Lamellar Organization Research Articles

Stable and Fluid Multilayer Phospholipid–Silica Thin Films: Mimicking Active Multi-lamellar Biological Assemblies

Phospholipid-based nanomaterials are of interest in several applications including drug delivery, sensing, energy harvesting, and as model systems in basic research. However, a general challenge in creating functional hybrid biomaterials from phospholipid assemblies is their fragility, instability in air, insolubility in water, and the difficulty of integrating them into useful composites that retain or enhance the properties of interest, therefore limiting there use in integrated devices. We document the synthesis and characterization of highly ordered and stable phospholipid-silica thin films that resemble multilamellar architectures present in nature such as the myelin sheath. We have used a near room temperature chemical vapor deposition method to synthesize these robust functional materials. Highly ordered lipid films are exposed to vapors of silica precursor resulting in the formation of nanostructured hybrid assemblies. This process is simple, scalable, and offers advantages such as exclusion of ethanol and no (or minimal) need for exposure to mineral acids, which are generally required in conventional sol-gel synthesis strategies. The structure of the phospholipid-silica assemblies can be tuned to either lamellar or hexagonal organization depending on the synthesis conditions. The phospholipid-silica films exhibit long-term structural stability in air as well as when placed in aqueous solutions and maintain their fluidity under aqueous or humid conditions. This platform provides a model for robust implementation of phospholipid multilayers and a means toward future applications of functional phospholipid supramolecular assemblies in device integration.

Protective effect of Spirulina platensis enriched in phenolic compounds against hepatotoxicity induced by CCl4

Phenolic compounds make up the major secondary metabolites with high pharmaceutical potential. Microalgae were reported to contain low amounts of phenolic compounds. The present study aimed to investigate the hepatoprotective potential of biomass of Spirulina platensis enriched in phenolic compounds. The protective effects of the biomass of S. platensis with low amounts of phenolics (SP1) and with high amounts of phenolics (SP2) against CCl4-induced acute hepatotoxicity were evaluated in rats. The increased levels of ALT, AST and MDA along with decreased activities of SOD and CAT were significantly (p<0.01) ameliorated by SP2. Histological examinations revealed that SP2 was more potent than SP1 in protecting the liver from toxic injury of CCl4 and preserving the hepatocyte ultrastructure. The lesions including necrosis, lymphocyte infiltration, ballooning degeneration and hepatocyte injury as irregular lamellar organisation, dilations in endoplasmic reticulums and the presence of great number of cytoplasmic vacuolization were healed by SP2.

Visualisation of methacrylate‐embedded human bone sections by infrared nanoscopy

A recently developed ultra-resolving near-field infrared nanoscope is applied to investigate methyl methacrylate embedded, un-decalcified human bone sections. Results show detail at a resolution of 30 nm. Specific contrasting of mineral components is enabled by choosing an appropriate infrared wavelength, here 9.47 μm, in the phosphate vibrational band. The method is surface-sensitive, probing to a depth of about 30 nm into the surface. The obtained infrared images are presented in direct comparison with optical and electron micrographs of the identical specimen. Lamellar bone organization, peri-cellular mineral deposition, and regional differences in mineral content are clearly detectable. Individual fibrils are resolved. - Infrared nanoscopy requires just standard hard tissue preparation techniques combined with section surface polishing. It can be integrated into existing laboratory environments without impeding subsequent routine staining and evaluation methods.

Regional variations in the nonlinearity and anisotropy of bovine aortic elastin

Arterial walls have a regular and lamellar organization of elastin present as concentric fenestrated networks in the media. In contrast, elastin networks are longitudinally oriented in layers adjacent to the media. In a previous model exploring the biomechanics of arterial elastin, we had proposed a microstructurally motivated strain energy function modeled using orthotropic material symmetry. Using mechanical experiments, we showed that the neo-Hookean term had a dominant contribution to the overall form of the strain energy function. In contrast, invariants corresponding to the two fiber families had smaller contributions. To extend these investigations, we use biaxial force-controlled experiments to quantify regional variations in the anisotropy and nonlinearity of elastin isolated from bovine aortic tissues proximal and distal to the heart. Results from this study show that tissue nonlinearity significantly increases distal to the heart as compared to proximally located regions ([Formula: see text]). Distally located samples also have a trend for increased anisotropy ([Formula: see text]), with the circumferential direction stiffer than the longitudinal, as compared to an isotropic and relatively linear response for proximally located elastin samples. These results are consistent with the underlying tissue histology from proximally located samples that had higher optical density ([Formula: see text]), fiber thickness ([Formula: see text]), and trend for lower tortuosity ([Formula: see text]) in elastin fibers as compared to the thinner and highly undulating elastin fibers isolated from distally located samples. Our studies suggest that it is important to consider elastin fiber orientations in investigations that use microstructure-based models to describe the contributions of elastin and collagen to arterial mechanics.

Inhibition of skin inflammation in mice by diclofenac in vesicular carriers: Liposomes, ethosomes and PEVs

Diclofenac-loaded phospholipid vesicles, namely conventional liposomes, ethosomes and PEVs (penetration enhancer-containing vesicles) were developed and their efficacy in TPA (phorbol ester) induced skin inflammation was examined. Vesicles were made from a cheap and unpurified mixture of phospholipids and diclofenac sodium; Transcutol P and propylene glycol were added to obtain PEVs, and ethanol to produce ethosomes. The structure and lamellar organization of the vesicle bilayer were investigated by transmission electron microscopy and small and wide angle X-ray scattering, as well as the main physico-chemical features. The formulations, along with a diclofenac solution and commercial Voltaren Emulgel, were tested in a comparative trial for anti-inflammatory efficacy on TPA-treated mice dorsal skin. Vesicles were around 100 nm, negatively charged, able to encapsulate diclofenac in good yields, and disclosed different lamellarity, as a function of the formulation composition. Vesicular formulations promoted drug accumulation and reduced the permeation. Administration of vesicular diclofenac on TPA-inflamed skin resulted in marked attenuation of oedema and leucocyte infiltration, especially using PEVs. Histology confirmed the effectiveness of vesicles, since they provided an amelioration of the tissual damage induced by TPA. The proposed approach based on vesicular nanocarriers may hold promising therapeutic value for treating a variety of inflammatory skin disorders.

Lamellar organization of discotic dimer enforced by steric manipulation

Dibenzo[a,c]phenazine dimer mesophases exhibited a lamellar assembly as demonstrated by XRD diffraction. This is a new successful molecular design to achieve a hierarchical supramolecular assembly for dimeric discotic liquid crystals. All the mesophases are readily frozen into a glassy state at room temperature.

Ridge Augmentation With Mineralized Block Allografts

Bone augmentation is frequently used to create sufficient bone volume for ideal implant placement. Severely resorbed ridges require extensive bone augmentation in the form of block allografts. A 3-dimensional graft technique has been developed to augment atrophic areas. This technique involves modifying the graft on a sterile prototype of the recipient site before the surgery. This article investigates the clinical and histological outcomes of ridge augmentation using this technique. Eight partially edentulous patients were recruited. Ridge augmentations were performed using block allografts, preadjusted, based on sterile prototypes of the recipient bed before the surgeries. After 8 months, 20 implants were inserted into the grafted sites. Eight bone cores were harvested for histological analysis. Highly vital and mineralized bone with lamellar organization was observed at the grafted sites. Having the ability to modify the allogeneic block grafts to fit the recipient sites before the surgery minimized the surgical time and risk of postoperative complications such as infection. In addition, the clinician could concentrate fully on achieving tension-free primary wound closure.

Effect of cream cooling rate and water content on butter microstructure during four weeks of storage

Crystallization, rheological properties and microstructure of butter with varying water content and subjected to different cooling rate were studied during four weeks of storage at 5 °C. Using small and large deformation rheology, the elastic modulus (G′) and Hencky strain at fracture was followed. When comparing samples with an equal water content, samples produced from fast cooled cream (7.5 °C/min) have a higher G′ compared to butter produced from slow cooled cream (0.4 °C/min), at day 1–7. However, no difference in G′ is observed as a function of time, even though the solid fat content increases. Increasing the water content from 20% to 32% decreases G′ at day 1–14, yet X-ray scattering and differential scanning calorimetry shows no difference in crystal polymorphism or crystallinity. After 21 days of storage, no difference in G′ is observed as a function of cream cooling rate or water content. For all samples, small angle X-ray scattering shows formation of 2L (41 Å) and 3L (57 Å) lamellar organization, while the wide angle spectra shows mainly β′-crystals (4.2 Å & 3.81 Å) together with traces of β (4.6 Å).

Structural variation of rice starch in response to temperature during microwave heating before gelatinisation

Although it is well known that structural variations occur in starch during gelatinisation, little is known about how the structure of starch changes at subgelatinisation levels. The objective of this study was to investigate structural variations of rice starch ascribed to the temperature during microwave heating. Rapid conduction heating was used to imitate the high microwave heating rate through oil bath, which was then compared with traditional conduction heating. Structural changes due to temperature increases were investigated using thermogravimetry and differential scanning calorimetry while the distinct lamellar organisation of starch was obtained through small-angle X-ray scattering. The results showed that the structure of starch responds non-monotonically to temperature rising before gelatinisation, which was also affected by heating rates. The samples treated by microwave and rapid conduction heating essentially underwent the same thermal property changes and the molecular vibration of the microwaves did influence the submicroscopic lamellar structure.

Perspectives on the history of research on starch Part V: On the conceptualization of amylopectin structure

AbstractStarch has been used over several millennia for a number of different applications. However, research on understanding this substance only spans about three centuries starting with Leeuwenhoek who observed it in 1716. This story of discovery of the molecular structure and architectural makeup of starch is chronicled in a series of six essays of which this is the fifth with a focus on the understanding of amylopectin structure. Research with a focus on the structure of amylopectin, the branched and major component in starch granules, started only in the 1940s when accurate techniques for the separation of amylose and amylopectin were developed. The understanding of amylopectin structure went hand in hand with research on starch granule crystallinity and lamellar organization. The discoveries of new enzymes involved in starch biosynthesis and degradation added to the understanding of amylopectin structure. Soon it became apparent that enzyme preparations used in this kind of research had to be of highest purity in order to achieve accurate results. The purification of debranching enzymes from bacterial sources, in combination with the new technique of gel‐permeation chromatography, revolutionized the understanding of the unit chain composition in amylopectin as being different from that of glycogen, and resulted in the proposal of the cluster structure of amylopectin.Please read the Editorial for more details.

Effect of amylose deposition on potato tuber starch granule architecture and dynamics as studied by lintnerization

The effect of amylose deposition on the amylopectin crystalline lamellar organization in potato starch granules was studied by mild acid, so-called lintnerization, of potato tuber starch transgenically engineered to deposit different levels of amylose. The starch granules were subjected to lintnerization at different temperatures (25, 35, and 45°C) and to two levels of solubilization, ∼ 45 and 80%. The rate of the lintnerization increased with temperature but was suppressed by amylose. The molecular size of the lintner dextrins increased with temperature, but this effect was suppressed by the presence of amylose. At high temperatures and low-amylose content, the degree of branches was high with the concomitant increase in size in the dextrins. A portion of the branches was resistant to debranching enzymes possibly due to specific structural formations. The effects of temperature suggested a unique granular architecture of potato starch, and a model showing the dependence of temperature on the dynamic arrangement of amylopectin and amylose in the crystalline and amorphous lamellae for the potato starch is suggested.

Characterization of the Solutol® HS15/water phase diagram and the impact of the Δ9-tetrahydrocannabinol solubilization

Here, the phase behavior of the commercial non-ionic surfactant Solutol® HS15 in water was investigated. The focus was on the evolution of the system nanostructure at low water content. Particularly, it was demonstrated that spherical micelles found in dilute surfactant solutions coalesce at a surfactant volume fraction close to 0.5. As consequence, a heterogeneous pseudo-binary mixture occurs. No liquid crystalline phases were detected even at the highest HS15 concentrations in water. Alteration of the micellar morphology induced by the addition of Δ9-tetrahydrocannabinol to the surfactant/water binary system was also investigated. It was found that the cannabinoid molecules become entrapped within the surfactant hydrophobic tails, thus increasing the surfactant effective packing parameter and inducing a radical change of the micelle shape. At sufficiently low water content (18–35wt.%), such alteration of the interfacial packing results in a lamellar organization of the surfactant molecules.

Structure and spectral sensitivity of photoreceptors of two anchovy species: Engraulis japonicus and Engraulis encrasicolus

The morphology, fine structure and spectral sensitivity of retinal photoreceptors of two anchovy species were investigated using light and electron microscopy and microspectrophotometry. Distinct regional specialisation of cones was observed. Long and short (bilobed) cones were observed in the horizontal retinal belt, including the nasal and temporal retinal zones. Only triple cones with two long lateral components, one small central component were observed in the dorsal and ventro-nasal retinal regions. The long cones presented various lamellar organisation patterns: (1) in parallel along the cell axis in the central retina, (2) oriented transversely at the base of the outer segment, and (3) tilted longitudinally while extending to the tip of the cone in the retinal periphery. In the short cones, the lamellae were always oriented along the cell axis, and their planes were perpendicular to the lamellae in the long cones, providing a structural basis for the detection of polarisation of incident light. The lamellae in all the outer segments of the triple cones are arranged perpendicular to the long cell axis. In both species, the long and short cones from the ventro-temporal retina were slender and more densely packed, and the outer segments of the long cones lay far more sclerad compared with the outer segments of the bifid cones. Microspectrophotometry revealed that in both species the lateral components of the triple cones displayed a maximum absorbance wavelength (λmax) of approximately 502nm, while the short central components were more shortwave sensitive (λmax=475nm). The λmax of all long and short cones in the ventro-temporal zone was 492nm, compared to 502nm in other retinal regions. Anchovies are unique among vertebrates in that they contain clear structural basis for both colour and polarisation vision in the same retina.

Synthesis and Thermotropic Liquid Crystallinity of Alkylammonium Salts Containing Azobenzene Mesogens

Alkylammonium salts have been successfully synthesized by reacting 1-iodoalkanes with an azobenzene mesogen of 1-[bis-(2-hydroxyethyl) amino]-6-(4-ethoxyazobenzene-4-oxy) hexane. They were found to mainly form thermotropic smectic liquid crystals with various textures under polarized light. Their phase transition temperatures vary with alkyl chains, and the ordering of lamellar organization decreases with increasing the length of alkyl tails. The as-synthesized liquid crystals may have potential application in the field of photoresponsive and nanostructured materials.

Molecular, mesoscopic and microscopic structure evolution during amylase digestion of maize starch granules

Cereal starch granules with high (>50%) amylose content are a promising source of nutritionally desirable resistant starch, i.e. starch that escapes digestion in the small intestine, but the structural features responsible are not fully understood. We report the effects of partial enzyme digestion of maize starch granules on amylopectin branch length profiles, double and single helix contents, gelatinisation properties, crystallinity and lamellar periodicity. Comparing results for three maize starches (27, 57, and 84% amylose) that differ in both structural features and amylase-sensitivity allows conclusions to be drawn concerning the rate-determining features operating under the digestion conditions used. All starches are found to be digested by a side-by-side mechanism in which there is no major preference during enzyme attack for amylopectin branch lengths, helix form, crystallinity or lamellar organisation. We conclude that the major factor controlling enzyme susceptibility is granule architecture, with shorter length scales not playing a major role as inferred from the largely invariant nature of numerous structural measures during the digestion process (XRD, NMR, SAXS, DSC, FACE). Results are consistent with digestion rates being controlled by restricted diffusion of enzymes within densely packed granular structures, with an effective surface area for enzyme attack determined by external dimensions (57 or 84% amylose – relatively slow) or internal channels and pores (27% amylose – relatively fast). Although the process of granule digestion is to a first approximation non-discriminatory with respect to structure at molecular and mesoscopic length scales, secondary effects noted include (i) partial crystallisation of V-type helices during digestion of 27% amylose starch, (ii) preferential hydrolysis of long amylopectin branches during the early stage hydrolysis of 27% and 57% but not 84% amylose starches, linked with disruption of lamellar repeating structure and (iii) partial B-type recrystallisation after prolonged enzyme incubation for 57% and 84% amylose starches but not 27% amylose starch.

A three-scale finite element investigation into the effects of tissue mineralisation and lamellar organisation in human cortical and trabecular bone

Bone is an exceptional material that is lightweight for efficient movement but also exhibits excellent strength and stiffness imparted by a composite material of organic proteins and mineral crystals that are intricately organised on many scales. Experimental and computational studies have sought to understand the role of bone composition and organisation in regulating the biomechanical behaviour of bone. However, due to the complex hierarchical arrangement of the constituent materials, the reported experimental values for the elastic modulus of trabecular and cortical tissue have conflicted greatly. Furthermore, finite element studies of bone have largely made the simplifying assumption that material behaviour was homogeneous or that tissue variability only occurred at the microscale, based on grey values from micro-CT scans. Thus, it remains that the precise role of nanoscale tissue constituents and microscale tissue organisation is not fully understood and more importantly that these have never been incorporated together to predict bone fracture or implant outcome in a multiscale finite element framework. In this paper, a three-scale finite element homogenisation scheme is presented which enables the prediction of homogenised effective properties of tissue level bone from its fundamental nanoscale constituents of hydroxyapatite mineral crystals and organic collagen proteins. Two independent homogenisation steps are performed on representative volume elements which describe the local morphological arrangement of both the nanostructural and microstructural levels. This three-scale homogenisation scheme predicts differences in the tissue level properties of bone as a function of mineral volume fraction, mineral aspect ratio and lamellar orientation. These parameters were chosen to lie within normal tissue ranges derived from experimental studies, and it was found that the predicted stiffness properties at the lamellar level correlate well with experimental nanoindentation results from cortical and trabecular bone. Furthermore, these studies show variations in mineral volume fraction, mineral crystal size and lamellar orientation could be responsible for previous discrepancies in experimental reports of tissue moduli. We propose that this novel multiscale modelling approach can provide a more accurate description of bone tissue properties in continuum/organ level finite element models by incorporating information regarding tissue structure and composition from advanced imaging techniques. This approach could thereby provide a preclinical tool to predict bone mechanics following prosthetic implantation or bone fracture during disease.

Novel Two-Component Gels of Cetylpyridinium Chloride and the Bola-amphiphile 6-Amino Caproic Acid: Phase Evolution and Mechanism of Gel Formation

A two-component gel resulting from the amphiphilic cationic surfactant cetylpyridinium chloride (CPC) in the presence of a structure-forming bola-amphiphilic additive 6-aminocaproic acid (6-ACA) was realized and investigated. At a critical 6 wt % of 1:1 CPC:6-ACA, the yellow colored gel resulted from a 3:1 v/v CHCl(3):H(2)O critical binary solvent composition. The mixed amphiphilic system formed a 1:1 complex with a binding constant ~0.83 × 10(4) M(-1). Phase evolution and mechanism of gelation in the mixed CPC:6-ACA system was unraveled upon investigating the gel microstructure, based on spectroscopic, microscopic, and small-angle X-ray scattering (SAXS) investigations. The gel assembled as a lamellar organization, maintaining a loosely interdigitated bilayer structure of CPC and 6-ACA molecules through predominant charge transfer, H-bonding, and hydrophobic and intercomplex interactions. The SAXS pattern indicated a semicrystalline form to be the stable phase with alternating crystalline and amorphous layers; a novel mode of gelation with a widely disparate semicrystalline form of the lamellar gel was thus indicated, where the lamellar structure was deduced from the interplanar spacings. A transition from low viscosity reverse micellar solution to a yellow rigid gel upon aging was thus comprehended. The mixed amphiphile in varying polarity organic solvents in the presence of water indicated the nonconducive nature of gelation in very highly polar solvents, methanol, and DMF or, in very low polarity solvents, such as, cyclohexane and carbon tetrachloride.

A Unique Bicellar Nanosystem Combining Two Effects on Stratum Corneum Lipids

In this work a new composition (dioleylphosphatidylcholine, DOPC, and dihexanoylphosphocholine, DHPC) is used to form the bicellar system and to evaluate their effect on stratum corneum (SC) lipids. Through this article, "bicellar system" will refer to a lipid binary system in which lipids are self-assembled forming different nanostructures. DOPC/DHPC system is characterized by dynamic light scattering and cryo-transmission electron microscopy showing two different nanostructures: unilamellar vesicles and tubular structures. In order to study the SC lipid organization attenuated total reflectance Fourier transform infrared spectroscopy, freeze-substitution applied to transmission electron microscopy and X-ray scattering are used. This work compares for the first time the use of two different X-ray scattering methods, transmission with synchrotron radiation and grazing incidence with conventional source, for skin studies. Our results indicate that vesicle-shaped structures remain adhered to the SC surface being unable to penetrate into the skin probably due to their large and voluminous size, while a proportion of structures could have interaction with SC lipids increasing the lamellar organization. Thus, the different nanostructures present in the system have different effects on SC lipids. The appropriate combination of both effects and the possibility to incorporate drugs offer a range of possibilities for the DOPC/DHPC system in development for skin care products.

Siliceous deep-sea sponge Monorhaphis chuni: A potential paleoclimate archive in ancient animals

The deep-sea sponge Monorhaphis chuni forms giant basal spicules, which can reach lengths of 3m; they represent the largest biogenic silica structures on Earth that is formed from an individual metazoan. The spicules offer a unique opportunity to record environmental change of past oceanic and climatic conditions. A giant spicule collected in the East China Sea in a depth of 1110m was investigated. The oxygen isotopic composition and Mg/Ca ratios determined along center-to-surface segments are used as geochemical proxies for the assessment of seawater paleotemperatures. Calculations are based on the assumption that the calculated temperature near the surface of the spicule is identical with the average ambient temperature of 4°C. A seawater temperature of 1.9°C is inferred for the beginning of the lifespan of the Monorhaphis specimen. The temperature increases smoothly to 2.3°C, to be followed by sharply increased and variable temperatures up to 6–10°C. In the outer part of the spicule, the inferred seawater temperature is about 4°C. The lifespan of the spicule can be estimated to 11,000±3000years using the long-term trend of the inferred temperatures fitted to the seawater temperature–age relationships since the Last Glacial Maximum. Specimens of Monorhaphis therefore represents one the oldest living animals on Earth. The remarkable temperature spikes of the ambient seawater occurring 9500–3100yearsB.P. are explained by discharges of hydrothermal fluids in the neighborhood of the spicule. The irregular lamellar organization of the spicule and the elevated Mn concentrations during the high-temperature growth are consistent with a hydrothermal fluid input.

Fibrous gels of cetylpyridinium chloride in binary solvent mixtures: structural characteristics and phase behaviour

Molecular gels of cetylpyridinium chloride (CPC) in binary solvent mixtures have been investigated. CPC formed a turbid gel at a critical solvent composition of 3 : 1 v/v CHCl3 : H2O. The gelation ability was evaluated in various other organic solvents in the presence of water and the phase evolution was studied. The microstructure of the CPC gel was proposed based on spectroscopic, microscopic and small-angle X-ray scattering (SAXS) techniques. A detailed molecular level investigation using SAXS and molecular modelling demonstrated the gel to assemble as a lamellar organization maintaining a highly interdigitated bilayer structure with CPC molecules. Concentration dependent hierarchical self-assembly from micelles to fibrous gels was thus elucidated with highly interdigitated lamellar gel organization resulting from intense hydrophobic interactions in the tail-to-tail arranged bilayers. The stimuli responsiveness of the gel was elucidated with pH dependent entrapment of Congo Red.