrome in adults with sickle cell disease.

The current criteria for diagnosing ACS are chest pain and presence of a new infiltrate on the chest radiograph (CXR). This study was designed to evaluate the role of ventilation and perfusion (V/Q) scan to assist in the early diagnosis of ACS. An abnormal V/Q scan was associated with a diagnosis of ACS that reached a statistical significance (P < 0.038). The sensitivity and specificity were found to be 60% and 100%, respectively. We conclude that V/Q scan may play a role in the early diagnosis of ACS.