Chirality effects at each amino acid position on tripeptide self-assembly into hydrogel biomaterials.

Hydrogels formed by ultrashort peptides are emerging as cost-effective materials for cell culture. However, L-peptides are labile to proteases, while their D-isomers are thought to not support cell growth as well. In contrast, the self-assembly behaviour and biological performance of heterochiral peptides (i.e., made of both d and l amino acids) are largely unknown. In this study, we evaluate the effects of amino acid chirality on tripeptide self-assembly and hydrogelation at physiological pH, and cytocompatibility in fibroblast cell culture. A series of uncapped hydrophobic tripeptides with all combinations of d, l amino acids was prepared, tested for self-assembly under physiological conditions, and analysed by circular dichroism, FT-IR, cryo-TEM, AFM, and Thioflavin T fluorescence imaging. Amino acid chirality has a profound effect on the peptides' supramolecular behaviour. Only selected isomers form hydrogels, and of amyloid structure, as confirmed by rheology and XRD. Importantly, they are able to maintain the viability and proliferation of fibroblasts in vitro. This study identifies two heterochiral gels that perform well in cell culture and will assist in the design of innovative and cost-effective peptide gel biomaterials.

Synthesis and mesophases of glycerate surfactants.

In the quest to rationally design novel mesophase systems the challenge remains to deconvolute the relationship between structure, composition, and function. In the current study, novel glycerol-derived surfactants with high negative interfacial areas and a preference for inverse phase behavior have been targeted and synthesized. This has been achieved by application of the rule-of-thumb afforded by the critical packing parameter (CPP), namely, that inverse phase behavior is favored by wedge-shaped molecules with relatively small head group versus chain volume. Highly splayed hydrophobes with exaggerated cross section such as oleyl (cis-octadec-9-enyl) hexahydrofarnesyl (3,7,11-trimethyl-dodecyl) and phytanyl (3,7,11,15-tetramethyl-hexadecyl) were particularly successful for this purpose across many variations of head group. The phase behavior of the binary system in water of many of these surfactants is relatively simple. Typically, cubic or inverse hexagonal phases exist at the interface with water with the inverse micellar phases present at lower hydration. The inverse liquid crystalline phases were present for a broad range of temperatures and compositions.

Ricin antitoxins based on lyotropic mesophases containing galactose amphiphiles.

Lyotropic mesophases of the inverse hexagonal or cubic type are nanostructured materials that result from the self-assembly of amphiphilic surfactant molecules in water. The extremely large area of the surfactant-water interface inherent within these structures makes them attractive media for sorbent or encapsulant systems. Here, we report on the development of a new class of polyvalent materials that are based on the incorporation of bioactive ligands within lyotropic mesophases. In particular, we have studied the potential for these materials to behave as polyvalent antitoxins by incorporating synthetic galactose amphiphiles, which mimic the natural cell surface ligand for the protein toxin ricin. The study demonstrates that cubic morphology lyotropic mesophases containing galactose amphiphiles exhibit high specificity ricin uptake, with favorably high dissociation constants and high capacities. We suggest that lyotropic mesophase polyvalent ligands are thus promising materials for the incorporation of a broad range of cell surface recognition moieties and hence may have wide applicability as materials capable of partaking in biological recognition processes.

Monovalent and polyvalent carbohydrate inhibitors of ricin binding to a model of the cell-surface receptor.

A selection of galactose and lactose analogues was evaluated for their potency in inhibiting the binding of ricin to immobilised asialofetuin, which is a model of the cell-surface receptor for ricin. The aim was to identify compounds that could be used as antagonists of ricin toxicity in vivo, and as more selective, and therefore safer, antitoxins. Although one of these analogues had been identified by molecular modelling in a previous study as a potentially potent inhibitor, it and the other carbohydrates studied were less effective than galactose and lactose themselves (I(50) = 1.39 and 0.74 mM, respectively). In an attempt to increase the potency of carbohydrate-based inhibitors, galactose was coupled to the surface of dendrimers. No synergistic interactions were observed from this multivalent approach. Encouraging results, however, were obtained with a self-assembled lyotropic mesophase gel containing novel synthetic galactose-based surfactants, which was able to sequester ricin from aqueous solution in a 2-phase system.

Application of evanescent wave optics to the determination of absolute distance in surface force measurements using the atomic force microscope.

A combined scanning near field optical/atomic force microscope (AFM) is used to obtain surface force measurements between a near field sensing tip and a tapered optical fibre surface, whilst simultaneously detecting the intensity of the evanescent field emanating from the fibre. The tapered optical fibre acts as a compliant sample to demonstrate the possible use of the near field intensity measurement system in determining 'real' surface separations from normal AFM surface force measurements at sub-nanometer resolution between deformable surfaces.

The influence of the modification of etched bovine dentin on bond strengths.

OBJECTIVE: The aim of this study was to modify demineralized bovine dentin surfaces by application of either 12.5% sodium hypochlorite (NaOCl), or 0.1% (w/w) Type I collagenase, after conditioning with phosphoric acid, to observe the demineralized surface and to investigate the effect on tensile bond strength. METHODS: The NaOCl was applied to etched dentin for 30 s, 1 or 2 min and the collagenase for 1, 3 or 6 h. A control group was used without NaOCl or collagenase treatment. Prior to bonding, treated surfaces were examined using an Atomic Force Microscope (AFM). A 2.3 mm diameter area of dentin was conditioned, treated and bonded with either One Coat Bond or Single Bond following each manufacturer's instructions, and a resin composite rod attached. Bonds were stressed in tension at a rate of 1 mm/min until failure. Mean bond strengths were calculated (MPa) and mode of failure was determined by observation at 20x magnification. Results were analyzed using multiple regression analysis and LSD test at the 95% level of confidence (n = 12). RESULTS: AFM results showed progressive changes of the surface collagen as the treatment time of NaOCl or collagenase increased. For both bonding systems, the bond strengths of 1 min NaOCl and 3 h collagenase treatments were significantly higher than the control or other treatment groups (p < 0.05). Bond failure consisted of mostly adhesive failure between dentin and resin combined with small regions exhibiting cohesive failure of resin. SIGNIFICANCE: Bond strengths were not dependent on the thickness of the hybrid layer, but rather quality of the hybrid layer.

Computer recognition of regional lung disease patterns.

We have developed an objective, reproducible, and automated means for the regional evaluation of the pulmonary parenchyma from computed tomography (CT) scans. This method, known as the Adaptive Multiple Feature Method (AMFM) assesses as many as 22 independent texture features in order to classify a tissue pattern. In this study, the six tissue patterns characterized were: honeycombing, ground glass, bronchovascular, nodular, emphysemalike, and normal. The lung slices were evaluated regionally using 31 x 31 pixel regions of interest. In each region of interest, an optimal subset of texture features was evaluated to determine which of the six patterns the region could be characterized as. The computer output was validated against experienced observers in three settings. In the first two readings, when the observers were blinded to the primary diagnosis of the subject, the average computer versus observer agreement was 44.4 +/- 8.7% and 47.3 +/- 9.0%, respectively. The average interobserver agreement for the same two readings were 48.8 +/- 9.1% and 52.2 +/- 10.0%, respectively. In the third reading, when the observers were provided the primary diagnosis, the average computer versus observer agreement was 51.7 +/- 2.9% where as the average interobserver agreement was 53.9 +/- 6.2%. The kappa statistic of agreement between the regions, for which the majority of the observers agreed on a pattern type, versus the computer was found to be 0.62. For regional tissue characterization, the AMFM is 100% reproducible and performs as well as experienced human observers who have been told the patient diagnosis.

Quantification of idiopathic pulmonary fibrosis using computed tomography and histology.

We used computed tomography (CT) and histologic analysis to quantify lung structure in idiopathic pulmonary fibrosis (IPF). CT scans were obtained from IPF and control patients and lung volumes were estimated from measurements of voxel size, and X-ray attenuation values of each voxel. Quantitative estimates of lung structure were obtained from biopsies obtained from diseased and normal CT regions using stereologic methods. CT density was used to calculate the proportion of tissue and air, and this value was used to correct the biopsy specimens to the level of inflation during the CT scan. The data show that IPF is associated with a reduction in airspace volume with no change in tissue volume or weight compared with control lungs. Lung surface area decreased two-thirds (p < 0.001) and mean parenchymal thickness increased tenfold (p < 0.001). An exudate of fluid and cells was present in the airspace of the diseased lung regions and the number of inflammatory cells, collagen, and proteoglycans was increased per 100 g of tissue in IPF. We conclude that IPF reorganized lung tissue content causing a loss of airspace and surface area without increasing the total lung tissue.

High-resolution CT-derived measures of lung density are valid indexes of interstitial lung disease.

To assess the validity of computer-assisted methods in analyzing the lung parenchyma imaged with high-resolution computed tomography (HRCT), we compared computer-derived estimates of lung density to other, more traditional, measures of parenchymal injury in 24 subjects with idiopathic pulmonary fibrosis (IPF) and 60 subjects with extensive occupational exposure to asbestos. Gray scale density histograms were constructed from the HRCT images. The gray scale histogram of both study groups was of a skewed unimodal distribution. However, compared with the asbestos-exposed subjects, the patients with IPF had a gray scale distribution that was significantly shifted to the right (greater density) and flatter. In a multivariate analysis, after controlling for age and cigarette smoking, we found that the mean and median gray scale densities were independently associated with the presence of moderate-to-severe dyspnea, a higher International Labour Office chest X-ray category, a lower forced vital capacity, and a higher concentration of macrophages and eosinophils in the bronchoalveolar lavage fluid. These factors accounted for > 70% of the variance of the mean and median gray scale densities. Interestingly, no differences in gray scale density measures were noted between patients with IPF and patients with asbestosis when these other factors were taken into account. Our results suggest that computer-derived density analysis of the lung parenchyma on the HRCT scan is a valid, clinically meaningful, and objective measure of interstitial lung disease.